The content of zearalenone and its metabolites in urine and tissue samples from pigs fed zearalenone-contaminated oats was established by analytical methods combining solid-phase extraction cleanup of the samples with highly selective liquid chromatography-mass spectrometry (LC-MS)/MS detection. Investigation of the urine samples revealed that approximately 60% of zearalenone was transformed in vivo to alpha-zearalenol and its epimer beta-zearalenol in a mean ratio of 3:1. Zeranol and taleranol as further metabolites could only be detected in trace amounts. Zearalanone was identified at considerable concentrations, though only in a couple of samples. In contrast, liver samples contained predominantly alpha-zearalenol, and to a minor extent beta-zearalenol and zearalenone, with a mean ratio of alpha-/beta-zearalenol of 2.5:1, while zeranol, taleranol, or zearalanone could not be identified in any of the investigated samples. The degree of glucoronidation was established for zearalenone as 27% in urine and 62% in liver; for alpha-zearalenol as 88% in urine and 77% in liver; and for beta-zearalenol as 94% in urine and 29% in liver. Analyses of muscle tissue revealed relatively high amounts of nonglucuronidated zeranol and alpha-zearalenol together with traces of taleranol and zearalenone, indicating that the metabolism of zearalenone and its metabolites is not restricted to hepatic and gastrointestinal metabolic pathways.
A group of five heifers were fed for 84 days with 2 kg of zearalenone-contaminated oats (1370 microg/kg) resulting in an average daily intake of 2740 microg of zearalenone per animal. In a parallel experiment five heifers were implanted with two 25 mg zeranol pellets, one at the beginning of the study and one after 42 days, and fed with 2 kg of "blank" control oats (79 microg/kg, daily intake = 158 microg). A third group of five animals were also fed with 2 kg of "blank" oats and served as control. Urine samples of all animals were collected every 5-6 days during the whole period of the study. Animals of all three groups were killed 84 days after the beginning of the feeding study. Tissue samples (back, femoral region, liver, and residues of implanted pellets) were taken during post-mortem investigations. The content of zearalenone and zeranol and their metabolites in urine and tissue samples was established by an analytical method combining solid-phase extraction and high-performance liquid chromatography-tandem mass spectrometry. Urinary excretion rates of zeralenone and zeranol were calculated from these results.
Two trials were carried out to compare the use of different enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against Sarcoptes scabiei var. suis. In the first trial, we employed four tests and 70 selected sera from a closed pig farm with sarcoptic mange. The sera were taken from the breeding area as well as from the finishing unit and compared with skin scrapings. The SARCOPTES-ELISA 2001 yielded the most positive results (88.58%), followed by the ELISA of the National Veterinary Institute, Uppsala (70%), the Acar-Test P-ELISA (52.86%), skin scrapings (48.57%), and the CHEKIT Sarcoptest (30%). In the second trial, eight litters from infected sows were examined from weeks 1-12 of life using the CHEKIT Sarcoptest and the SARCOPTES-ELISA 2001. The presence of maternal antibodies was highest on day 7 in both ELISAs and could be detected until weeks 5-9 of life. Antibodies increased as a result of an active immune response between days 56 and 63 in the SARCOPTES-ELISA 2001 and between days 70 and 77 of life in the CHEKIT Sarcoptest. Significant differences between the first and second litters were observed.
An understanding of the conditions influencing protein binding of catecholamines (CAs) is important in studying their metabolic effects. Unfortunately, reports on plasma protein binding of CAs are scarce, conflicting and mainly performed in vitro. The aim of our in vivo and in vitro studies was to investigate binding and clearance of radioactive adrenaline (epinephrine) ((3)H-A), noradrenaline (norepinephrine) ((3)H-NA) and their metabolites in sheep blood. The time course of the radioactivity in the blood after intravenous injection of (3)H-A and (3)H-NA (3.7 MBq each) in 4 sheep (2 of each sex; total of 8 administrations) was determined. Blood samples were taken from the jugular vein. The highest radioactivity was observed in the first sample (5 min) following injection. Radioactivity showed a biphasic disappearance. An initial stage, in which radioactivity decreased rapidly (within 1 h) after the injection, was followed by a slow stage, lasting for up to 1 month, until background levels were reached. In vitro results indicated that NA and A were present not only in plasma (70%) but also in the erythrocytes (30%; mainly bound to haemoglobin). Sephadex G-25 gel filtration revealed that from the plasma fraction about 15% was strongly bound to proteins (mainly albumin). These results demonstrate that previous experiments in this field have overestimated the percentage of CAs bound to plasma proteins, because binding to haemoglobin was previously not known. In the future, efforts should be made to characterize the adduct products of CAs and establish an assay to determine them in vivo. If this could be achieved, it would yield a valuable tool for measuring the stress experienced for a longer period.
This paper describes a feeding study with 7 pigs, which were fed with deoxynivalenol contaminated oats at a level of 0.23 mg/kg body mass/day over 16 experiment days. The contamination level of consumed feed was 14.4 mg DON/kg in the ration. The parallel control group of 7 pigs were fed with DON free oats. Urine samples were taken each two days. The content of DON and DOM-1 (de-epoxy deoxynivalenol) in urine was determined. The mean concentration of DON in urine of animals in trial group was 580 μg/l, whereas DOM-1 32 μg/l.ZUSAMMENFASSUNG: In einer Fütterungsstudie wurden Schweine in zwei Gruppen zu je 7 Tieren mit kontaminiertem (Versuchsgruppe) und unbelastetem Getreide (Kontrollgruppe) gefüttert. Die Gesamtkonzentration von DON in der Ration der Versuchsgruppe war 14.4 mg/kg. Die Tagesdosis betrug in der Versuchsgruppe 0.23 mg/kg Körpermasse und Tag. Der Versuch dauerte 16 Tage, jeden zweiten Tag wurde der Spontanharn gesammelt. Die Urinproben wurden auf DON und DOM-1 analysiert. Für die Analyse der Urinproben bzw. des Futtermittels wurde eine HPLC-MS Methode verwendet.
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