The conservation of biodiversity and endemism in South Africa’s freshwater aquatic ecosystems is a high priority, particularly in the Cape Floristic Region. However, the perception that South Africa lacks suitable fish species for recreational angling, aquaculture and biological control, led to the widespread introduction and use of alien fish species. As a result, formal stocking programs have seen the introduction of five of the world’s top 100 invasive species into South Africa (Dudgeon et al. 2006). According to Dudgeon et al. (2006) freshwater ecosystems are the most endangered ecosystem in the world. The threats to freshwater biodiversity, according to Dudgeon et al. (2006), can be grouped into five categories that interact with one another: overexploitation, water pollution, flow modifications, destruction of habitat and invasion by exotic species. This PhD study took place in the Amatola region of the Eastern Cape Province, South Africa. The Amatola region is a rural area with no large-scale mining or industrial developments, only localised settlements. These developments are mainly situated around impoundments, because of the resources such as water and food that they provide. Thus the dams within the Amatola region should theoretically have no major industrial stressors on them. The three impoundments studied were Binfield Park, Sandile and Wriggleswade Dams. Binfield Park Dam is a 260ha impoundment. It impounds the Tyume River and is used by both subsistence anglers from the local communities and occasionally by recreational bass anglers. Sandile Dam is a 146ha impoundment and is the smallest of the three dams in this study. It impounds the Wolf and Keiskamma Rivers. Wriggleswade Dam is a 1000ha impoundment used extensively by recreational bass and carp anglers, and impounds the Kubusi River. There is a paucity of information regarding the health of the indigenous and alien fish species from the study region, as well as on the parasite diversity of these various fish species. In order to fill the gaps in the information the following hypothesis was proposed. That the necropsy based and histology based fish health assessment can be successfully implemented as tools to assess the effects of heavy metal pollution and alien fish parasites in freshwater fish from selected impoundments in the Amatola region, Eastern Cape Province, South Africa. In order to achieve this hypothesis the main aim of this study will be to use the necropsy- and histology- based fish health assessment to determine the health status of the fish species in these impoundments as well as to understand the potential threat of water pollution and fish parasites. Fish were sampled with the aid of gill nets, fyke nets and by angling from each of the three impoundments over three surveys in July 2011, and March and August 2012. Following capture fish were transported to a field laboratory in aerated containers. At the field laboratory the fish were examined and dissected using the methods recommended by Adams et al. (1993) for a necropsy-based fish health assessment. Gills, livers, kidneys and gonads samples were also collected for histological analysis. Macroscopic and histology-based fish health assessment index was used, as well the analysis of muscle tissue of Micropterus salmoides and surface water and sediment from Binfield Park, Sandile and Wriggleswade Dams. It was shown that, according to the macroscopic fish health assessment index, M. salmoides in Wriggleswade Dam had a higher FHAI score compared to those in Binfield Park and Sandile Dam, there were no significant differences between the FHAI scores. However, the cause of the higher FHAI in the Wriggleswade Dam was because of the external skin damage caused by the presence of the alien parasite Lernaea cyprinacea. The histology-based fish health assessment index, however, showed that M. salmoides from Binfield Park had significantly higher histology Fish Index (IFISH) scores compared to those in Sandile and Wriggleswade Dams. The main contributors to the high IFISH score of Binfield Park were the significantly high Liver Index (IL) and Kidney Index (IK). The increased severity of the alterations observed in the liver and kidney tissue of the Binfield Park M. salmoides samples may have been as a result of the high concentration of mercury found in the muscle tissue of M. salmoides. The water quality and metals detected in the water of Binfield Park, Sandile and Wriggleswade Dams were all below the target water quality guideline values, as well as below those of previous research into the nutrients and presence of metals in these impoundments. The sediment metal analysis showed that the levels of Co, Mn and Ni were above the target guideline levels for Binfield Park, Sandile and Wriggleswade Dams, Cu was above target guidelines levels for Sandile Dam, and uranium was above the target guideline concentrations for Wriggleswade Dam. Binfield Park Dam had significantly high levels of mercury in the