M.Sc. (Zoology) / A number of chemical substances in mining, industrial, agricultural and domestic effluents are likely to contaminate watercourses. These toxicants have a definite effect on all aquatic life, even at sublethal concentrations. Due to the extensive copper mining activities in the Phalaborwa region, there is concern that the sublethal effect of copper might affect the physiology of fish in the Olifants River, Kruger National Park, Transvaal. Copper can be found as an essential micronutrient in nearly all waters, although anthropogenic sources such as metals from mining and planting industries might produce environmental concentrations causing toxic effects to fish. Several effects of exposure to sublethal concentrations of copper have already been reported in fish, In the present study, the blood coagulation, general haematology, osmoregulation and differential white blood cell counts of the Mozambique tilapia, Oreochromis mossambicus were investigated after the exposures to sublethal concentrations of copper, for a short-term (96 hours) exposure as well as a long-term (four weeks) exposure in experimental flow through systems, at both 29 ± I°C and 19 ± I°C. After the respective sublethal exposures, changes in the values of blood coagulation, general haematology, osmoregulation and differential white blood cell counts were obtained, to determine the effects of the chosen copper concentrations on the blood physiology of O. mossambicus, at 29 + 10 and 19 ± I°C, respectively. After a photographic observation of blood, during all the phases of coagulation was made, it was evident that the exposure to copper, at both temperatures, resulted in delays of the blood coagulation times as well as decreases in the elasticity of the clots that formed. Copper was found to induce haemophilia at 29 ± I°C and 19 ± l°C. whilst at 19 ± I°C it also induced thrombocytopenia. Thus exposure to copper lead to coagulation defects which caused haemorrhage which can eventually cause the death of these fish...

The generation of large quantities of mine wastewater in South African coal mines and the needs for a cost effective, as well as an environmentally sustainable manner of mine water disposal, have fostered interests in the possibility of utilizing mine water for irrigation. Such a possibility will not only provide a cost-effective method of minimizing excess mine drainage, as treatment using physical, chemical and biological methods can be prohibitively expensive, but will also stabilize the dry-land crop production by enhancing dry season farming. Considering the arid to semi-arid climate of South Africa, the utilization of mine water for irrigation will also boost the beneficial exploitation of the available water resources and relieve the increasing pressure on, and the competition for, dwindling amounts of good quality water by the various sectors of the economy. The disposal of excess gypsiferous mine water through irrigation has been researched in a few collieries in the Witbank area. In this study, the assessment of the impacts of using gypsiferous mine water for irrigation were carried out in parts of the Upper Olifants basin upstream of Witbank Dam, using the ACRU2000 model and its salinity module known as ACRUSalinity. The study area was chosen on the bases of locations of previous field trials and the availability of mine water for large-scale irrigation. The primary objectives of the study were the development of relevant modules in ACRU2000 and ACRUSalinity to enable appropriate modelling and assessment of the impact of large-scale irrigation with mine water and the application of the modified models to the chosen study area. The methodology of the study included the modifications of ACRU2000 and ACRUSalinity and their application at three scales of study, viz. centre pivot, catchment and mine scales. The soils, hydrologic and salt distribution response units obtained from the centre pivot scale study were employed as inputs into the catchment scale study. The soils, hydrologic and salt distribution response units obtained from the catchment assessment were in turn applied in similar land segments identified in the mine used for the mine scale study. The modifications carried out included the incorporation of underground reservoirs as representations of underground mine-out areas, multiple water and associated salt load transfers into and out of a surface reservoir, seepages from groundwater into opencast pits, precipitation of salts in irrigated and non-irrigated areas and the incorporation of a soil surface layer into ACRUSalinity to account for the