A simple method for the determination of chloramphenicol in 22 matrices was prepared based on the QuEChERS and HPLC-MS/MS combination. Following a hydrolysis step, the homogenized samples were extracted and partitioned after adding sodium chloride with acetonitrile. Chloramphenicol was analysed by HPLC-MS/MS in negative electrospray mode by monitoring the daughter ions m/z: 321→194 and 321→152. The limit of decision (CCα) was calculated at the range of 0.10 μg kg−1 to 0.15 μg kg−1 and detection capability (CCβ) from 0.12 μg kg−1 to 0.18 μg kg−1. Validation results showed that this method is suitable for the determination and confirmation of chloramphenicol in various matrices.
Antioxidative/oxidative balance is crucial for proper functioning of cells and tissues. It is suggested that this balance can be partly controlled by sex steroid hormones and in consequence can exhibit age- and sex-related dependency. The aim of present study was to describe sex- and age-related changes in the activity of glutathione peroxidase (GSH-Px) with respect to total antioxidant activity (TAC) in reproductive organs of cattle. Biological samples were collected from slaughterhouse and comprised of ovaries, uterus, testes as well as livers as reference tissue. Animals were divided into group of bulls (aged between 13 and 24 months; n = 12), cows (aged between 14 and 27 months; n = 12) and female calves (aged between 2 weeks and 2 months; n = 12). Examined parameters were determined spectrophotometrically and the presence of GSH-Px isoform was confirmed by Western blotting technique. Activity of GSH-Px in genital tissues regardless of sex was significantly higher than in livers, while TAC showed opposite relationship. The differences in antioxidative parameters between testes and mature ovaries (e.g. GSH-Px-1.42 ± 0.47 nkat/mg prot vs. 1.08 ± 0.24 and 1.15 ± 0.23) were noticed as well as in chosen values between cows and female calves. Western blotting allowed the detection of cytosolic GSH-Px in all examined tissues with molecular weight around 21 kDa as monomer and around 84 kDa as tetramer depending on conditions of electrophoresis. The results may confirm the influence and regulatory role of sex steroid hormones on GSH-Px activity because the alterations were sex and age dependent.
The white-tailed eagle (Haliaeetus albicilla) is strictly protected in Poland due to its threat of extinction. This study’s main goal was to assess their exposure to indirect poisoning by anticoagulant rodenticides (AR). This study presents the investigation results of 40 white-tailed eagles’ suspected poisoning cases in the years 2018–2020 in Poland. In all tested liver samples, using a liquid chromatography–mass spectrometry method, at least one of the AR (bromadiolone, brodifacoum, difenacoum, flocoumafen) was detected and confirmed. The other tested AR compounds (chlorophacinone, coumachlor, coumatetralyl, difethialone, diphacinone, warfarin) were not detected. The mean concentration of the sum of rodenticides was 174.4 µg/kg (from 2.5 to 1225.0 µg/kg). In 20 cases, the sum concentration was above 100 µg/kg and in 10 cases it was above 200 µg/kg. Interpretation of cases of AR poisonings should take into account their concentration in the liver, anatomopathological lesions, circumstances of death/finding of the animal, and elimination of other possible causes of poisoning. Based on this study, AR was the direct cause of death in 10% of incidents. Extensive use of rodenticides generates a high risk of poisonings of white-tailed eagles in Poland.
IntroductionThe major difficulty in analysis of nitrofurans in feed, feed water, and food of animal origin is that nitrofurans have low molecular weights and fast metabolism. The principal goal of this study was to prepare a procedure for the determination of nitrofurans and their metabolites by a single method in different types of feed, feed water, and food of animal origin.Material and MethodsTwo-gram samples were subjected to hydrolysis and derivatisation processes by addition of hydrochloric acid and 2-nitrobenzaldehyde. After incubation the sample was purified by solid phase extraction technique. Nitrofurans were analysed using ultra-high-pressure liquid chromatography-MS/MS (UHPLC-MS/MS).ResultsThe results of validation fulfil the requirement of the confirmatory criteria according to the European Commission Decision 2002/657/EC regarding apparent recoveries (88.9%–107.3%), repeatability (2.9%–9.4%) and within-laboratory reproducibility (4.4%–10.7%).ConclusionThe method can be successfully applied to monitor nitrofurans and their metabolites in different matrices.
Introduction The study measured the hormonal and protein markers of acute stress, those of oxidative stress and total antioxidant capacity (TAC) in swine oral fluid, determined which of these parameters would be the most appropriate for future livestock welfare assessment and established the time when the samples should be taken. Material and Methods Stress was induced in 7 out of 14 castrated six-week-old Danbred×Duroc pigs by immobilisation on a nasal snare at 8 a.m., 1 p.m., and 6 p.m. and samples were taken both directly after the stressor was applied and 30 min later. The remaining pigs were the control group, which were not immobilised; their samples were taken at the same times. The concentrations of hormones and malondialdehyde (MDA) were measured using liquid chromatography with tandem mass spectrometry, while those of alpha-amylase and TAC were measured using spectrophotometry. Results The levels of cortisol and cortisone increased with statistical significance immediately after the acute stress response and 30 min later. A cut-off value set at 0.25 ng/mL cortisol concentration was capable of distinguishing between the stressed and control groups with 100% accuracy in evening samples and 95% accuracy overall. Prednisolone was not present, and the levels of testosterone and corticosterone were low and not distinctive. Alpha-amylase became significantly more concentrated during stress induction and 30 min later. The TAC and MDA levels rose after the stress but without statistical significance. Conclusion The most suitable markers of acute stress were cortisol, cortisone and alpha-amylase. Oral fluid is a reliable material for monitoring the level of pigs’ stress and should be collected in the evening.
ABSTRACT:The influence of age, gender, and type of tissue on superoxide dismutase (SOD) activity in bovine organs and tissues was investigated. The investigated material consisted of fragments of tissues and organs (liver, heart, lung, kidney, skeletal muscles, and diaphragm) from healthy cows (n = 15), bulls (n = 15), and female calves (n = 12) collected immediately after slaughter at the slaughterhouse. The total SOD activity was measured in tissue and organ homogenates by spectrophotometric method. PAGE electrophoresis and Western blotting technique with specific anti-SOD antibodies as well as zymography confirmed the presence of enzymatic protein and SOD isoenzymes in examined tissues. The study revealed significant differences in SOD activity between organs and tissues, which might be connected with different metabolic rate. Age-related changes were also observed. SOD activity was twice higher in kidneys, lungs, heart of calves compared to other groups of animals. It was also noticed that SOD activity was higher in younger animals and decreased with ageing among the group of bulls. Gender-related differences were observed in lungs and diaphragm. It was concluded that SOD activity depends on sex steroids, which is reflected in age and gender discrepancies, as well as metabolic rate of tissues.
The article reports a case of acute copper sulfate (CuSO4) poisoning in 2-day-old turkeys. The first incidences were noticed 2 h after placing the animals in the poultry house, where turkeys were provided with wood shaving bedding. Post-mortem examination showed wood shavings present in gastric and intestinal contents and numerous lesions in the alimentary tract: dark green color of the koilin lining, and surface erosions, deep ulceration, and severe congestion of the gizzard. The copper (Cu) concentration in samples of the liver, kidney, heart, gizzard content, gizzard muscle, gizzard lining, and wood shavings from the bedding was determined. Our results showed that the level of Cu in the livers of affected birds was more than sevenfold higher than the recommended value and the Cu content in bedding material was above the value considered to be toxic to turkeys. Our investigation identified the bedding disinfected using a CuSO4 solution as a possible cause of turkey fatalities.
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