Vermeersiekte or ‘vomiting disease’ is an economically important disease of ruminants following ingestion of Geigeria species in South Africa. Sheep are more susceptible and poisoning is characterised by stiffness, regurgitation, bloat, paresis and paralysis. Geigeria aspera was collected in the Vrede district (27° 25′ 48″ S; 29° 9′ 36″ E), Free State Province. The plant material was dried, milled and the toxic principles, known as sesquiterpene lactones, were extracted and isolated following chromatographic procedures. Even though geigerin and ivalin were previously isolated, an unknown sesquiterpene lactone, isogeigerin acetate, was also purified. Mouse myoblast (C2C12) and rat embryonic cardiac myocyte (H9c2) cell lines were exposed to different concentrations of geigerin, ivalin, isogeigerin acetate and a commercially available sesquiterpene lactone, parthenolide. An in vitro colorimetric assay 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used to assess cytotoxicity. The median effective concentrations (EC50) indicated that ivalin and parthenolide were significantly (p<0.05) more toxic than geigerin and isogeigerin acetate. A concentration-dependent cytotoxic response was observed in both cell lines, however, C2C12 cells were more sensitive. A high performance liquid chromatography (HPLC) analysis was used to evaluate the in vitro metabolism of parthenolide, following the addition of a mouse liver microsomal fraction. Results revealed that parthenolide, incubated with the microsomal fraction, undergoes enzymatic transformation to form a metabolite. / Dissertation (MSc (Veterinary Sciences))--University of Pretoria, 2020. / pt2021 / Paraclinical Sciences / MSc (Veterinary Sciences) / Unrestricted

Polychlorinated biphenyls (PCB) represent relevant persistent organopollutants of the environment and the estimated amount of PCB released into the environment is 750000 metric tons. White-rot fungi have been studied for long time due to their degradative potential toward various aromatic pollutants and it is known that these fungi are able to decompose PCB in vivo. Biodegradation of PCB by the fungus Pleurotus ostreatus was studied in the frame of this work. A high degradative efficiency of P. ostreatus was observed in the first set of experiments, even in the presence of relative high amount of added PCB. Fungus was able to transform 780±50 µg out of the intial amount 1000 µg in 20 ml of a cultivation media within 42 days. A decrease in toxicity was recorded during the degradation that suggests the suitability of this organism for a practical use in decontamination. In vitro experiments with purified laccase induced with Cu2+ from this fungus did not prove any participation of the enzyme in the first step of PCB transformation. The enzyme did not show an ability to degrade PCB even after purification from cultivation media containing PCB. It was found that the first step of PCB transformation can be performed by an intracellular process with microsomal fraction. A degradation of 44-67% was observed for...

A synthetic estrogen 17α-ethinylestradiol (EE2) is the main active component of the hormonal contraceptive pills. The rise of consumption of hormonal contraceptives has increased the risk of the back negative effects of EE2 to aquatic organisms. EE2 belongs to the endocrine disruptive compounds known for mimicking natural hormones. A more detailed examination of the transformation of this compound in vivo and in vitro can contribute to a better understanding of its negative effects. This master thesis is therefore devoted to the study of the metabolism of EE2 in two selected model organisms. The ligninolytic fungus Pleurotus ostreatus is the type of fungi with promising biodegradation ability to a lot of pollutants. These properties have led to numerous studies of the degradation potential of P. ostreatus towards EE2, with the possibility of removing this compound from the environment. EE2 has been degraded by the fungus P. ostreatus in vivo resulting in one hydroxylated metabolite, which estrogenic activity is in need for further study. In vitro studies were carried out with a microsomal fraction isolated from the mycelium of this fungus. The conversion of EE2 in vitro via CYPs dependent on NADPH has not been demonstrated, however using KHP as a cofactor, there was one metabolite of EE2 found,...