dissolution of salts during rainfall events. Two sites were chosen for the centre pivot scale study. The two sites (Syferfontein pivot of 21 ha, located in Syferfontein Colliery on virgin soils; Tweefontein pivot of 20 ha, located in Kleinkopje Colliery on rehabilitated soils) were equipped with centre pivots (which irrigated agricultural crops with mine water), as well as with rainfall, irrigation water and soil water monitoring equipment. The pivots were contoured and waterways constructed so that the runoff could leave the pivots over a weir (at Tweefontein pivot) or flume (at Syferfontein pivot) where the automatic monitoring of the quantity and quality of runoff were carried out. The runoff quantities and qualities from the pivots were used for verification of the modified ACRU2000 and ACRUSalinity. The catchment scale study was on the Tweefontein Pan catchment, which was a virgin area mainly within the Kleinkopje Colliery, draining into the Tweefontein Pan. The data on the water storage and qualities in Tweefontein Pan, as well as the soil water salinities in the irrigated area located within the catchment were used for verification of results. In the catchment scale study, different scenarios, including widespread irrigation on virgin and rehabilitated soils, were simulated and evaluated. For the mine scale study, the Kleinkopje Colliery was used. The colliery was delineated into 29 land segment areas and categorized into seven land use types, on the basis of the vegetation and land uses identified in different parts of colliery. The centre pivot and catchment scale studies indicated that the impacts of irrigation with low quality mine water on the water resources are dependent on the soil types, climate, the characteristics and the amount of the irrigation mine water applied, whether irrigation was on virgin on rehabilitated soils and the status of the mine in terms of whether a regional water table has been re-established in an opencast mining system or not. The studies further indicated that the irrigation of agricultural crops with low quality mine water may lead to increases in soil water salinity and drainage to groundwater, but that the mine water use for irrigation iii purposes can be successfully carried out as most of the water input onto the irrigated area will be lost through total evaporation and a significant proportion of the salt input, both from rainfall and irrigation water, will either be precipitated in the soil horizons or dissolved in the soil water of the soil horizons. By irrigating with a saline mine water therefore, the salts associated with the low quality mine water can be removed from the water system, thereby reducing the possibility of off-site salt export and environmental pollution. On-site salt precipitation, however, may lead to accumulation of salts in the soil horizons and consequent restriction of crop yields. Therefore, efficient cropping practices, such as leaching and selection of tolerant crops to the expected soil salinity, may be required in order to avoid the impact of long-term salinity build up and loss of crop yields. The simulated mean annual runoff and salt load contribution to Witbank Dam from the Kleinkopje Colliery were 2.0 x 103 MI and 392 tons respectively. The mean annual runoff and salt load represented 2.7% and 1.4% of the average water and salt load storage in Witbank Dam respectively. About 45% of the total water inflow and 65% of the total salt load contribution from the study area into Witbank Dam resulted from groundwater storage. From the scenario simulations, the least salt export would occur when widespread irrigation is carried out in rehabilitated areas prior to the re-establishment of the water table due to a lower runoff and runoff salt load. It may therefore be a better water management strategy in active collieries if irrigation with mine water is carried out on rehabilitated soils. In conclusion, this research work has shown that successful irrigation of some (salt tolerance) crops with low quality mine water can be done, although increases in the soil water salinity of the irrigated area, runoff from the irrigated area and drainage to the groundwater store can occur. Through the modifications carried out in the ACRU2000 model and the ACRUSalinity module in this research work, a tool has been developed, not only for application in the integrated assessment of impact of irrigation with mine water on water resources, but also for the integrated assessment and management of water resources in coal-mining environments in South Africa. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.

