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M. Tech. Biomedical Technology. / In most rural areas of developing countries, people use untreated water sources for consumption and sanitation purposes. The current health-related water quality testing techniques only determine the presence of indicator organisms and may not adequately predict the potential of water to result in adverse physiological effects after ingestion. The possibility of using an accurate and reliable in vitro bioassay that could directly predict the potential of water to cause an infection after ingestion was investigated in this study.

Prof. Paul Jagals Dr. Hafsatou Ndama Traoré

M. Tech. / The study assessed the risk of infection introduced by containers in which households collect water from different sources. The study area was in rural villages in the Vhembe region of the Limpopo Province. Each village had its own unique water sources consisting mainly of untreated ground waters and untreated surface waters. Randomly selected household members use their containers to collect water from these different sources. Two of the three villages received new small water supply systems within the study period, proving the villagers with water of good health-related microbial quality. The remaining village continued to use contaminated water from their untreated surface water sources. The water supply intervention in two of the three villages provided the opportunity to assess the impact these interventions would have on the risk of infection i.e. whether the risk could be reduced for the villagers no having access to good quality water. A customized quantitative microbial risk assessment was done based on the health-related quality of the water that people ingested before and after the intervention. This assessment was based on exposure of, consequence to and impact on an individual water consumer. Exposure included variables such as daily quantities of unheated drinking water consumed per individual (in litres) available at the point of use (in the dwelling) for ingestion, numbers of diarrhoeagenic E. coli (Dec) per litre in water sampled from containers derived from water samples positive for indicator E. coli and daily doses of DEc per litre ingested by an individual in the target population. Consequence was assessed by calculating the probability of infection per day from diarrhoeagenic E. coli for an individual and from there the annual infection per individual. The impact of the small water supply system was determined by assessing changes in annual individual risk of infection from before to after the interventions per village. In terms of daily water volumes consumed per individual, there were no significant differences in consumption amongst the three villages. At 1.26 ℓcd (at the upper limit of the 95% confidence interval) the consumption was slightly higher than the one litre suggested by WHO 2003. The container water collected from sources before the intervention in the two villages was significantly more contaminated by indicator E. coli (iEc) and diarrhoeagenic E. coli (DEc) than their container waters after the intervention. In terms of impact, the risk of infections was substantially reduced by the interventions. For Village 1 the risk of infection was reduced from 646 infections to 135 infections per 10,000 of the population. If these are converted to 10% as disease manifesting, then there could have been 65 cases of enteric disease attributable to water before the intervention, which was then reduced to 14 cases after the intervention. For Village 2 the population risk was reduced from 110 to 67 cases after the intervention. For Village 3 the population risk remained at 2,778 infections or 278 cases of enteric waterborne disease because of their continued use of untreated water. The study findings indicated that for Village 1, the population risk of infection would be 135 infections per 10,000, for Village 2 it would be 167 and for Village 3 - 234. If the hypothetical conversion rate of one case of enteric waterborne disease for every 10 infections of Howard et al. (2006) is used, then this would mean 14 cases of disease per 10,000 for the population for Village 1, 17 cases for Village 2 and 24 cases for Village 3. Relating this to the WHO (2003) suggestion of one case per 1,000 as an acceptable, it would mean 1.3 cases for Village 1, 1.7 for Village 2 and 2.4 for Village 3. For Village 1 the risk of contracting waterborne enteric disease was close to acceptable, but not at all acceptable for the other two villages, even for Village group 2 after the intervention. The water system in Village 2 failed often during the study which was probably the cause of the persistently higher risk of disease. It can be concluded that providing a well maintained small water supply system reduced the risk to and maintained it at an acceptable level. Table

