Thesis (MSc. Agriculture (Soil Science)) -- University of Limpopo, 2019 / Global shortage of fresh quality water has led to the use of treated wastewater in arid and semi-arid regions. Although, the treated wastewater has proven to be the best solution in ameliorating pressures brought by water shortage, it contains toxic heavy metals, some in high concentrations that could possibly pose health risks and degrade soil quality. Therefore, the objectives of the study were to determine the vertical and horizontal distribution of bioavailable heavy metals on virgin, cultivated and fallowed fields and to investigate the bioremediation abilities of selected soil microbes on non-essential heavy metals in cultivated and fallowed soils following irrigation with treated wastewater at University of Limpopo (UL) Experimental Farm. Three fields, namely, virgin field (VF), cultivated field (CF) and fallowed field (FF), each being 6.4 ha, were each divided into 40 equal grids, equivalent to 40 m × 40 m, which were used in vertical assessment of heavy metals. Soil profiles were established inside each grid and soil samples collected at 0-20; 20-40 and 40-60 cm soil depth for further laboratory analysis. The soil samples were analyzed for basic soil physico-chemicals, namely, particle size distribution, soil pH (H20 and KCl), electrical conductivity (EC), reduction potential (Eh), organic carbon (OC) and cation exchange capacity (CEC). Five essential heavy metals namely zinc (Zn), iron (Fe), copper (Cu), cobalt (Co), manganese (Mn) and five non-essential heavy metals, namely, arsenic (As), chromium (Cr), lead (Pb), aluminium (Al), and cadmium (Cd), were also extracted from the soil samples. Heavy metal resistant Gram-negative (–) and Gram-positive (+) bacteria were isolated from the soil and identified as Providencia rettgeri (–), Enterobacter cloacae (–), Bacillus cereus (+) and Arthrobacter aurescens (+). xix The isolated bacteria were cultured and inoculated in heavy metal-contaminated soils and incubated for 12 weeks to bioremediate the non-essential heavy metals. Results obtained suggested that the treatments had no significant (P ≤ 0.05) effects on vertical distribution of all the essential and non-essential heavy metals among the three fields. However, on average Co was above the permissible level at 53 mg/kg in CF at 0-20 cm and although all the other essential heavy metals increased, they were still within the permissible levels. The concentration of As was also above the permissible levels in CF with an average concentration of 4.30 mg/kg. Cadmium levels were also above the permissible levels in CF with an average concentration of 1.146 mg/kg in CF and this increased by 0.46 units from VF which had an average value of 1 mg/kg. However, fallowing reduced Cd to 0.51 mg/kg which was below or within the expected limits in soil previously irrigated with treated waste water. Gram-positive bacteria reduced more concentrations of non-essential heavy metals separately and combined, especially in the fallowed field. Irrigation with treated wastewater has shown to have both negative and positive effects on the concentration of essential and non-essential heavy metals in cultivated and fallowed fields. Bioremediation coupled with fallowing has been proven to be the best solution in ameliorating heavy metal toxicity while naturally improving the quality of the soil. / National Research Foundation (NRF)

Mining industry has been identified as the main sustenance of the South African economy, however the negative impacts of the industry on the ecological systems cannot be over emphasized due to the released waste which is mostly heavy metals into the environment. The study evaluated six heavy metal (A1, Cu, Fe, Ni, Pb and Cr) contents in a tailings dam from a specific mine site. Two sets of samples for the investigation were measured, that is, one in year 2012 and the other in year 2013. In the year 2012, the sample set was only taken at a distance profile of 500 meters from the foot of the dam, whereas the sample set taken in the year 2013 was for the 500 and 1500 meter distance profiles from the foot of the dam. The year 2012 and 2013 sample sets for the 500m distance profile were sampled very similarly to each other. A kilogram of each sample was taken as per grid format. The samples at varied depths were taken at 0-cm depth for the top layer, 20cm depth for the second layer, and 30cm depth for the third layer. The samples for the surface varied distance were taken at 1 m, 2m, 3m, and 4m away from each 500m and 1500 sampling points. The 2012 samples were analysed using characterization methods namely ICP MS and The 2013 samples were analysed using the ICP OES. The comparison of the field results for the six heavy metals studied (A1, Fe, Pb, Cu, Ni and Cr) was performed using statistical analytical methods, namely ANOVA. The statistical analysis results for heavy metals (A1, Fe, Pb, Cu, Ni and Cr) from sample and 2013 revealed that the group means are not significantly different from each other which means that there is no significant difference in (A1, Fe, Pb, Cu, Ni and Cr) concentrations with respect to both depth and distance. The observations from both 2012 and 2013 indicate the results of the samples are in agreement. In addition, the comparative average concentrations of the three results obtained reach the same conclusion that the tailing dam probably does not introduce considerable or significant amounts of these metals (A1, Fe, Pb, Cu, Ni and Cr) into the surrounding soils. / Chemical Engineering / M. Tech. (Chemical Engineering)