Water is a crucial natural resource, indispensable to food production, life, the environment, power generation, industry, sanitation and hygiene. The presence of Organochlorine Pesticides (OCPs) in the environment is not wanted due to their negative effects on human beings and animals. As a result, there is a need to continuously monitor their presence in the environment. In this study, surface water samples were collected once a month during the dry season and during wet season from the selected five points along the Olifants River and stored at a temperature of < 50C before analysis. The OCPs were extracted with dichloromethane (DCM) using the Liquid-Liquid Extraction (LLE) method. After undertaking the sample through the clean- up process, the crude extracts obtained were put into the column chromatography and eluted with hexane, about 1.5 μL of the purified extracts were analysed by the Gas Chromatographic- Mass Spectrophotometer (GC/MS). The percentage recoveries, varied from 32- 116 % for p, p’-DDT and 4,4'-DDD respectively in triply spiked water samples. The standard deviation for most of the compounds is less than±0.04, with the exclusion of Heptachlor (±0.14). The seasonal variability of OCPs in water samples along Olifants River results show that in dry season, the Olifants River is mostly polluted at the Oxford site with (BHC-beta, Aldrin, Heptachlor-epoxide, Endosulfan-alpha and Endrin), at the Ga- Selati site with (Heptachlor-epoxide and Endrin) and at the Wolvekrans site with (Endosulfan-alpha), with Aldrin up to 834.20 ng/ L indicating the highest hazard toward the aquatic environment while in summer the Olifants River is mostly polluted at the Ga- selati site with BHC-beta and at the Waterval site with (Heptachlor and BHC-gamma) with BHC- gamma up to 560 ng/ L indicating the highest hazard toward the aquatic environment. The levels reached from the Olifants River catchment were meaningfully above the drinking water quality guidelines for organic chemical recommended by WHO, 2006 i.e. (BHC-gamma,DDT-44, Aldrin, dieldrin and Endrin are (2.0,1.0,0.03,0.03 and 0.6) respectively for the protection of the domestic use, aquatic ecology and agricultural use (irrigation and livestock watering) for compounds with local guideline values; while, the international water quality guidelines to protect the aquatic ecosystems are 0.00083 ng/mℓ (4,4’DDD), 0.00059 ng/mL (4,4’DDE), (4,4’DDT), 0.00021 ng/mL (heptachlor), 0.0092 ng/mℓ (α-HCH), 0.0186 ng/mL (γ- HCH), and chronic values are 0.056 ng/mL (ENDO I and II) and 0.0023 ng/mL endrin) (USEPA, 2002). Levels detected were significantly higher than some research studies conducted up to now in South African aquatic environments. These results confirm the contamination of the Olifants River catchment by the OCPs. / Water and Sanitation / M. Sc. (Environmental Science)

M.Sc. / Pollution of the earth is presently one of the most important environmental issues of the world and various attempts, including creation of public awareness have been initiated over the past few years to address this situation. There is especially a global concern about the progressive pollution of valuable freshwater systems which most organisms, including humans, are reliant upon. This includes South Africa's aquatic ecosystems, many of which have been degraded over the past few decades. The Olifants River in Mpumalanga is one of the most threatened river systems in South Africa. It is impacted by various anthropogenic activities, particularly mining in the upper catchment, and urbanisation, industrial and agricultural activities in the upper and lower catchments. Previous investigations have shown that these activities are responsible for the degradation of this river system. The broad objective of this study was to obtain data on water and sediment quality, as well as bioaccumulation of metals in fish via monitoring of the Olifants River. It formed part of a major study concerned with the experimental investigation of lethal and sub-lethal effects of metals on fish physiology, supported by a field investigation into the water quality and metal contamination of the biotic and abiotic components of the Olifants River system. Physico-chemical properties of the water as well as the concentrations of some metals (Al, Fe, Cu, Zn, Ni, Mn, Pb & Cr) in the water and sediment were seasonally investigated. The extent of bioaccumulation of these metals in selected organs/tissues (muscle, gills, liver & skin) of Oreochromis mossambicus and Clarias gariepinus from Loskop Dam (upper catchment) and Mamba Weir, Kruger National Park (lower catchment) was also investigated. High levels of various macro-constituents and metals were detected at many sites in the study area and in many cases they exceeded the guideline limits set for aquatic ecosystems. Localities 2, 3, 6, 8, 10, 12 and 17 were observed to be severely impacted (see Chapter 9- Table 9.1) by elevated concentrations of pollutants influencing variables such as TDS and sulphates. This confirms that these sites are being impacted by mining. This was further confirmed by low pH-values at localities 3, 5, 8, 9, 10 and 12 which indicate on acid mine drainage originating from the many coal mines in the upper catchment of the Olifants River. Nutrient enrichment (elevated levels of phosphates, nitrates and nitrites) occurred