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Water is essential to all life but many freshwater resources are polluted through human activities. Humans and wildlife are exposed to a wide range of contaminants through their water, many of which pose a risk to health. Some of the contaminants released into the environment have been reported to have the capability to disrupt the endocrine functions in humans and wildlife and they can mimic or antagonise the action of estrogenic. These endocrine disrupting chemicals (EDCs) interact with physiological systems and cause alterations in development, growth and reproduction in wildlife and humans. To achieve some measure of assessing the potential harm that the contaminants pose, we need to know the environmental concentration of the chemical concerned and to monitor their effect on the organisms. The water supply sector need to include EDCs in standard systems of routine water source monitoring which include indicator bacteria and nutrient species but before the system can be incorporated, methods to measure the occurrence of EDCs in aquatic environment need to be developed and validated and a reliable guidelines data need to be in place. The aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA) to quantify vitellogenin (VTG) in Oreochromis mossambicus (Mozambique tilapia) VTG has been used successfully as a biomarker for estrogenic contamination in different studies. For this study, VTG was isolated and purified from plasma of 17β-estradiol exposed tilapia by gel filtration chromatography. The purity of the VTG isolate was confirmed by polyacrylamide gel electrophoreses (SDS-PAGE). Polyclonal antibodies against t-VTG were raised in rabbits and the specificity of the anti-t-VTG was confirmed by western blot. Using purified t-VTG as a standard and anti-t-VTG antibody, a homologous competitive ELISA was developed and validated. The standard curves of the ELISA, which were generated on different days, were identical which indicate that the assay is reliable, reproducible and repeatable. The intra-assay and inter-assay coefficient variation was 2.41 (n = 4) and 8.71 (n = 10) respectively. The serial dilution of plasma VTG from exposed tilapia showed a good parallelism with the standard t-VTG within the working range of the assay. The serial dilution of the reference fish did not cover the whole range of the t-VTG standard curve. By using the standard curve and the dilution of the exposed plasma, we were able to demonstrate that the ELISA was able to quantify VTG. With good laboratory practise, this ELISA can be use to quantify VTG in chemically exposed fish. It will also be ideal to continue analyzing the antibody to determine the appropriate dilutions necessary to ensure that the assay work its optimal capabilities. / Dr. I. Barnhoorn Prof. P. Jagals Prof. J.H.J. Van Vuren

The purpose of this research was to investigate the effects of wetland restoration on the water balance, flushing time, and water chemistry of southern Taylor Slough, a major water way in Everglades National Park. Water balance and flushing time equations were calculated on a monthly time step from 2001 – 2011. Water chemistry of major ions and nutrients were analyzed and correlated with water flushing times. Results showed that evapotranspiration followed by water volume had the greatest influence on flushing time. The flushing times varied between 3 and 78 days, with longer times observed between October and December, and the shorter times between March and May. Ion concentrations at the coastal areas decreased with increased flushing times. Increased surface water inflow that resulted from restoration projects and water management changes were productive in the rainy season and should result in increased flushing times and decreased ion concentrations in Taylor Slough.