Thesis (MScAgric (Soil Science))--University of Stellenbosch, 2007. / Two–thirds of South Africa, including more than 280 towns and settlements are largely dependent on groundwater for their drinking water supply and development. However, groundwater resources in South Africa are limited both in terms of quantity and quality, especially in the semi–arid parts of the country (Sililo et al., 2001, p. i). Therefore, the importance of protecting groundwater resources from pollution has been recognized. The first objective of this research was to investigate the attenuation capacity of a selection of soil horizons and materials representing major types of diagnostic horizons and materials in the South African soil classification in order to validate their chemical attenuation ratings as provisionally specified by Sililo et al. (2001, p. 4.6). The second objective was to assess the pollutant attenuation capacity of South African soil horizons and materials as well as describe the diagnostic value of key chemical properties of soils for conveying information on their contaminant transport/attenuation potential. The third objective was to investigate whether it is possible to apply acid/base priming using H2SO4 and Ca(OH)2 to a bulk quantity of soil in order to reduce the mobility of contaminants.

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

published_or_final_version / Environmental Management / Master / Master of Science in Environmental Management

In the first phase of the study, the effect of five Safol surfactants on the aqueous solubility of phenanthrene and acenaphthene was determined. The fixed variables were temperature and ionic strength, while surfactant concentration and pH were varied. Quantification of the polyaromatic hydrocarbons (PAHs) was conducted by UV-Visible spectrophotometry. The surfactants had little or no effect on analyte solubilisation below the critical micelle concentration (CMC) while a linear relationship between surfactant concentration and amount of solubilised phenanthrene was observed above CMC concentrations. Safol 45E5 had the highest phenanthrene molar solubilisation ratio (0.83) of the five surfactants tested. The solubilisation of phenanthrene increased marginally (4.1 % for Safol 45E12 and 15.2 % for Safol 45E7) by decreasing the pH from 8 to 5. The concentration of solubilised acenaphthene was 8.4 % higher than phenanthrene in a 1 mM solution of Safol 45E7. The aqueous solubility of phenanthrene was enhanced 11.0, 21.2, 19.6, 15.9 and 14.7 times in 1 mM solutions of Safol 45E3, 45E5, 45E7, 45E9 and 45E12 respectively. Seasand, Longlands sand, Longlands soil and a standard soil sample were spiked with the two PAHs and aged for two weeks. API sludge provided by Sasol and unspiked samples of the above mentioned sorbents were subjected to determinations of organic matter content, particle size distribution and moisture content. The spiked soils and sands and the sludge samples were then washed in various concentrations of Safol 45E7 (0.5, 1.0 and 2.0 mM) at the same temperature used in the solubility studies. A soil mass to solution volume of lg to 10 mL was used. Analyses of the soil and sand samples were conducted by High Pressure Liquid Chromatography (HPLC). Using a 2 mM Safol 45E7 surfactant solution, 100 % and 90 % of phenanthrene and acenaphthene were respectively extracted from Longlands sand and 88 % and 100 % of phenanthrene and acenaphthene were removed from seasand. 8.4 % phenanthrene and 8.17 % of acenaphthene was removed from Longlands soil, while 7.03 % phenanthrene and 6.64 % acenaphthene was removed from the standard soil sample. In the sand desorption studies, the amount of desorbed contaminants initially increased rapidly with increasing surfactant concentration, before levelling off at equilibrium. The amount of desorbed acenaphthene and phenanthrene increased exponentially with increasing surfactant concentration while contaminant concentrations decreased with increasing time in the Longlands soil and standard soil desorption experiments. Dry API sludge samples were also subjected to soil washing studies. The washed samples were Soxhlet extracted and analysed by gas chromatography. The 0.5 mM and 1 mM Safol 45E7 washed sludge samples showed respective phenanthrene peak area percent reductions representing a 44 % and 47 % extraction of phenanthrene from the API sludge. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Bacillus cereus is traditionally thought to be the only member of its genus accepted as a pathogen in foods like grains, fruits, vegetables and milk due to the presence of the nonhemolytic (Nhe) operon. However, many other Bacillus spp. may also harbor the Nhe operon and be pathogenic. Real-time PCR targeted the nheA gene in 37 samples obtained from food, soil, and reference cultures by analyzing the standard deviations of melt peaks. Rep-PCR was used to compare the banding patterns of each sample against B. cereus ATCC14579 and three B. thuringiensis strains to “fingerprint” each isolate. Of the original 43 isolated tested, 37 were Gram-positive rods. The remaining six samples were Gram-positive cocci. Twenty-five of the 37 Gram-positive Bacillus spp. were nheA positive, while twelve were negative. Many of the nheA positive strains were species not previously known to contain Nhe, and were capable of causing gastroenteritis in consumers. / Department of Biology