at many sites in the catchment but in particular at localities 3, 4, 6, 10, 11, 14, 15 and 17. Point source pollution from sewage treatment works and non-point sources from agricultural runoff and informal settlements are the main contributors to these elevated levels of nutrients. The Selati River, impacted in particular by elevated phosphate levels, is the main contributor to the high nutrient levels detected at locality 17. It is evident from the evaluation of the metal concentrations in the water and sediment (Table 9.2) that most of the sites in the Olifants River catchment are being impacted by metal pollution. Oreochromis mossamhicus and C gariepinus sampled at selected sites in the Olifants River accumulated selected metals in the following order; Fe>Zn>Al>Cr>Ni>Pb>Mn>Cu. Levels were generally high in the liver and gills, while much lower levels were detected in the skin and muscle tissues. Gill tissue is in direct contact with the aqueous environment and therefore gives a good indication of the extent of exposure. Skin tissues usually contained relatively low levels of metals (except for Zn). Although muscle tissues usually accumulated low metal concentrations it is an important tissue to monitor in bioaccumulation studies as it can lead to metal poisoning if contaminated muscle tissues are consumed by humans. Adult specimens were mostly sampled and within this range there was a slight decrease in Cu, Al, Fe, Ni, Mn and Cr concentrations with increasing age. Temporal variation in metal accumulation by fish possibly occurred due to variations in metal concentrations in the water and sediment at a locality. These differences were caused by seasonal variation in climatical conditions (eg. rainfall, temperature), as well as temporal fluctuations of pollutant inputs into the river system. Aquatic organisms at both localities 15 (Loskop Dam) in the upper catchment and 17 (Mamba weir, Kruger National Park) in the lower catchment are at present being chronically exposed to elevated levels of the investigated metals, compared to both the control site and results in literature. The impact of the highly polluted Selati River in the lower catchment was evident in the difference between metal concentrations detected in fish at locality 17, and at locality 19 (Phalaborwa Barrage) upstream of the Olifants-Selati confluence. The present study clearly indicates that the Olifants River is subjected to various sources of pollution which could be detrimental to the health of this aquatic ecosystem. It is recommended (see Chapter 9) that a multi-disciplinary approach including a biomonitoring programme, should be followed to ensure a sustainable freshwater ecosystem.

D.Phil. / The entire Olifants . River Catchment is subjected to increasing afforestation, agricultural, domestic, mining, industrial, irrigation and urbanisation activities. These activities have a profound effect on the water quality of the river and its tributaries. This is cause for concern for the water users in the upper catchment, and because the Kruger National Park, one of its downstream water users, is extremely dependent on water of a satisfactory quality to sustain its various ecosystems. It is therefore vitally important to determine to what extent activities in the Upper Catchment of the Olifants River (Mpumalanga), especially in the Witbank and Middelburg areas, influence the water quality of the river. Point sources of pollution in the upper reaches include mining and industrial activities as well as water care works located at various points along the river. These were addressed in a Water Research Commission Project (No. 608/1/97) titled "Lethal and sublethal effects of metals on the physiology of fish: An experimental approach with monitoring support". Although sixteen localities were chosen for the initial project, this study only focussed around the metal ecotoxicology at two localities (Steenkool Spruit and Witbank Dam) in the upper catchment and the effect of metals (copper and zinc) on fish blood physiology. In aquatic ecosystems water quality is an important variable and full assessment of water quality, of Steenkool Spruit (locality 3) and Witbank Dam (locality 7), included evaluation of the chemical, physical and biological characteristics at each of the localities. Water and sediment samples were collected seasonally during the study period, February 1994 to May 1995, and the chemical and physical water quality variables were measured. During the study period three metal bioaccumulation indicator species, Labeo umbratus, Clarias gariepinus and Labeo capensis, were captured from which tissue (gills, liver, muscle and skin) samples were collected. These samples as well as water and sediment samples, were analysed for aluminium, chromium, copper, iron, manganese, nickel, lead and zinc concentrations, using atomic absorption spectrophotometry. In the past routine monitoring of chemical and physical water quality characteristics left scientists and managers with a sizeable pool of data which is often difficult but not impossible to interpret. To standardise and summarise this collection of data an aquatic toxicity index (ATI), WATER2 was developed by Wepener et at (1992). The present study attempted to expand and refine WATER2, which has resulted in the establishment of a new ATI, RAUWater.

