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

M.Sc. (Zoology) / The catchment area of the Olifants River is being subjected to increased agricultural and mining activities, industrial development and urbanisation. As a result of this, the water quality 'of the Olifants River and some of its tributaries (e.g. Selati River) has been deteriorating since 1983. This causes reason for concern as one of the downstream water users in the Olifants River catchment is the Kruger National Park, which requires of good quality to sustain its terrestrial and aquatic ecosystems It was therefore necessary to determine what extent activity midstream-or--tlie I Olifants River, especial y in the Phalaborwa area, influenced the water quality of the Olifants River. In this study, an evaluation was done of the water quality (physically and chemically of the Lower Olifants River inside of the Kruger National Park, as well as the Lower Selati River, a tributary of the Olifants River which flows through the Phalaborwa area. Special attention was paid to the metal concentrations in the water, sediment and fish. Water and sediment were sampled every alternate month from April 1990 to February 1992 at six sampling sites along the Lower Olifants River and one in the Selati River. Standard methods were used to determine the physical and chemical characteristics (e.g. pH, TDS, etc.) of the water. The fish species Barbus marequensis was also sampled every alternative month, but only at three sampling sites along the Olifants River and one in the Selati River. Organs and tissues that were dissected, included the gills, fat, liver, gonads, gut, muscle, skin, blood, vertebrae, kidney and bile, as well as the gut contents. In February 1992, additional sampling was performed at Pioneer Dam, a natural reference point used in this study. Atomic absorption spectrophotometry was used in the laboratory to determine the Cr, Cu, Fe, Mn, Ni, Pb, Sr and Zn concentrations in the water, sediment and fish samples. (The water quality of the Selati River was found to be stressful to aquatic life, especially .) with regard to the sodium, fluoride, chloride, sulphate, potassium, total ~dissolved salts 1 and metal concentrations (except strontium). The Selati River also influenced the ': water quality of the Olifants River after the Selati-Olifants confluence. Most of the :. time the water quality of the Lower Olifants River in the KNP did, however, comply With the recommended guideline limits, except for the metal concentrations. The high metal concentrations detected in the water (in some cases sub-lethal levels) indicated some degree of metal pollution, but, due to the hardness of the Olifants River water, conditions were not necessarily toxic to the aquatic life. The accumulated metals in the organs and tissues of marequensis gave a good indication of the metal levels to which the fish were exposed. More metals were taken up by the fish from April 1990 to February 1991 than from April 1991 to February 1992. The various metals were . distributed differently to the tissues of B. marequensis, with the highest zinc concentrations being accumulated by the skin and ovaries; the highest copper and iron by the liver, kidney and gut; the highest chromium and nickel by the blood; and the highest manganese, lead and strontium by the vertebrae and gills. These results afforded guidelines as to the types of tissue which should be sampled as the most suitable for the analysis of the various metals in polluted waters.

