A total of 34 sewage sludge samples were collected from South African provinces: KwaZulu- Natal, Gauteng, North West and Western Cape province. The samples were analysed for mineral ions and polynuclear aromatic hydrocarbons (PAH). The methods used for sample preparation were US-EPA 3050B and 3540C for mineral ions and PAHs respectively. The moisture content was also analysed. A minimum of 4.5 % and maximum of 99.5 % moisture were detected in the samples. The sludge that had stayed longer in their sampling points had low moisture contents than those that were sampled immediately after production. A total of 22-minaral ions were detected in the samples. When these were quantified an abundance trend that was found to be most common was: P>Ca>Fe>Al>Mg>K>Zn>Na>Mn>Cu>Ba>Cr>Pb>Si>Se>Sr>B>Ni>Mo>Co>Cd>Be. Phosphorous was the most abundant mineral ion in most of the samples apart from calcium, which was found to be abundant in a few cases. Comparing these results to the South African guideline limits, it emerges that most of the sewage works are compliant with the guidelines. The mineral ion that was found to be above the set limit in most occasions was selenium. The current South African guidelines were found to compare favourably to those of other countries. In the study 16 PAHs were also analysed, based on frequency of occurrence these were found to be: benzo(b)fluorathene ~ benzo(k fIuoranthene> phenanthrene> acenaphthylene> fluorine> naphthalene> benzo(a)anthracene> chrysene> acenaphthene> benzo(a)pyrene> fluoranthene> anthracene ~ pyrene> benzo(g,h,i)perylene> indeno(l ,2,3-cd)pyrene> dibenzo(a,h)anthracene Dibenzo(a,h)pyrene was below the detection limit in all the samples analysed while benzo(b)fluoranthene and benzo(k)fluoranthene were the most abundant compounds. All the samples analysed had concentration of benzo(a)pyrene below the South African limit of 2.53 mg kg"1. Comparing these guidelines with other international countries, South Africa specifies a single compound for monitoring while in other countries more than one compound is monitored. Thus South African guidelines need to be put in line with those of major EU countries such as Germany and Denmark with respect to PAH limits. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

A comprehensive literature survey looking at different aspects of organic contaminants in sewage sludge is incorporated in the thesis. It is clear from the data available that plants do not take up most of the organic pollutants. However, a risk of contamination of the food chain exists when sludge is spread directly onto crops that are to be consumed raw or semi-cooked. The major source of human exposure to sludge-borne organic pollutants is through the consumption of animal products such as meat and milk through the bioaccumulation of compounds such as polychlorinated dibenzo-p-dioxins and -furans (PCDD/Fs), polychlorinated biphenyls (PCBs) or polynuclear aromatic hydrocarbons (PAHs). This is due to the ingestion of soil and sludge by livestock due to the spreading of sludge on the land used for grazing. Currently little is known about the plant uptake of phthalates and nonylphenols, which are present in relatively high levels in sludge. Included in the literature survey is a list of the most common methods normally used for extraction of organic compounds and the possible methods of analysis. What is noted is that there is no universally accepted and validated analytical method for analysing most of the organic compounds. In addition, data concerning levels of organic pollutants is scarce worldwide. The methods selected for this project were EPA Methods 35IOC (Liquid-liquid extraction) and 3540C (Soxhlet extraction) for the aqueous and solid sewage sludge respectively. These two methods were chosen because they are simple, inexpensive and effective. The chosen purification method was the sulfur clean-up process (EPA Method 3660B). This is because sulfur precipitates were observed in most of the concentrated extracts, especially from the solid samples. A total of 109 samples from 78 sewage works were extracted, using Soxhlet extraction for solids and liquid-liquid extraction for liquid samples. All the extracts were analysed using GC-MS. The identification of the organic compounds was made possible by the use of GC-MS Wiley library. A total of 712 organic compounds were identified in the South African sewage sludge. These included Phenols, Pesticides, PAHs, Phthalates, PCBs, Furans, Amines, Aldehydes, Esters, Acids, Chlorinated Hydrocarbons, Alcohols, Hydrocarbons and Others (all sorts of organic compounds that did not belong to the named categories). The extraction efficiency and reproducibility using a "clean" sludge matrix was carried out. The results showed that the method chosen for extraction i.e. Soxhlet method had an efficiency of over 80% and the results were reproducible. A total of 14 samples were selected for the quantification of p-cresol, nonylphenol and pesticides listed in the current legislation. The results showed negative results for pesticides, confirming that the pesticides listed in the legislation are not the compounds that need to be regulated as far as organic pollutants are concerned. The reasons for this are most likely due to the fact that most of the pesticides listed are banned or their use is severely restricted. The results for the quantification of p-cresol and nonylphenol, when comparing the liquid and the solid extracts of the liquid sludge, show that 99% of p-cresol is concentrated in the liquid phase. The opposite is true when looking at nonylphenol where 90% of it is trapped in the solid matrix leaving 10% in the liquid phase. It is also noted that liquid sludge contains high concentrations of both p-cresol and nonylphenol when compared to the solid sludge. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Wastewater treatment facilities have been identified as potential source of surface water pollution worldwide. In this study, the physicochemical qualities and heavy metal (including Zinc, Cadmium, Iron, Lead and Copper) concentrations in the final effluents, as well as the reduction efficiencies of three municipal wastewater plants in the Eastern Cape, South Africa were evaluated from September 2015 to February 2016 using standard methods. Wastewater effluent and sludge samples were collected from the selected plants on monthly basis over a period of six months from September 2015 to February 2016. Standard analytical method (Atomic absorption spectroscopy [AAS]) was used to assess the concentration levels of heavy metals. The physicochemical parameters assessed include pH, Temperature, Total Dissolved Solid, Turbidity, Biochemical Oxygen Demand, Chemical Oxygen Demand, Alkalinity, Dissolved Oxygen, Free chlorine, Chloride, Sulphate, Phosphate, Ammonium and Electrical Conductivity. The results of the evaluation showed that temperature ranges from 19 to 36 °C, Electrical Conductivity from 60-1095 mS/m, Alkalinity from 2.6-20.9 mg/L, Nitrate from 0.24-26.5 mg/L, Nitrite from 0.01-90 mg/L, Phosphate from 0.02-5.12 mg/L, Ammonium from 0.06-112 mg/L, Sulphate from 3-72 mg/L, Chloride from 3.25-224 mg/L, Chemical Oxygen Demand from 17-394 mg/L and Turbidity from 1.96-715 NTU. Free chlorine concentrations and dissolved oxygen were within the recommended limits for most part of the sampling period and ranged between 0.17-0.48 mg/L and 0.19-21.9 mg/L respectively. The pH, Phosphate, Sulphate and Free chlorine were within acceptable limits while Electrical Conductivity, Temperature, Total Dissolved Solid, Turbidity, Chemical Oxygen Demand, Chloride, Ammonium, Nitrate and Nitrite were not within specified limits. The reduction efficiencies observed generally ranged between 43.9 and 100 percent at all sampling points while zinc and lead were not detected throughout the sampling period. Cadmium showed low reduction rate ranging from 0 to 11.7 percent, while the rate of Copper reduction ranged from 0 to25 percent in January and February 2016. Zinc, lead and cadmium were generally not detected in the sludge samples except in January and February which could be due to the prevailing atmospheric conditions during the sampling period. Cu and Fe were detected through the sampling months of the study. We conclude that these municipal sewage plants are sources of pollution to their receiving watersheds and threats to public and environmental health.

This research aimed to firstly confirm the hypothesis that the current management of sewage sludge generated during the treatment of wastewater in the Eastern Cape Province of South Africa is poor and, as such, this material poses a threat to local environmental and human health. Secondly, through a rational design approach, the current research also aimed to design and assess the suitability of a novel integrated process incorporating appropriate technologies for improved sewage sludge management. The lack of an appropriate tool for the assessment of the integrated process necessitated the development of an appropriate technology assessment (TA) methodology based on environmental, technical, social and economic criteria. A combination of informal participatory methods such as semi-structured interviews and formal research methods including questionnaires, risk assessment exercises and laboratory analyses were used. Based on the above it was estimated that 116 tons dry sludge were generated in the province per day and that the concentration of heavy metals present in the sludge was generally within the limits for reuse on agricultural land. Furthermore the sludge from all sample sewage treatment works (STWs) was found to be free of any detectable pathogens. Despite the above, a preliminary risk assessment and chemical analysis revealed that existing sludge management practices at sample STWs posed a threat to the environment, particularly to water resources and exacerbated the problems associated with the discharge of poorly treated municipal wastewater. The root causes of the poor sludge management were considered to be a lack of non-regulatory incentives and financial resources. Highly integrated ecologically engineered systems were thought to offer a solution to the dual problem of poor sludge management and municipal effluent treatment, while providing necessary economic incentives. To facilitate the design of a system appropriate to local conditions, it was necessary to first develop a rational design methodology, which incorporated a detailed TA step. The result of the design process was an Integrated Waste Beneficiation Facility (IWBF) that incorporated a number of process units comprised of appropriate technologies including composting, vermicomposting, algal ponding technology and aquaculture. A detailed TA indicated that the benefits of the proposed IWBF would, at the majority of sample sites, outweigh the potential negative impacts and it was thus recommended that investigations should continue on pilot-scale facilities. Furthermore, although the proposed TA based on four sustainability criteria categories was thought to provide a more accurate assessment of the true sustainability of a technology, the acquisition of information was problematic highlighting the need to re-think current TA methodologies and to address associated constraints allowing the tool to be used and fully comprehensive.

