Cyanobacteria are known for their extensive and highly visible blooms in rivers or dams in Africa. One of the most important cyanobacteria is Microcystis aeruginosa which can synthesise various microcystins that may affect the health of humans and animals. Accurate and efficient detection of microcystins in water is thus important for public and veterinary health. Two enzyme-linked immunosorbent assays (ELISA), a commercially-available ELISA kit (Abraxis) and a newly-developed Norwegian ELISA (putatively cheaper and more robust) were used to detect microcystins in fresh water in South Africa. Water samples were collected monthly at two sites, the Hartbeespoort Dam and a crocodile breeding dam. Extremely high microcystin concentrations (exceeding 360 μg L−1) were detected in the Hartbeespoort Dam during January 2015, whereas the microcystin concentrations in the crocodile breeding dam peaked during March–April 2015. Both ELISAs were positively correlated when analysing water samples ‘as is’ and following resin adsorption and methanol extraction. However, following resin adsorption and methanol extraction of the water samples, the correlation between the two assays was much stronger. These results suggests that the two ELISAs provide comparable results. If the Norwegian-developed ELISA can be packaged and made available as a user-friendly kit, it could be used successfully in surveillance programmes to monitor microcystin concentrations in fresh water bodies in Africa.
The cytotoxicity of three Stenocarpella maydis metabolites (diplodiatoxin, dipmatol and diplonine) was investigated on Neuro-2a, CHO-K1 and MDBK cell lines. Diplodiatoxin was the most cytotoxic followed by dipmatol. Conversely, diplonine was not cytotoxic.Diplodiatoxin and dipmatol affected mitochondrial succinate dehydrogenase (MTT assay) and the overall viability of cells as assessed in real-time (xCELLigence assay). The results obtained so far indicate that diplodiatoxin and dipmatol exert their toxicity possibly via the necrotic cell death pathway.
Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in ‘vermeersiekte’ and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.
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