Mining industry has been identified as the main sustenance of the South African economy, however the negative impacts of the industry on the ecological systems cannot be over emphasized due to the released waste which is mostly heavy metals into the environment. The study evaluated six heavy metal (A1, Cu, Fe, Ni, Pb and Cr) contents in a tailings dam from a specific mine site. Two sets of samples for the investigation were measured, that is, one in year 2012 and the other in year 2013. In the year 2012, the sample set was only taken at a distance profile of 500 meters from the foot of the dam, whereas the sample set taken in the year 2013 was for the 500 and 1500 meter distance profiles from the foot of the dam. The year 2012 and 2013 sample sets for the 500m distance profile were sampled very similarly to each other. A kilogram of each sample was taken as per grid format. The samples at varied depths were taken at 0-cm depth for the top layer, 20cm depth for the second layer, and 30cm depth for the third layer. The samples for the surface varied distance were taken at 1 m, 2m, 3m, and 4m away from each 500m and 1500 sampling points. The 2012 samples were analysed using characterization methods namely ICP MS and The 2013 samples were analysed using the ICP OES. The comparison of the field results for the six heavy metals studied (A1, Fe, Pb, Cu, Ni and Cr) was performed using statistical analytical methods, namely ANOVA. The statistical analysis results for heavy metals (A1, Fe, Pb, Cu, Ni and Cr) from sample and 2013 revealed that the group means are not significantly different from each other which means that there is no significant difference in (A1, Fe, Pb, Cu, Ni and Cr) concentrations with respect to both depth and distance. The observations from both 2012 and 2013 indicate the results of the samples are in agreement. In addition, the comparative average concentrations of the three results obtained reach the same conclusion that the tailing dam probably does not introduce considerable or significant amounts of these metals (A1, Fe, Pb, Cu, Ni and Cr) into the surrounding soils. / Chemical Engineering / M. Tech. (Chemical Engineering)