M.Sc. (Chemistry) / Prosperity for South Africa depends on the sound management and utilization of many resources, with water playing a crucial role. Located largely in a semi-arid part of the world, South Africa’s water resources are, in global terms, scarce and extremely limited. A key environmental problem facing South Africa is water pollution. This arises from many sources, including mining and industrial effluents, and runoff of biocides, nutrients and pathogens from agricultural lands, urban areas and informal settlements with poor sanitation. The consequences are often severe, including among other impacts, habitat destruction, reduced oxygen levels, fish kills and loss of human life. Inorganic and organic pollutants as well as microbes are the main constituents of the effluent from the domestic, mining, agriculture, metal electroplating, petrochemical and transport industries. The presence of pollutants in environmental systems is of concern because ultimately, they are incorporated into drinking water and various food chains. Therefore, the overall focus of this study was mainly metals analysis in wastewater systems before and after treatment processes so as to establish the efficiency of the treatment processes....

PhD (Environmental Sciences) / Department of Hydrology and Water Resources / See the attached abstract below

Submitted in complete fulfillment for the degree of Doctor of Philosophy (Biotechnology), Durban University of Technology, Durban, South Africa, 2016. / Seasonal nitrification breakdown is a major problem in wastewater treatment plants which makes it difficult for the plant operators to meet discharge limits. The present study focused on understanding the seasonal impact of environmental and operational parameters on nitrifiers and nitrification, in a biological nutrient removal wastewater treatment works situated in the midlands of KwaZulu Natal. Composite sludge samples (from the aeration tank), influent and effluent water samples were collected twice a month for 237 days. A combination of fluorescent in-situ hybridization, polymerase chain reaction (PCR)-clone library, quantitative polymerase chain reaction (qPCR) were employed for characterizing and quantifying the dominant nitrifiers in the plant. In order to have more insight into the activated sludge community structure, pyrosequencing was used in profiling the amoA locus of ammonia oxidizing bacteria (AOB) community whilst Illumina sequencing was used in characterising the plant’s total bacterial community. The nonlinear effect of operating parameters and environmental conditions on nitrification was also investigated using an adaptive neuro-fuzzy inference system (ANFIS), Pearson’s correlation coefficient and quadratic models. The plant operated with higher MLSS of 6157±783 mg/L during the first phase (winter) whilst it was 4728±1282 mg/L in summer. The temperature recorded in the aeration tanks ranged from 14.2oC to 25.1oC during the period. The average ammonia removal during winter was 60.0±18% whereas it was 83±13% during summer and this was found to correlate with temperature (r = 0.7671; P = 0.0008). A significant correlation was also found between the AOB (amoA gene) copy numbers and temperature in the reactors (α= 0.05; P=0.05), with the lowest AOB abundance recorded during winter. Sanger sequencing analysis indicated that the dominant nitrifiers were Nitrosomonas spp. Nitrobacter spp. and Nitrospira spp. Pyrosequencing revealed significant differences in the AOB population which was 6 times higher during summer compared to winter. The AOB sequences related to uncultured bacterium and uncultured AOB also showed an increase of 133% and 360% respectively when the season changed from winter to summer. This study suggests that vast population of novel, ecologically significant AOB species, which remain unexploited, still inhabit the complex activated sludge communities. Based on ANFIS model, AOB