Abstract
M.Sc.
A lot of research in ecotoxicology is currently focused on identifying and developing suitable biomarkers for use in assessing the health of aquatic systems. Thus this study was done as part of a major project aimed at identifying and selecting suitable biomarkers to use in the Rietvlei System, South Africa. This study focused on the response of Clarias gariepinus (Burchell, 1822) to contaminants using three biomarkers: population genetic structure, DNA damage and metallothioneins (MT¡¦s). The main objectives of the study were to assess the potential of these biomarkers in C. gariepinus and to provide baseline data on the Rietvlei System. Eighteen enzyme loci (creatine kinase (Ck), esterase (Est), phosphoglucomutase (Pgm), glycerol-3-phosphate dehydrogenase (G3pdh-1 and -2), glucose-6-phosphatase isomerase (Gpi-A and -B), L-lactate dehydrogenase (Ldh-A and B), malate dehydrogenase (sMdh-1, -2 and -3), malate dehydrogenase (NADP+) (Mdhp), phosphogluconate dehydrogenase (Pgdh), isocitrate dehydrogenase (NADP+) (Idh); superoxide dismutase (Sod), and peptidase: substrate (leucyl tryrosine) (Pep-S1 and -2)) were screened for by horizontal starch gel electrophoresis. An agarose gel electrophoretic method was used for DNA damage determination and a spectrophotometric method for metallothionein. The Rietvlei System population (RSP) showed higher genetic variability as shown by the average number of alleles per locus (A), proportion of polymorphic loci (P) and expected average heterozygosity (HE) than the reference populations. Statistically significant deviations (p < 0.05) from Hardy-Weinberg proportions associated with heterozygote deficiencies occurred at Gpi-B, Idh, Pgdh, Ldh-A and Pep-S1 for RSP. These deviations could have been a result of small sample size. There was high genetic differentiation between RSP and the reference populations with FST values of 0.450 and 0.480 between RSP and wild population (WP) and RSP and Northern Cape population (NCP) respectively. A mean base-pair length of 6770.9 „b 10.68 (SE) bp was recorded in the Marais Dam population (MDP) that of 6327.2 „b 17.69 (SE) bp was recorded in the Rietvlei Dam population (RDP). The MDP had a higher proportion of DNA fragments with low base-pair lengths than RP indicating a higher level of DNA damage. The level of DNA damage in the MP was higher than in the RDP. The mean MT¡¦s content was 3.63 „b 0.643 (SE) nmol g-1 wet weight in the MDP and 4.35 „b 0.580 (SE) nmol g-1 wet weight in the RDP. There was no significant (p = 0.40) difference in the levels of MT¡¦s between the two dams. It was concluded that there were significant differences in genetic diversity between RSP and the other populations in unpolluted sites; hence genetic diversity in C. gariepinus has potential as biomarker of pollution. However, further research is recommended to investigate possible contributing factors to differences in genetic diversity such as inter-sex males, geographical isolation and differences in sample sizes used. DNA damage is also a potential biomarker provided due care is taken in interpreting average base-pair length data to consider differences in each DNA size class. It seems further research is required before conclusions are made about use of MT¡¦s as biomarker in this population. The levels of MT¡¦s in other uncontaminated systems will be useful to determine whether the levels recorded in this study are background levels or responses to contamination.
Prof. F.H. van der Bank