M.Sc. / The catchment area of the Olifants River has, over a number of years, been exposed to extensive mining, agriculture and urbanisation activities in the Witbank-Middelburg and Phalaborwa regions which has largely contributed to the deterioration of water quality. Of major concern is the influence anthropogenic activities have on the aquatic ecosystem of the Olifants River within the Kruger National Park and how me health of fish residing In these waters is affected. To determine fish health, a biological monitoring method, the Health Assessment Index (H41), was tested for the first time In South Africa. Four surveys were conducted at Mamba and Balule In the Olifants River, Kruger National Park during 1994 (February, May, July and November). An additional survey was conducted in February 1995 at Loskop Dam. Samples of water and sediment were taken for analysis of metals and physical and chemical water parameters. A maximum of 20 Clarias gariepinus fish were sampled at each location. Evaluation of the fish was done according to guidelines set in the HAI and parasite population composition (prevalence, abundance, mean intensity) was determined. Organ and tissue samples including gills, liver, muscle and skin were analysed for the bioaccumulation of chromium, copper, Iron, manganese, nickel, lead, strontium and zinc, using atomic absorption spectrophotomeby. Metal concentrations in the water at Mamba and Balule were within guideline limits, whereas concentrations at Loskop Dam were above guidelines. As reflected by the application of the Aquatic toxicity Index, Loskop Dam presented with the poorest water quality followed by Mamba then Balule. It was found that certain physical and chemical variables namely fluoride, potassium, sulphate and total dissolved solids concentrations at Mamba and Balule were relatively high, particularly during drier months. Metals accumulated in organs and tissues, with the highest concentrations In the gills followed by the liver, skin and muscle. The discriminant analysis, utilising metal bloaccumulation, discriminates between water quality at Mamba and Balule, revealing a 100 % classification probability for each survey. Values obtained In the application of the HAI indicated that variables with good predictor accuracy were plasma protein, all parasites, endoparasites, liver, white blood cell counts, ectoparasites, skin, fins and gills. The discriminant function for the HAI generally Indicated variables similar to those exhibiting high predictor accuracy. The discriminant function showed relatively low classification probability for each survey. In case 1, where separate endo- and ectoparasite variables were Included in the determination of me discriminant function, probability for me entire study ranged between 47.5 % and 84.2 %. In case 2, where endo- and ectoparasite variables were given a refined score rating system, probability ranged between 62.5 % and 100 %. The low classification probability Indicates either the Importance of repetitive testing for this technique or a total departure from it. Results showed that fish populations with higher HAI values are found in water of poorer quality (Mamba), while healthier fish populations i.e. with lower RAI values are found In water of better quality (Balule). Parasite data shows a similar tendency by indicating that ectoparasites are abundant in water of higher quality, while endoparasftes increase in water of poorer quality. The HAI reflects the condition of fish populations in relation to their aquatic environment, therefore, the HA/ gives an indication of water quality and should be used as a first level screening tool. If complemented by a parasite survey, distinguishing between endoand ectoparasites, the results from the HAI will be enhanced.

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. / The recognition of the occurrence, importance and effects of contaminants on food chains and ecosystems has led to the development of biomonitoring programmes that use indicator species to estimate the levels in other parts of the ecosystem. There has been an increased awareness of potential pollution of ecosystems in Gauteng, South Africa, because of increased formal and informal urbanization, industrialization and mining activities. It is essential to obtain clear indications of metal pollution cause-effect relationships at ecosystem level in order to carry out effective management of these ecosystems. The objective was to investigate the application of an ecotoxicologically-based investigation of metals in selected bird species as bioindicators of metal pollution of ecosystems in Gauteng. Three passerine species were used, the Southern Masked Weaver (Ploceus velatus), the Red Bishop (Euplectes orix) and the Red-billed Quelea (Quelea quelea). The sites were selected to represent a theoretical pollution gradient from severely contaminated wetlands in the Vaal Triangle, to moderately contaminated wetlands in the Witwatersrand and Pretoria. A nature reserve in North West Province was chosen as reference site. The sites were Rietvlei in Tshwane, Roodekrans and Olifantsvlei in Johannesburg, Holfontein in the Vaal Triangle and Barberspan in North West Province. Two sampling surveys were carried out, one in 2002 and one in 2005. Feathers were collected for metal analyses. Blood samples were collected for biomarker (oxidative stress and DNA damage) and haematological analyses. The samples were digested using the microwave destruction technique. The metal analyses were carried out using ICP-MS. Oxidative stress enzymes (reduced glutathione content and catalase activity) were analysed in plasma samples and DNA damage was evaluated in red blood cells using the average base-pair length comparisontechnique. Haematological studies were done on the blood samples. The results were compared between the sites and surveys. The levels of stress correlated well with the levels of metals in the feathers. Sites with higher metal levels had higher levels of organism stress. Thus the study showed that the feathers can indeed be used in biomonitoring. Internationally there is a trend to incorporate Ecological Quality Objectives (EcoQOs) into legislation, especially when dealing with metal contamination. Within the framework of EcoQOs developed for birds the monitoring of contaminants forms a specific category of EcoQOs. However, due to a lack of available data, Reference and Target Levels still need to be set. The development of EcoQOs is especially important at this stage in South Africa. The recently promulgated National Biodiversity Act (10 of 2004) specifically provides for the preparation of conservation plans for identified ecosystems that are important because of their goods and services that they provide. Since there is no management tool available at present to develop a suitable management plan, the setting of EcoQOs that are in line with international standards and practices will be of immeasurable value to implementing the Act. Feathers from museum specimens were used to determine the Reference Levels for the various metals in the feathers. The Target Levels were calculated from the Reference Levels. These were compared to the Current Levels and the EcoQOs were determined for the metal levels in the feathers in weaver in Gauteng, South Africa. / Prof. V. Wepener