Since 2008, large numbers of Nile crocodile (Crocodylus niloticus) carcasses were found in the Kruger National Park (KNP), South Africa. Most of the crocodile carcasses were found in the Olifants Gorge, which is situated below the Letaba and Olifants river confluence, before the Mozambique border and Massingir Dam. The Massingir Dam is an important resource and it plays a significant role in the welfare of the local Mozambican population. Autopsies performed on the crocodiles indicated that the adipose tissue colour changed from normal white to yellow and this is usually a sign of pansteatitis. Pansteatitis is caused by lipid peroxidation in an organism and it is characterised by the lack of vitamin E. This disease is recognisable by the hardening of the fatty tissue and yellow discolouration, and is mostly associated with aquatic organisms from polluted ecosystems. There are speculations that the crocodile fatalities may be associated with the Massingir Dam that backed up into the Olifants Gorge after flooding. After the dam was reconstructed, it flooded the Olifants Gorge, causing it to act like a localised sediment trap as the water flow slowed down and as a result, caused pollutants to build-up. Sediment samples were collected from selected rivers and ponds within the KNP. These samples were analysed for selected elements, persistent organic pollutants (POPs), and polycyclic aromatic hydrocarbons (PAHs). The sediment samples were analysed in Norway for POPs and PAHs with the use of a high-resolution gas chromatography/mass spectrometry (GC/MS) and the heavy metals were analysed in South Africa with the use of inductively-coupled plasma mass spectrometry (ICP/MS). In order to identify which elements may have affected the health of the crocodiles, a series of sediment quality indices were used. These indices made it possible to determine which elements may have been involved. The order of probability of heavy metals causing harm was Se>As>Ni>Cr>Cu>I>V>Mn>Co>Fe>Cd>Hg>Zn>Pb>Ba>U. The data was compared to selected international guidelines. All the information was used to determine which of the sampled sites had the highest contamination. The sites sampled with the highest concentrations were in the Crocodile, Nkomati, Olifants, and Letaba Rivers. Concentrations of the elements, POPs, and PAHs were also quantifiable in the Olifants Gorge. The following elements (Fe, Co, Cu, Cr, Pb, V, As, and Ni) were quantified at elevated levels and may therefore have caused negative effects on the crocodiles in the Olifants Gorge. These elevated concentrations, in combination with the dramatic change in the physical environment due to the dam, could have added additional stress that may have contributed to the observed crocodile mortalities in the Olifants Gorge. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Since 2008, large numbers of Nile crocodile (Crocodylus niloticus) carcasses were found in the Kruger National Park (KNP), South Africa. Most of the crocodile carcasses were found in the Olifants Gorge, which is situated below the Letaba and Olifants river confluence, before the Mozambique border and Massingir Dam. The Massingir Dam is an important resource and it plays a significant role in the welfare of the local Mozambican population. Autopsies performed on the crocodiles indicated that the adipose tissue colour changed from normal white to yellow and this is usually a sign of pansteatitis. Pansteatitis is caused by lipid peroxidation in an organism and it is characterised by the lack of vitamin E. This disease is recognisable by the hardening of the fatty tissue and yellow discolouration, and is mostly associated with aquatic organisms from polluted ecosystems. There are speculations that the crocodile fatalities may be associated with the Massingir Dam that backed up into the Olifants Gorge after flooding. After the dam was reconstructed, it flooded the Olifants Gorge, causing it to act like a localised sediment trap as the water flow slowed down and as a result, caused pollutants to build-up. Sediment samples were collected from selected rivers and ponds within the KNP. These samples were analysed for selected elements, persistent organic pollutants (POPs), and polycyclic aromatic hydrocarbons (PAHs). The sediment samples were analysed in Norway for POPs and PAHs with the use of a high-resolution gas chromatography/mass spectrometry (GC/MS) and the heavy metals were analysed in South Africa with the use of inductively-coupled plasma mass spectrometry (ICP/MS). In order to identify which elements may have affected the health of the crocodiles, a series of sediment quality indices were used. These indices made it possible to determine which elements may have been involved. The order of probability of heavy metals causing harm was Se>As>Ni>Cr>Cu>I>V>Mn>Co>Fe>Cd>Hg>Zn>Pb>Ba>U. The data was compared to selected international guidelines. All the information was used to determine which of the sampled sites had the highest contamination. The sites sampled with the highest concentrations were in the Crocodile, Nkomati, Olifants, and Letaba Rivers. Concentrations of the elements, POPs, and PAHs were also quantifiable in the Olifants Gorge. The following elements (Fe, Co, Cu, Cr, Pb, V, As, and Ni) were quantified at elevated levels and may therefore have caused negative effects on the crocodiles in the Olifants Gorge. These elevated concentrations, in combination with the dramatic change in the physical environment due to the dam, could have added additional stress that may have contributed to the observed crocodile mortalities in the Olifants Gorge. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