increased during summer season, when temperature was 1.4-fold higher than winter (r 0.517, p 0.048), and HRT decreased by 31% as a result of rainfall (r - 0.741, p 0.002). Food: microorganism ratio (F/M) and HRT formed the optimal combination of two inputs affecting the plant’s specific nitrification (qN), and their quadratic equation showed r2-value of 0.50. This study has significantly contributed towards understanding the complex relationship between the microbial population dynamics, wastewater composition and nitrification performance in a full-scale treatment plant situated in the subtropical region. This is the first study applying ANFIS technique to describe the nitrification performance at a full-scale WWTP, subjected to dynamic operational parameters. The study also demonstrated the successful application of ANFIS for determining and ranking the impact of various operating parameters on plant’s nitrification performance, which could not be achieved by the conventional spearman correlation due to the non-linearity of the interactions during wastewater treatment. Moreover, this study also represents the first-time amoA gene targeted pyrosequencing of AOB in a full-scale activated sludge is being done. / D

Water is an essential need that stimulates health and well being. Increase in population size and urbanization negatively affect water resources due to high demands of effluent outputs. Wastewater is an important reservoir for Escherichia coli and can present significant acute toxicity if released into receiving water sources without being adequately treated. E. coli is used as indicator organism for the detection of faecal contamination. These strains have been considered to be one of the primary causes of diarrhoeal infections worldwide. The present study was conducted between September 2012 and June 2013 to assess the prevalence of pathogenic E. coli strains in the final effluents of four wastewater treatment plants in Chris Hani and Buffalo City Municipalities in the Eastern Cape Province of South Africa. Standard membrane filtration technique was used for bacteriological analysis and molecular based technique was used for identification of E. coli pathotypes. The results were recorded in colony forming units/100 ml. Faecal coliforms ranged between 0-9.6×10³ CFU/100 ml for the wwtp-Q and E. coli densities ranged between 0-8.4×10³ CFU/100ml. Faecal coliforms ranged between 4×10²-9.7×10³ CFU/100 ml for wwtp-M and E. coli densities ranged between 1.2×10¹-8.4×10³ CFU/100 ml. The wwtp-E showed to have bacterial counts of faecal coliforms ranging between 4.0×10³-8.2×10³ CFU/100 ml and E. coli densities ranging between 3.5×10¹-7.1×10³ CFU/100 ml. The WWTP-K in this study was only assessed for the presence of E. coli. Faecal coliforms were assessed by the other members of the group. This plant showed to have E. coli densities ranging between 0-7.5×10²CFU/100 ml. A total of 200 presumptive E. coli isolates were subjected to screening by conventional PCR in which (29%) of the wwtp-M isolates were positively identified as E. coli, (16%) of the wwtp-K, (22%) of the wwtp-Q and (34%) of the wwtp-E isolates were positively confirmed as E. coli. A total of 100 randomly selected E. coli isolates were characterised into different pathotypes. (16%) of positive isolates were detected as EPEC and 11% were detected as UPEC strains. There was no detection for the ETEC strains. Antibiotic susceptibility patterns of E. coli strains showed high levels of resistance to Penicillin G, Erythromycin, Tetracycline and Sulfamethoxazole. High levels of Susceptibility were observed in antibiotics such as Chloramphenicol, Amoxicillin and Tetracycline. The results of this study reveal that the plants were above the recommended Standard limit of zero CFU/100 ml for effluents meant to be discharge into receiving water sources. This study reveals inadequacy of the plants studied to produce effluents of acceptable quality.