muscle tissue of M. salmoides, while Sandile Dam had significantly high levels of zinc in the muscle tissue of M. salmoides. It was shown that M. salmoides from each of the three impoundments are in a healthy state according to the parameters assessed. However, the presence of heavy metals, particularly mercury, uranium and zinc, do indicate the presence of human activities. The indigenous parasites of Anguilla mossambica have been well documented including the gastrointestinal nematode Paraquimperia africana, and the stomach nematode Heliconema africanum. Indigenous parasites such as the swimbladder nematode Anguillicola papernai had no effect on the condition factor of infected and uninfected eels. However, the damage caused by the alien parasites were evident, including the first documented effects of the alien gill monogenean Pseudodactylogyrus anguillae on indigenous wild populations of the longin fin eel A. mossambica from the Eastern Cape, South Africa. Histological observations indicated that an alien gill monogenean caused hyperplasia, increase in mucous cells, rupture of pillar cells as well as telangiectasia. This alien parasite has invaded the Keiskamma and Kei River systems in the Eastern Cape, South Africa. According to the macroscopic fish health assessment index, A. mossambica from Binfield Park, Sandile and Wriggleswade Dams are in a healthy state. However, the histology-based health assessment highlighted that the effects on P. anguillae have a severe negative impact on the health of A. mossambica. Using the macroscopic and histology-based fish health assessment, a comprehensive investigation into the fish health status of Mugil cephalus and Myxus capensis from Binfield Park Dam revealed that human effects and parasites are not the only threats to freshwater fish. Nephrocalcinosis is a non-infectious kidney disease which is characterised by abnormal calcium deposition in the kidneys of humans and some fish species. According to the macroscopic and histology-based fish health assessment, the M. cephalus and M. capensis are not in a healthy condition. The macroscopic and histology-based fish assessment indices are not stressor-specific, and therefore the cause of the poor health state of these two fish species could not be determined. A possible suggestion for the poor health of these two species is the age of the species. Because the two mullet species were stocked into Binfield Park Dam, Ellender et al. (2012) could successfully age them accurately to ten years of age, which is the upper limit of the life span for these species. Macroscopic and histology-based fish heath assessments were conducted on Labeo umbratus from Sandile Dam in order to determine the health of this species. Macroscopic and histology-based fish health assessment indicated that its L. umbratus are in a healthy state. The March 2012 survey specimens had a significantly higher macroscopic FHAI score than those from the July 2011 survey. The increased FHAI score was because of parasite infections, as well as discoloured livers and increased total blood plasma protein levels, which are indicators of nutritional state. However, the presence of the anchor worm parasite Lernaea barnimiana in low numbers had no significant impact on the health of L. umbratus. The effect of the alien anchor worm parasite Lernaea cyprinacea was shown on the transloacted small mouth yellowfish Labeobarbus aeneus. It was also shown that L. aeneus are, according the macroscopic FHAI and the histology-based fish health assessment index, in a healthy state. However, the high scores observed in the macroscopic fish health assessment index were primarily as a result of the presence of the alien parasite L. cyprinacea and its associated affects on the fish host. Because of the significant impact of this alien parasite species on the translocated host species, it can be assumed that this alien parasite species will have a negative effect on the health of indigenous fish species in the Great Kei River. It is clear from the results presented in this study that the necropsy based and histology based fish health assessment can be successfully implemented as tools to assess the effects of heavy metal pollution and alien fish parasites in freshwater fish from selected impoundments in the Amatola region, Eastern Cape Province, South Africa, thus the original hypothesis of this thesis is accepted. Based on work done in this research the gaps in research have been identified. Due to the high levels of mercury indentified in the muscle tissue of M. salmoides from Binfield Park Dam. A human health assessment and edibility should be conducted in order to determine if the fish from Binfield Park Dam is safe for human consumption. In order to conserve South Africa’s Freshwater fish biodiversity, country wide surveys of indigenous fish species must be undertaken so that the health and the parasite diversity can be evaluated. / PhD (Zoology), North-West University, Potchefstroom Campus, 2015

