The effect of tilmicosin on cardiac superoxide dismutase and glutathione peroxidase activities

In this study, the effect of tilmicosin on cardiac superoxide dismutase and glutathione peroxidase activities was investigated. Forty male BALB/c mice were used as material. Ten mice served as a control group, and 30 mice were injected with tilmicosin (25 mg/kg body weight, subcutaneously, with a single injection). After drug administration, they were monitored for 3 days. Tilmicosin caused decreases in cardiac superoxide dismutase and glutathione peroxidase activities.     

Detection of superoxide anion generation by equine spermatozoa

OBJECTIVE: To identify the generation of the superoxide anion by equine spermatozoa. SAMPLE POPULATION: Multiple ejaculates collected from 3 Thoroughbred stallions. PROCEDURES: Induced superoxide production by reduced nicotinamide adenine dinucleotides (NAD[P]H; ie, reduced nicotinamide adenine dinucleotide [NADH] and reduced nicotinamide adenine dinucleotide phosphate [NADPH]) was measured by use of a nitroblue tetrazolium (NBT) reduction assay on whole spermatozoa and a cytochrome c reduction assay on isolated membrane fractions of spermatozoa. Localization of superoxide generation was determined by use of NBT cytochemistry. RESULTS: A dose-dependent increase in NBT reduction was found in the presence of NADPH, which was inhibited by superoxide dismutase (SOD). The flavoprotein inhibitor, diphenyleneiodonium (DPI; 5 or 15 microM), significantly decreased NBT reduction. Cytochrome c reduction by plasma membranes of spermatozoa was significantly higher in the presence of NADPH than in its absence. Cytochemical staining of equine spermatozoa in the presence of NADPH and NADH revealed diaphorase labeling in the spermatozoon midpiece and head. This staining was inhibited by DPI and SOD. CONCLUSIONS AND CLINICAL RELEVANCE: Results of our study indicate that superoxide generation is associated with a membrane-associated NAD(P)H oxidase present in equine spermatozoa, although mitochondrial generation of superoxide is also detected. This oxidase may play a role in cell signaling or may also contribute to cytopathic effects associated with oxidative stress in equine spermatozoa.     

Purification and quantification of lactoferrin in equine seminal plasma

Lactoferrin with a molecular mass of 80 kDa was purified from equine seminal plasma by heparin-Agarose affinity chromatography and Sephacryl S-200 gel filtration. Purified lactoferrin was found to be highly homogeneous on the bases of its migration as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and of the monospecificity of rabbit antibodies to the purified protein in immunoblotting of seminal plasma proteins. A sandwich enzyme-linked immunosorbent assay was developed for quantifying lactoferrin in equine seminal plasma. Seminal plasma lactoferrin concentrations in 23 normal stallions ranged from 42 to 453 microg/ml, with a mean value of 157 +/- 118 microg/ml (S.D.).     

Level of superoxide dismutase,glutathione peroxidase and uric acid in thioacetamide-induced cirrhotic rats

Levels of superoxide dismutase and glutathione peroxidase were determined in blood and hepatic tissues of thioacetamide-induced cirrhotic rats and compared to levels in age-matched control animals. The plasma level of uric acid was also determined in these animals. A general decrease was noticed in the level of all the antioxidants examined as compared to the control. This decrease was statistically significant in the level of all the antioxidants studied, except for the level of superoxide dismutase in blood. A decrease in the antioxidant level may indicate an increase in free radical level and thereby an increase in cellular damage in cirrhotic rats. The changes in the level of antioxidants showed a direct correlation with the changes in the level of trace elements observed in our previous studies. These studies suggest that antioxidants alone or in combination with trace elements may have beneficial effects in treating liver cirrhosis.     

Pharmacokinetics of tilmicosin in equine tissues and plasma

The macrolide antibiotic tilmicosin has potential for treating bacterial respiratory tract infections in horses. A pharmacokinetic study evaluated the disposition of tilmicosin in the horse after oral (4 mg/kg) or subcutaneous (s.c.) (10 mg/kg) administration. Tilmicosin was not detected in equine plasma or tissues after oral administration at this dose. With s.c. injection, tilmicosin concentrations reached a maximum concentration of approximately 200 ng/mL in the plasma of the horses. Tilmicosin concentrations in plasma persisted with a mean residence time (MRT) of 19 h. Maximum tissue residue concentrations (C(max)) of tilmicosin measured in equine lung, kidney, liver and muscle tissues after s.c. administration were 2784, 4877, 1398, and 881 ng/g, respectively. The MRT of tilmicosin in these tissues was approximately 27 h. Subcutaneous administration of tilmicosin resulted in severe reactions at the injection sites.     