M.Sc. / A comparative study was undertaken in the Olifants River catchment, to determine the water quality at two dam sites, namely, Bronkhorstspruit Dam (control) and Loskop Dam (polluted) in the upper Olifants River system and two river point, namely, at Mamba and Balule in the Kruger National Park, in the lower Olifants River system. Data was obtained during the physical and chemical analysis of water and sediment samples, and during bioaccumulation studies using Atomic Absorption spectrometry, testing for aluminium, chromium, copper, iron, lead, manganese, nickel, strontium and zinc. These tests were conducted on the liver, skin, muscle and gills of Clarias gariepinus. The information collected from Mamba and Balule, was compared with a previous study carried out primarily in the Kruger National Park by Marx, (1996). The study undertaken by Marx (1996) was carried out during a drought period, which allowed for the comparison with results recorded after flooding during the present study. This current research project therefore allowed for the comparison between two different water sources, namely, that of dams and rivers, and under varying environmental conditions. The Aquatic Toxicity index (ATI) developed by Wepener et al. (1992) was employed to facilitate the comparison between the water physical and chemical parameters that were measured, at each sample site, with a single variable being calculated for each sample site per survey. The water quality (ATI values) at the two control sites namely, Bronkhorstspruit Dam and Balule deteriorated substantially after the floods. High water levels due to the floods had a dilution effect on the concentration of pollutants however, toxicants were washed in from upstream and due to surface runoff. The removal of the purifying reed beds upstream of Balule and at the inflow to Bronkhorstspruit Dam, resulted in the release of latent sludge containing metals and organic pollutants into the water column. The ATI value's obtained for the two control points were similar or even higher than those obtained for the two polluted sites namely at, Loskop Dam and Mamba for the autumn and winter sampling periods. During following surveys the water quality at all four sample sites improved, returning to pre flood values by the last survey in summer, namely reflecting similar values as those recorded by Marx (1996) at Balule, Mamba and Loskop Dam for the same period. Sediment concentrations recorded a similar trend to that for water at all four sample sites throughout the study. The bioaccumulation study indicated that the gill concentrations recorded were generally the highest, followed by the liver concentrations. From this one may deduce that gills were the dominant site for metal absorption by the fish, with the excretion of metals also taking place via this route. Liver concentrations are an indication of the activation of protective _mechanisms in the fish sampled, the high concentrations recorded indicate the high physiological response the fish have had to the various levels of exposure. The low concentrations recorded in the muscle indicates the effectiveness of the liver in the detoxification of the fish, indicating that only limited storage of the various metals tested for took place at this site. Skin concentrations were quite high, these concentrations represent the products of an excretory process, which takes place via the skin. The order of concentrations recorded during the present study, were similar to the orders recorded by Marx, (1996) and other authors, however, the order of the skin and muscle concentrations were often reversed. This may be due to the increased excretion of these metals via the skin, resulting from increased exposure to these metals after the floods. Thus the increased exposure