The conservation of biodiversity and endemism in South Africa’s freshwater aquatic ecosystems is a high priority, particularly in the Cape Floristic Region. However, the perception that South Africa lacks suitable fish species for recreational angling, aquaculture and biological control, led to the widespread introduction and use of alien fish species. As a result, formal stocking programs have seen the introduction of five of the world’s top 100 invasive species into South Africa (Dudgeon et al. 2006). According to Dudgeon et al. (2006) freshwater ecosystems are the most endangered ecosystem in the world. The threats to freshwater biodiversity, according to Dudgeon et al. (2006), can be grouped into five categories that interact with one another: overexploitation, water pollution, flow modifications, destruction of habitat and invasion by exotic species. This PhD study took place in the Amatola region of the Eastern Cape Province, South Africa. The Amatola region is a rural area with no large-scale mining or industrial developments, only localised settlements. These developments are mainly situated around impoundments, because of the resources such as water and food that they provide. Thus the dams within the Amatola region should theoretically have no major industrial stressors on them. The three impoundments studied were Binfield Park, Sandile and Wriggleswade Dams. Binfield Park Dam is a 260ha impoundment. It impounds the Tyume River and is used by both subsistence anglers from the local communities and occasionally by recreational bass anglers. Sandile Dam is a 146ha impoundment and is the smallest of the three dams in this study. It impounds the Wolf and Keiskamma Rivers. Wriggleswade Dam is a 1000ha impoundment used extensively by recreational bass and carp anglers, and impounds the Kubusi River. There is a paucity of information regarding the health of the indigenous and alien fish species from the study region, as well as on the parasite diversity of these various fish species. In order to fill the gaps in the information the following hypothesis was proposed. That the necropsy based and histology based fish health assessment can be successfully implemented as tools to assess the effects of heavy metal pollution and alien fish parasites in freshwater fish from selected impoundments in the Amatola region, Eastern Cape Province, South Africa. In order to achieve this hypothesis the main aim of this study will be to use the necropsy- and histology- based fish health assessment to determine the health status of the fish species in these impoundments as well as to understand the potential threat of water pollution and fish parasites. Fish were sampled with the aid of gill nets, fyke nets and by angling from each of the three impoundments over three surveys in July 2011, and March and August 2012. Following capture fish were transported to a field laboratory in aerated containers. At the field laboratory the fish were examined and dissected using the methods recommended by Adams et al. (1993) for a necropsy-based fish health assessment. Gills, livers, kidneys and gonads samples were also collected for histological analysis. Macroscopic and histology-based fish health assessment index was used, as well the analysis of muscle tissue of Micropterus salmoides and surface water and sediment from Binfield Park, Sandile and Wriggleswade Dams. It was shown that, according to the macroscopic fish health assessment index, M. salmoides in Wriggleswade Dam had a higher FHAI score compared to those in Binfield Park and Sandile Dam, there were no significant differences between the FHAI scores. However, the cause of the higher FHAI in the Wriggleswade Dam was because of the external skin damage caused by the presence of the alien parasite Lernaea cyprinacea. The histology-based fish health assessment index, however, showed that M. salmoides from Binfield Park had significantly higher histology Fish Index (IFISH) scores compared to those in Sandile and Wriggleswade Dams. The main contributors to the high IFISH score of Binfield Park were the significantly high Liver Index (IL) and Kidney Index (IK). The increased severity of the alterations observed in the liver and kidney tissue of the Binfield Park M. salmoides samples may have been as a result of the high concentration of mercury found in the muscle tissue of M. salmoides. The water quality and metals detected in the water of Binfield Park, Sandile and Wriggleswade Dams were all below the target water quality guideline values, as well as below those of previous research into the nutrients and presence of metals in these impoundments. The sediment metal analysis showed that the levels of Co, Mn and Ni were above the target guideline levels for Binfield Park, Sandile and Wriggleswade Dams, Cu was above target guidelines levels for Sandile Dam, and uranium was above the target guideline concentrations for Wriggleswade Dam. Binfield Park Dam had significantly high levels of mercury in the