Blooms of cyanobacteria have increased in occurrence in the past three decades and have been reported to cause severe problems for animals and humans, leading to death in extreme instances. The majority of poisonings that have taken place have been attributed to a hepatotoxin produced by the species Microcystis aeruginosa, namely microcystin. The appearance of a cyanobacterial bloom does not give any indication as to its toxicity and therefore, it is imperative that simple, yet sensitive, bioassays are developed to overcome this problem. This study was undertaken to evaluate the effects of microcystin-LR on the liver of Xenopus laevis both in vitro and in vivo. This animal provides an opportunity to study the long-term hepatotoxic effects of the toxin compared to in vitro studies performed with mice and rats. The use of the liver slice model system as a potential bioassay to study the effects of microcystin-LR on Xenopus laevis liver was evaluated. Liver slices were cultured in RPMI- 1640 culture medium for periods ranging from 30 hours to 10 days and the liver slices were exposed to toxin concentrations ranging from 1nM to 500nM. The use of frog liver slices to study the longer-term effects of low-dose exposure to microcystin-LR was evaluated by observing the ultrastructural changes within hepatocytes using transmission electron microscopy, the release of the enzymes alanine aminotransferase and lactate dehydrogenase into the surrounding culture medium, as well as using a 3-[4,5-dimethylthiazol-2yl]-2,5- diphenyl tetrazolium bromide assay to determine the viability of the liver slices in culture. The amount of lipid peroxidation in the liver slices after exposure to microcystin-LR was assessed using the Thiobarbituric Acid Test. Results showed the frog liver slice culture system to be an inadequate method to evaluate the hepatotoxic effects of microcystin-LR. An in vivo assessment of the effects of microcystin-LR on Xenopus laevis was carried out using a total of 9 frogs (3 groups of 3 frogs). Frogs received a single intraperitoneal dose of 120mg/kg of microcystin-LR and were sacrificed at 8 and 24 hours post exposure. Microcystin-LR caused no significant change in serum lactate dehydrogenase levels, hepatosomatic index (liver weight as a percentage body weight), glutathione peroxidase activity, glycogen or lipid peroxidation. There was, however, an increase in glutathione sii transferase activity in the liver. The presence of the toxin in the liver was confirmed by immunohistochemistry. This study suggests that Xenopus laevis has, in some way, adapted to detoxifying aquatic toxins in the environment.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:nmmu/vital:11069 |
| Date | January 2003 |
| Creators | Coates, Nadya |
| Publisher | University of Port Elizabeth, Faculty of Science |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis, Masters, MSc |
| Format | xviii, 131 leaves, pdf |
| Rights | Nelson Mandela Metropolitan University |
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