D.Phil. / By virtue of its position on the eastern border of South Africa, the Kruger National Park (KNP) receives the flow of six main riven, of which the greater part of the specific catchment areas are situated upstream from the park itself. The Olifants River has the largest catchment area both in size and in proportion of catchment area beyond the park boundary. Increased competing demand for water by the urban, industrial, mining and agricultural sectors within the catchment boundaries of the Olifants River has focused attention on the ecological water requirements of the river. Of particular concern was the high degree of mineralisation of the Olifants River due to mining activities on the western boundary of the KNP. When one considers the demand for water by the Kruger National Park it is evident that water is required for more than one use, i.e. for potable use, game watering and ecosystem (aquatic and riparian) maintenance. In order to facilitate proper water quality management it is necessary to develop a comprehensive water quality management plan for the river. The development of such a plan requires a thorough understanding of the fate and effects of pollutants in the environment. It is furthermore important to have reliable information on the trends and status of important water quality determinants in these systems. This study was therefore aimed at investigating the metal ecotoxicology of the lower Olifants River and the Selati River, a tributary of the Olifants River which flows through the Phalaborwa area. Water quality is a driving variable in aquatic ecosystems. Full assessment of water quality of the Olifants River included evaluation of the physical, chemical and biological attributes of the system. The difference between the physical/chemical and biological attributes is that the former provide an instantaneous and often specific assessment of the water quality, whereas the latter integrate changes in the system over time. Water and sediment samples were collected bi-monthly from February 1990 to February 1992 at six stations along the lower Olifants River, one in the Selati River and one in the Letaba River. At each of these sites physical and chemical water quality characteristics were measured. Two metal bioaccumulation indicator species, Labeo rosae and Oreochromis mossambicus, were also sampled during the same period at three stations in the Olifants River and at the Selati River sampling station. Two additional surveys were undertaken to reference sites in the KNP (i.e. Pioneer Dam, and Nhlanganini Dam) during February 1992 and April 1992 respectively. Tissue samples (muscle, gill, liver, gonads, fat and blood) were collected and analysed for Cr, Cu, Fe, Mn, Ni, Pb and Zn using atomic absorption spectrophotometry. The routine monitoring of chemical and physical water quality attributes produces large quantities of complex data which are often difficult to interpret. To reduce the complexity, data may be standardised and summarised into an index. The Aquatic Toxicity Index (ATI) that was developed Metal Ecotoxicology in the Olifants River it Summary during this study presented the water quality of all the sampling stations, during each survey as a single value. This value reflected the water quality's suitability-for-use by fish at the different sampling stations. The ATI showed definite temporal and spatial trends in water quality in the Olifants River. The Selati River was the main contributing factor to the poor water quality measured in the Olifants River within the KNP. This was due to high TDS, metal and fluoride concentrations. Deterioration of the water quality was experienced during flood conditions when the turbidities and total metal concentrations increased. The water quality generally increased between summer and winter surveys (February to June). Bioindicators are species used to quantify biologically available level of metals in the aquatic ecosystem. These organisms take up metals from the ambient water and from food, and the concentrations in their tissues (or changes in concentrations) provide a time integrated measure of metal bioavailability. Temporal bioavailability of metals in the tissues of the bioindicators showed similar trends to the water quality. Highest metal concentrations were measured in fish tissue samples collected during flood conditions (December 1990) and during low-flow conditions (June to October). This was attributed to increased exposure due to metal adsorption to suspended sediments during the flood conditions and to increased ambient metal concentrations during lowflow conditions. The distribution of metals in tissues varied based on the uptake, depuration and metabolism strategies for each metal. In general the gill tissue bioaccumulated the highest metal concentrations, with only Cu and Fe occurring in higher concentrations in liver tissue. Significant positive correlation's were found between bioaccumulation in muscle tissue and the other tissues: Space-bulking and time-bulking of results were carried out to elucidate spatial bioaccumulation patterns. The highest bioaccumulation of Cu, Pb and Ni were recorded in samples from the Selati River, whereas Zn and Cr concentrations were the highest in the Olifants River. An equilibrium partitioning (EP) technique was used to integrate bioaccumulation data and metal concentrations in different phases of the water and sediment and produce specific Cu and Zn criteria for the Olifants and Selati rivers. Although there is a multitude of data available for Cu and Zn toxicity and BCF's, the quality criteria produced during this study, for the first time, made use of results obtained under local conditions. The results from bioaccumulation studies carried out in the field can not provide information regarding the potential effects or the risks the metals may pose to the exposed organisms. Therefore the susceptibility of the organisms to metal contamination can be determined using laboratory studies. Physiology is toxicology's closest relative and contributes markedly in assessing the toxicological risk of metals. Bioassays were conducted to determine the uptake kinetics, acid-base disturbances and haematological effects of a metal mixture containing Cu, Fe and Zn on Tilapia sparrmanii. The effects of the individual metals (Cu, Fe and Zn) and different Metal Ecotoxicology

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...

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.