Water is one of Earth's most valuable resources and one of Earth's most threatened resources. Continuously increasing population growth coupled with changing climate has resulted in the depletion of water sources. As a result, investigations into alternative water sources are being conducted worldwide. One such alternative water source is groundwater abstraction. Groundwater abstraction involves the abstraction of water from an underground source. The volume of water that can be sustainably abstracted is governed by legislation. Groundwater typically requires treatment before it can be distributed to the general population for use, and thus the implementation of large-scale groundwater abstraction projects involves large capital outlays, as well as monthly operational outlays. The feasibility into the implementation of large-scale groundwater abstraction projects is therefore of interest to stakeholders involved in the water supply industry. The lifecycle of a recently implemented large-scale groundwater abstraction project was analysed in order to determine its feasibility. The project was implemented by Drakenstein Municipality in the Western Cape in 2017. The project involved identifying groundwater abstraction points that could provide sustainable volumes of water. The water quality of each groundwater abstraction point was then investigated for any outlying parameters according to SANS 241-1:2015 guidelines for potable water. Groundwater abstraction water treatment plants were then designed in order to treat the combined sustainable flow rates of water at their specific water qualities. The treated water from each groundwater abstraction water treatment plant was then analysed in order to confirm compliance with the SANS 241- 1:2015 guidelines, before the booster pumps were commissioned and commenced with their continuous supply of potable water into the network. The capital expenditure associated with each of the groundwater abstraction water treatment plants was obtained from the Engineer, Aurecon. In addition, the estimated monthly operational expenditure was computed. These expenditures were used to determine the feasibility of the large-scale groundwater abstraction project by computing the payback period and comparing this period to the design life of each of the groundwater abstraction water treatment plants. In addition, the monthly savings applicable to the municipality as a result of the project's implementation was computed. Finally, the feasibility into varying flow rates of groundwater abstraction water treatment plants, and varying water quality of groundwater abstraction points was investigated. Two sites were identified within the municipal area, each with four groundwater abstraction points capable of delivering a combined 5.18 ML/day and 1.62 ML/day. These sites were identified as Boy Louw Sportsgrounds and Parys Sportsgrounds respectively. Although the sites were only 2.60 kilometres apart, the water quality of the combined flow rates indicated that the groundwater abstraction points were accessing two different water sources. The combined sustainable flow rate at Boy Louw Sportsgrounds required turbidity, iron and manganese removal, as well as disinfection. The combined sustainable flow rate at Parys Sportsgrounds required turbidity removal and disinfection. Groundwater abstraction water treatment plants were then designed to treat the water at Boy Louw Sportsgrounds and Parys Sportsgrounds. Boy Louw Sportsgrounds involved the distribution of equipment across seven shipping containers, whilst Parys Sportsgrounds involved the distribution of equipment across three shipping containers. It was found that the groundwater abstraction project was feasible with a payback period of three years. This payback period fell well within the 10-year design life of each groundwater abstraction water treatment plant. In addition, it was found that the municipality would be subject to a 72% monthly saving in water costs as a result of utilising the groundwater abstraction water treatment plants, as opposed to purchasing water in bulk from the City of Cape Town. It was found that the payback periods of Boy Louw Sportsgrounds and Parys Sportsgrounds were two and five years respectively. Although Boy Louw Sportsgrounds delivered almost three times the potable water flow rate than that of Parys Sportsgrounds, its payback period was three years sooner. In addition, it was found that the municipal savings as a result of Boy Louw Sportsgrounds was 8% more than that of Parys Sportsgrounds. It was therefore concluded that the larger the flow rate of water to be treated, the more financially feasible the project. In addition, it was determined that the more water quality parameters lying above the upper limits of SANS 241-1:2015 guidelines for potable water, the more treatment processes would need to be implemented resulting in additional capital and operational expenditure. It was therefore concluded that the more water quality parameters requiring treatment, the less financially feasible the project. Finally, it was determined that the feasibility of the large-scale groundwater abstraction project is limited by the rate at which the municipality purchases water in bulk from the City of Cape Town. As long as the bulk water purchase tariff remains above R 2.85/m³, the project will remain feasible. Should the bulk water purchase tariff fall below this value, the project no longer remains feasible as the payback period of the project exceeds the design life of the groundwater abstraction water treatment plants.

The Prairie Pothole Region, specifically eastern North Dakota, has experienced intense disturbance from agricultural demands and urban sprawl. This study assessed wetlands across the rural, peri-urban, and urban gradient to determine the impacts of urbanization on water quality and vegetation composition. Thirty wetlands were randomly selected and compared based on land use type and the impervious to pervious surface ratio within one mile of each wetland. Water quality samples were taken in 2015 and 2016, and a vegetation assessment was completed at all wetlands. Results indicate disturbance from urbanization impacts wetland water quality and vegetation composition. Rural wetland water quality and vegetation significantly differ from both peri-urban and urban wetlands, whereas peri-urban and urban wetland water quality and vegetation do not differ. Information from this study is useful to wetland professionals across the globe as urban development and sprawl continue to impact wetlands.

Stroubles Creek is registered on Virginiaâ s 303(d) list of impaired waters for both benthic and fecal coliform impairments. The upper reach of the creekâ s watershed drains into two ponds on the Virginia Tech campus. The area draining to the ponds, approximately 715 acres, encompasses most of the Town of Blacksburg and the Virginia Tech campus. Below the ponds, the creekâ s watershed is primarily forested and agricultural, with some areas of residential development. In order to improve water quality downstream, the two ponds will be converted to a water quality facility by redirecting all flow from the northern branch of Stroubles Creek into the upper, smaller pond, which then flows into the larger pond below. With flow into the upper pond increasing dramatically, the dam between the two ponds and associated overflow structures were evaluated and redesigned to protect the dam from overtopping and possible washout. In addition, concrete weirs were designed and will be constructed on both branches of Stroubles Creek above the ponds for future installation of flow and water quality monitoring equipment. Above the ponds, the banks along both branches of the creek have become severely eroded. Interlocking concrete block armoring was designed for the stream banks to reduce erosion and protect the trees growing along the creek. This project was jointly funded by Virginia Tech and a grant from the Virginia Department of Conservation and Recreation Water Quality Improvement Fund. Construction will be performed by the Capital Design department of Virginia Tech. / Master of Science