muscle tissue of M. salmoides, while Sandile Dam had significantly high levels of zinc in the muscle tissue of M. salmoides. It was shown that M. salmoides from each of the three impoundments are in a healthy state according to the parameters assessed. However, the presence of heavy metals, particularly mercury, uranium and zinc, do indicate the presence of human activities. The indigenous parasites of Anguilla mossambica have been well documented including the gastrointestinal nematode Paraquimperia africana, and the stomach nematode Heliconema africanum. Indigenous parasites such as the swimbladder nematode Anguillicola papernai had no effect on the condition factor of infected and uninfected eels. However, the damage caused by the alien parasites were evident, including the first documented effects of the alien gill monogenean Pseudodactylogyrus anguillae on indigenous wild populations of the longin fin eel A. mossambica from the Eastern Cape, South Africa. Histological observations indicated that an alien gill monogenean caused hyperplasia, increase in mucous cells, rupture of pillar cells as well as telangiectasia. This alien parasite has invaded the Keiskamma and Kei River systems in the Eastern Cape, South Africa. According to the macroscopic fish health assessment index, A. mossambica from Binfield Park, Sandile and Wriggleswade Dams are in a healthy state. However, the histology-based health assessment highlighted that the effects on P. anguillae have a severe negative impact on the health of A. mossambica. Using the macroscopic and histology-based fish health assessment, a comprehensive investigation into the fish health status of Mugil cephalus and Myxus capensis from Binfield Park Dam revealed that human effects and parasites are not the only threats to freshwater fish. Nephrocalcinosis is a non-infectious kidney disease which is characterised by abnormal calcium deposition in the kidneys of humans and some fish species. According to the macroscopic and histology-based fish health assessment, the M. cephalus and M. capensis are not in a healthy condition. The macroscopic and histology-based fish assessment indices are not stressor-specific, and therefore the cause of the poor health state of these two fish species could not be determined. A possible suggestion for the poor health of these two species is the age of the species. Because the two mullet species were stocked into Binfield Park Dam, Ellender et al. (2012) could successfully age them accurately to ten years of age, which is the upper limit of the life span for these species. Macroscopic and histology-based fish heath assessments were conducted on Labeo umbratus from Sandile Dam in order to determine the health of this species. Macroscopic and histology-based fish health assessment indicated that its L. umbratus are in a healthy state. The March 2012 survey specimens had a significantly higher macroscopic FHAI score than those from the July 2011 survey. The increased FHAI score was because of parasite infections, as well as discoloured livers and increased total blood plasma protein levels, which are indicators of nutritional state. However, the presence of the anchor worm parasite Lernaea barnimiana in low numbers had no significant impact on the health of L. umbratus. The effect of the alien anchor worm parasite Lernaea cyprinacea was shown on the transloacted small mouth yellowfish Labeobarbus aeneus. It was also shown that L. aeneus are, according the macroscopic FHAI and the histology-based fish health assessment index, in a healthy state. However, the high scores observed in the macroscopic fish health assessment index were primarily as a result of the presence of the alien parasite L. cyprinacea and its associated affects on the fish host. Because of the significant impact of this alien parasite species on the translocated host species, it can be assumed that this alien parasite species will have a negative effect on the health of indigenous fish species in the Great Kei River. It is clear from the results presented in this study that the necropsy based and histology based fish health assessment can be successfully implemented as tools to assess the effects of heavy metal pollution and alien fish parasites in freshwater fish from selected impoundments in the Amatola region, Eastern Cape Province, South Africa, thus the original hypothesis of this thesis is accepted. Based on work done in this research the gaps in research have been identified. Due to the high levels of mercury indentified in the muscle tissue of M. salmoides from Binfield Park Dam. A human health assessment and edibility should be conducted in order to determine if the fish from Binfield Park Dam is safe for human consumption. In order to conserve South Africa’s Freshwater fish biodiversity, country wide surveys of indigenous fish species must be undertaken so that the health and the parasite diversity can be evaluated. / PhD (Zoology), North-West University, Potchefstroom Campus, 2015