Activity of superoxide dysmutase, catalase and glutathione peroxidase in rats exposed to chlorpyrifos and erofloxacin

The aim of the study was to investigate the influence of administration of chlorpyrifos and/or enrofloxacin on the activity of chosen antioxidative enzymes i.e.: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in erythrocytes of rats. Chlorpyrifos was administered by stomach tube during 28 days at a dose of 3 mg/kg bw (0.02 LD50), and enrofloxacin was administered by stomach tube at a dose of 5 mg/kg bw during 3 subsequent days. It was stated that administration of enrofloxacin at applied dose did not cause any major changes in the activity of investigated antioxidative enzymes. The four-week exposure of rats to chlorpyrifos caused noticeable decrease in SOD and CAT activity in erythrocytes of rats at the beginning of the experiment (up to 24th hour) in comparison with the control group. The activity of GPx during all periods of the experiment was increased. In the group of animals in which both chlorpyrifos and enrofloxacin were applied, the profile of changes in activity of examined enzymes was similar to that one, which was observed after administration of chlorpyrifos exclusively, what may indicate lack of co-action between compounds used in the experiment.     

Quantification of plasma reduced glutathione, oxidized glutathione and plasma total glutathione in healthy cats

Glutathione is an important intracellular tripeptide with multiple functions. Abnormal glutathione metabolism is thought to play an important role in various diseases of cats. However, no data regarding concentration of plasma glutathione are available for domestic cats. This study discusses the development of a rapid, simple high pressure liquid chromatography method for measurement of reduced glutathione (GSH), oxidized glutathione (GSSH) and total glutathione (GSHt) in plasma, for the purpose of establishing baseline data for future studies. The mean concentrations of GSH, GSSH and GSHt were 4.51+/-1 microM; 19.44+/-3.79 microM (expressed as GSH equivalent) and 23.59+/-3.89 microM, respectively. This is the first report of plasma glutathione concentrations in this species.     

The stability of glutathione peroxidase activity in plasma from cattle,pigs and sheep on storage in the presence and absence of glutathione

Ovine, bovine and porcine plasma glutathione peroxidase (GSH-Px) activity decreased on storage at both 4°C and 20°C. The ovine and bovine enzymes were significantly less stable than the porcine enzyme. The addition of GSH to a final concentration of 2 mmol/L to plasma samples at the commencement of storage retarded the loss of both ovine and bovine plasma GSH-Px activity. The ovine enzyme was unique in that after inactivation by storage at 4°C, incubation with GSH restored the enzyme activity. It is recommended that plasma GSH-Px should be assayed fresh, or otherwise stored at –20°C.     

Effect of heterologous seminal plasma and semen extenders on motility of frozen-thawed ram spermatozoa

Ram seminal plasma increases the fertility of frozen-thawed ram spermatozoa deposited into the cervix. The aim of the current study was to compare the effect of ram seminal plasma to that of bull seminal plasma, dog prostatic fluid, protein-free TALP TrilEq (Triladyl with 0.5 mt of Equex STM paste added to each 100 mt) and heat-treated skim milk on longevity and percentages of progressively motile and aberrantly motile frozen-thawed ram spermatozoa. Three ejaculates from each of 6 rams were extended in TrilEq, pooled and frozen in straws as a single batch per ram. One hundred and eight straws (3 straws from each ram for each fluid) were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85 ml of the appropriate fluid at 37 degrees C and kept at that temperature for 6 h. Motility was assessed at x200 magnification immediately (time zero) and 2, 4 and 6 h after thawing. Progressive motility decreased from each time to the next (P < 0.05) and was 39.0 % (0 h), 26.0 % (2 h), 19.6 % (4 h) and 12.6 % (6 h); SEM 1.24, n = 108 for each group. Ram seminal plasma resulted in higher progressive motility than bull seminal plasma, lower than milk, and similar to the other fluids. Ram seminal plasma resulted in lower aberrant motility than protein-free TALP and similar aberrant motility to other fluids. The effect of ram seminal plasma and dog prostatic fluid was very similar. The effect of ram seminal plasma on the fertility of frozen-thawed ram spermatozoa deposited into the cervix is not due an exceptionally beneficial effect on the motility of spermatozoa.     

Laminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis

REASONS FOR STUDY: Xanthine oxidase (XO)-dependent production of superoxide anion and hydrogen peroxide, a characteristic of ischaemia-reperfusion injury, may contribute to the development of equine laminitis. OBJECTIVE: To determine the levels of XO and antioxidant enzymes (catalase, superoxide dismutase [SOD]) in the digital laminae of normal horses (CON) and horses in the developmental stage of laminitis using the black walnut extract (BWE) model. METHODS: Healthy horses (n = 12) were administered BWE (BWE group, n = 6), or water (CON group, n = 6) through a nasogastric tube. At the onset of leucopenia in the BWE-treated animals, all horses were anaesthetised, digital laminae and other samples collected rapidly and flash frozen, and the animals subjected to euthanasia. Extracts of the frozen tissues were assayed for the 2 conformational forms of xanthine: oxygen oxidoreductase (XOR), namely, xanthine dehydrogenase (XDH) and xanthine oxidase (XO), as well as the antioxidant enzymes, SOD and catalase. RESULTS: Extracts of liver, lungs and skin, but not digital laminae, from either CON or BWE-treated horses had endogenous SOD, whereas all had endogenous XO and catalase. The levels of XDH, XO and catalase were similar in extracts of laminae from CON and BWE-treated horses as was the ratio of XDH to XO in extracts. CONCLUSIONS AND POTENTIAL RELEVANCE: The absence of increased XO activity suggest against the involvement of this reactive oxygen intermediate-generating system in the development of laminar pathology in BWE-treated horses. Conversely, the absence of SOD from extracts of equine digital laminae, but not other tissues, suggests that the equine digital laminae are highly susceptible to damage by superoxide anion, produced, for example, by emigrant inflammatory leucocytes.     

Serum selenium concentrations and glutathione peroxidase activities in cattle grazing forages of various selenium concentrations

In cows from 15 dairy herds (n = 210), serum selenium (Se) concentrations ranged from 0.021 to 0.789 microgram/ml, whereas 0.05 to 0.40 microgram/ml is the reported range for adequate serum Se concentrations in cattle. Serum Se concentrations of dairy cattle appeared to follow a geographic distribution pattern. On the basis of herd mean serum Se concentrations, adequate serum Se concentrations were found in cattle from only 1 of 5 herds grazing forage in the geographic area classified as Se deficient for cattle. Adequate mean serum Se concentrations were found in cattle from 4 of 5 herds located in geographic areas described as having variable forage Se concentrations (Se-marginal areas). Of the 10 herds from these 2 areas, there were only 2 herds in which 95% of the cattle had serum Se concentrations in the Se-adequate range (0.05 to 0.40 microgram/ml). In 2 selected neighboring farms in the Se-deficient area, cattle in 1 herd had adequate serum Se concentrations and cattle in the other herd had less than adequate serum Se concentrations (less than 0.05 microgram/ml). Therefore, more cattle are at risk of developing Se-deficiency disease than is commonly believed and forage of neighboring farms may have different Se concentrations. Serum Se concentrations (up to 0.789 microgram/ml) correlated with glutathione peroxidase enzyme activity; this serum Se concentration (0.789 microgram/ml) is approximately 6.2 times higher than previously reported in dairy cattle. Therefore, RBC glutathione peroxidase activity may be useful in determining the diagnosis of chronic Se toxicosis.     

Studies on serum tocopherol, selenium levels and blood glutathione peroxidase activities in calves with white muscle disease

For the purpose of clarifying the cause of white muscle disease (WMD) in calves, tocopherol and selenium levels and blood glutathione peroxidase (GSH-Px) activity were measured on 10 calves with WMD and nine of their dams. The main clinical symptoms of the 10 calves with WMD were motor disturbances including recumbency and stiffness. Serum enzyme activities (GOT, GPT, CPK, LDH) in calves with WMD increased markedly, and this increase was also observed in some of their dams. Serum tocopherol levels of calves with WMD were low, 70% of which showing deficient levels of less than 70 micrograms/100 ml. Serum selenium levels of all the calves were lower than 35 ppb, indicating a deficiency, and were accompanied by low blood GSH-Px activity. alpha-Tocopherol and selenium concentrations in organs were very low. Dams of calves with WMD showed low serum tocopherol levels, 22% of which indicating deficient levels below 150 micrograms/100 ml. Serum selenium levels in dams showed a marked decrease to under 20 ppb, and also low blood GSH-Px activity. Feedstuffs supplied in the farms to affected calves indicated very low alpha-tocopherol contents (below 3 mg/100g DM) and low selenium concentrations below 50 ppb in DM. It was concluded that WMD in calves was attributable to nutritional muscular dystrophy caused by deficiencies in tocopherol and selenium in feedstuffs supplied to their dams.