M.Sc. / Metal ions have become an increasing source of pollution in the natural waters of South Africa. This phenomenon is related to the rapid industrial, mining and agricultural activity growth along rivers, especially in the Upper catchment of the Olifants River, Mpumalanga. In general, metal ions have - negative effects on the physiology of fish and other aquatic biota in metal polluted waters. In this study. the sublethal effects of, manganese were determined by exposing the freshwater fish, Oreochromis mossambicus, to this metal in an experimental flow-through system. The exposure times were divided in acute (96 hours) and chronic (26 days) exposures, both at 23 ± 1°C. The results obtained showed changes in the general, haematological, metabolic, osmoregulatory variables, as well as the differential white blood cell counts. These changes indicated several effects in the physiology of a mossainblais after sublethal manganese exposure. Significant differences (P < 0.05) were found between the haematological variables of the control and exposed fish. During chronic exposure, an oxygen deficiency developed due to , the epithelial lifting of gill lamellae. As a result of hypoxia the red blood cell numbers, haemoglobin-, haematocrit and lactate concentrations increased to uplift the depleted oxygen situation. The significant differences (P < 0.05) found in the sodium, potassium,. calcium and chloride concentrations are mainly as a result of gill damage. Manganese showed a disruptive influence on the structural organization of the gill. The differential white blood cell counts performed , showed significant fluctuations. This leucocytosis and leucopenia are normal reactions of the fishes’ body against infections of foreign substances, such as metal ions. It was concluded that the general haematology, metabolism, osmoregulation and differential white blood cell counts can, be used as indicators in detecting the effects of sublethal manganese exposure on fish.

M.Sc. (Zoology) / Please refer to full text to view abstract

M.Sc. (Environmental Management) / The Kruger National Park (KNP) is a world renowned wildlife reserve and a source of South African ecotourism benefiting the economy. The Olifants River is the largest river running through the KNP and it is known to be one of South Africa’s most polluted rivers. In the winters of 2008 and 2009 Nile crocodile carcasses were found in the Olifants River gorge in the KNP. In a very short period nearly the entire population of Nile crocodile was lost, the cause of which was later identified as pansteatitis. Pansteatitis is caused by lipid peroxidation known to be nutritionally mediated, as it is associated with diets high in polyunsaturated fats, often of fish origin and insufficient antioxidants specifically vitamin E, which is solely synthesized by plants and must therefore be obtained through diet. The hypothesis of this study is therefore that the fish inhabiting the Olifants River are antioxidant deficient, more specifically, vitamin E deficient, causing the wildlife e.g. crocodiles and predatory fish species, that feed on the fish, to become vitamin E deficient as well, and subsequently develop pansteatitis. If the hypothesis is true, the top fish predator in this aquatic system, the tigerfish Hydrocynus vittatus, should therefore also exhibit signs of dietary vitamin E deficiency. The aim of this study was to determine if tigerfish from the Olifants River in the KNP are exhibiting any signs of antioxidant deficiency, specifically vitamin E deficiency. The objectives to accomplish this aim were firstly to analyse the total plasma antioxidant activity using an ELISA assay, and secondly, to perform a histology-based fish health assessment on the target organs of the tigerfish to identify any histological alterations, specifically those known to be associated with vitamin E deficiency. The results were compared to the results from fish of the same species from two reference sites where there have been no signs of vitamin E deficiency, such as pansteatitis-related wildlife deaths. The results of the plasma analysis as well as the histological assessment showed no conclusive signs of vitamin E deficiency in tigerfish from the Olifants River. It is therefore unlikely that the pansteatitis in the affected wildlife is caused by dietary vitamin E deficiency in the aquatic system, but rather by vitamin E depleting mechanisms such as the presence of high amounts of toxicants with pro-oxidant properties, which may be causing an imbalance of pro-oxidants and antioxidants within the body. Hence the rate of vitamin E regeneration cannot keep up with the rate of oxidation. This is occurring only in certain species due to their specific behaviour or eating habits.