Com a finalidade de estudar o comportamento de fluidos aquosos não miscíveis em água no solo, foram executados ensaios de laboratório, a construção de um modelo físico e a comparação dos dados com a simulação numérica com o programa HSSM. Os ensaios de laboratório tiveram a finalidade de obter os parâmetros do solo utilizado no modelo físico e numérico. Foram obtidas curvas de retenção do solo com concentrações de 2, 5, 10 e 15% de óleo e somente com água, para comparar o comportamento das curvas. Para o modelo físico, foi construído um tanque experimental para simular o derramamento de óleo no solo e, assim, obter uma pluma de contaminação de óleo de maneira controlada. Foram executados três ensaios com o tanque experimental, primeiro com o solo na umidade higroscópica, o segundo com um nível dágua definido, e o terceiro com o solo úmido, mas sem um nível dágua. Destes ensaios no tanque, foram coletadas amostras para a análise em laboratório das concentrações de óleo de cada parte do tanque. A partir dos resultados das curvas características foi possível observar que o óleo influencia principalmente na umidade residual. E a análise das amostras coletadas do tanque experimental indica que a concentração de óleo na pluma estava constante, com concentração de 2% de óleo. Juntando os dados colhidos dos ensaios de laboratório e do tanque experimental, foram executadas simulações da evolução da pluma de contaminação de NAPL para os três casos simulados no tanque experimental. A simulação numérica foi coerente com o modelo físico, mas foi observado que subestima a velocidade de expansão da pluma, principalmente quando o solo está com a umidade baixa. / In order to study the behavior of non-aqueous phased liquids in the soil, laboratory tests were performed, and the construction of a physical model and comparison of data obtained with the numerical simulation with the program HSSM. Laboratory tests are designed to obtain the soil parameters used in the physical and numerical model. Retention curves of soil with concentrations of 2, 5, 10 and 15% of oil and water only were obtained to compare the curves. For the physical model, an experimental tank was built to simulate the oil spill on the ground and have a contamination plume of oil with controlled conditions. Were performed three experiments with the tank, first with the hygroscopic soil moisture, the second with a defined water table, and the third with natural soil moisture, but without a water table. From these tests in the tank, samples were collected for laboratory analysis of concentrations of each part of the oil tank. From the results of the retention curves it was observed that the major oil influence was on residual moisture. And the analysis of samples collected from the experimental tank indicates that the oil concentration in the plume was constant at the concentration of 2% of oil. Combining the data collected in laboratory testing and experimental tank, numerical simulations were performed of the evolution of NAPL contamination plume for the three cases simulated in the experimental tank. The numerical simulation was consistent with the physical model, but it was observed that underestimates the rate of expansion of the contamination plume, especially when the moisture of soil is low.

Hexaclorociclohexano (HCH), pesticida organoclorado mundialmente utilizado, apresenta efeitos tóxicos à saúde humana e ao meio ambiente. Os microrganismos degradadores mais conhecidos são as Sphingomonas sp. Técnicas de biodegradação foram aplicadas em duas etapas. A primeira focou na biorremediação de solo contaminado, de Santo André SP, e foi realizada em biorreatores no Instituto de Pesquisas Tecnológicas (IPT). Experimentos nas fases sólida e semi-sólida apresentaram até 90% de degradação de HCH no solo. A segunda parte, na Universidade de Groningen (RuG), Países Baixos, focou no tratamento de soluções contaminadas usando enzimas selvagens e variantes desenhadas computacionalmente. Mutantes foram construídas, expressadas e purificadas. Ensaios de Thermofluor® mostraram que as variantes estavam enoveladas. Ensaios enzimáticos foram realizados em solução aquosa com b-HCH e dimetil sulfóxido (5%), sendo as amostras extraídas com acetato de etila e analisadas por cromatografia gasosa com detector de captura de elétrons. As variantes apresentaram atividade. / Hexachlorocyclohexane (HCH) is an organochlorine pesticide used world-wide which shows toxic effects in human health and causes environmental problems. The most known HCH-degrading microorganisms are Sphingomonas sp. Biodegradation techniques were applied in this work, divided in two parts. The first one focused on the bioremediation of a contaminated soil, from Santo Andre - SP, in bioreactors at the Institute for Technological Research (IPT). Experiments were carried in solid and slurry phases, which could achieve around 90% of HCH degradation. The second part was developed at the University of Groningen (Rug), The Netherlands. Contaminated solutions were treated with wild-type enzymes and computationally designed variants. Mutants were constructed, expressed and purified. Thermofluor® assay showed that all variants were well folded. Enzymatic assays were carried in aqueous solution with b-HCH and dimethyl sulfoxide (5%). The samples were extracted with ethyl acetate and analysed by gas chromatography using an electron capture detector. The variants were actives.