Wastewater discharges may contain health compromising pathogens and carcinogenic and/or chemical substances that could compromise the public health and impact negatively on the environment. The present study was conducted between August 2007 and July 2008 to evaluate the Listeria abundance (as free-living and plankton associated species) and physicochemical qualities of the final effluents of three wastewater treatment facilities in the Eastern Cape Province of South Africa selected to represent typical urban, peri-urban and rural communities and the impact of the discharged final effluents on their respective receiving watershed, as well as to elucidated the in vitro antibiotic susceptibilities and resistance genes profile of Listeria species isolated from the final effluents. The suitability of the secondary effluent of the urban treatment facility (as a case study) for use in agriculture and aquaculture with reference to recommended standards was also determined. Wastewater samples were collected from the raw sewage, secondary effluent, final treated effluent, discharge point, 500 m upstream discharge point, and 500 m downstream discharge point from all three locations on a monthly basis throughout the study period. Listeria abundance in the final effluents and the receiving watersheds varied between 2.9× 100 and 3.52 × 105cfu/ml across the sampled locations. Free-living listerial density across the sampled locations ranged between 0 and 3.2 × 103cfu/ml while counts of Listeria species attached to large (180 μm) planktons varied from 0 to 1.58 × 105 cfu/ml and those of the 60 and 20 μm categories were in the range of 0 to 1.32 × 103 cfu/ml and 0 to 2.82 × 105 cfu/ml respectively. Listeria abundance did not vary significantly with location and season; there was however, significant (P < 0.05; P < 0.01) variance in Listeria abundance with plankton sizes across the locations. Free-living Listeria species were more abundant in the rural and urban xii communities than plankton attached Listeria species; whereas the reverse was the case in the peri-urban community. Prevalence of Listeria in terms of total counts was 100 percent across all sampled locations. Free-living Listeria species showed prevalence ranging from 84-96 percent across the sampling locations; while Listeria species attached to large (180 μm) planktons exhibited prevalence ranging from 75 percent to 90 percent. The prevalence of medium-sized (60 μm) plankton associated Listeria species varied between 58 percent and 92.5 percent; whereas those of Listeria species attached to small (20 μm) planktons ranged from 65-100 percent across all three communities. Listeria prevalence was generally a reflection of the turbidity of the water system, with free-living Listeria species being more prevalent than plankton associated cells in the relatively less turbid rural and urban waters compared to the more turbid peri-urban waters where plankton attached cells were more prevalent in comparison with their free living counterparts The final treated effluent quality fell short of recommended standards for turbidity, chemical oxygen demand and phosphate across all three communities. In addition, the final effluent of the rural treatment plant also fell short of recommended standard for NO3, while that of the urban treatment plant did not comply with acceptable limits for dissolved oxygen and nitrite. Other physicochemical parameters were compliant with set standards after treatment. An inverse relationship was observed between chlorine residual and listerial density across the sampled facilities; the effect of chlorine was however not enough to eliminate the pathogen from the water systems. At the urban treatment plant and its receiving watershed, pH, temperature, EC, turbidity, TDS, DO, and nitrate varied significantly with season and sampling point (P < 0.05; P < 0.01). Salinity also varied significantly with sampling point (P < 0.01), while COD and nitrite varied significantly with season (P < 0.05). Although, the treated effluent fell within recommended water quality standard for pH, TDS, nitrate and nitrite, it fell short of stipulated standards for other parameters. Whereas the microbial quality of the secondary treated effluent at this (urban) facility fell short of recommended standard after secondary treatment, its physicochemical quality were generally compliant with recommended standards for reuse wastewater in agriculture and aquaculture. Listeria pathogens isolated from effluents of the rural wastewater facility were sensitive to 11 (55 percent) of the 20 test antibiotics, and showed varying (7-71 percent) levels of resistance to 8 antibiotics; whereas those isolated from the peri-urban community showed sensitivity to 6 (30 percent) of the 20 test antibiotics, and varying (6-94 percent) levels of resistance to 12 antibiotics; while the urban effluent isolates were sensitive to 3 (15 percent) of the 20 test antibiotics, and showed varying (4.5-91 percent) levels of resistance to 17 antibiotics. Multiple antibiotic resistances involving 78.5-100 percent of isolates and antibiotics combination ranging from 2-10 antibiotics was observed across the sampled locations. Penicillin G and ampicillin showed remarkably high (64-91 percent) phenotypic resistance across the three sampled facilities. Other antibiotics, to which isolates showed significant resistance, were linezolid (22-88 percent); erythromycin (43-94 percent) and sulphamethoxazole (7-94 percent). Two of the 14 Listeria strains isolated from the rural effluents were positive for ereA and sul1 antibiotic resistance genes; while sulII genes were detected in five of the 23 Listeria isolates from the urban effluent and none was detected in isolates from the peri-urban community. The presence of antimicrobial resistance genes in the isolates did not correlate with phenotypic antibiotic resistance. The current study demonstrated that Listeria pathogens easily survived the activated sludge treatment process as free-living and plankton attached entities and suggests that municipal wastewater treatment plants are a significant source of multiple resistant Listeria pathogens in the South African aquatic milieu. While the physicochemical quality of the urban final effluent suggests that it is a major source of pollution to the receiving watershed, the secondary effluent quality demonstrated a great potential for use in agriculture and aquaculture.

MENVSC / Department of Hydrology and Water Resources / Sewage sludge (biosolids) from wastewater treatment plants (WWTPs) has been widely used as a soil improver in Europe, United States of America and some developing countries including South Africa. It has its benefits for farmers as a good source of organic matter and minerals, however, sludge after treatment still contains pathogenic organisms, heavy metals and persistent organic pollutants (POPs). The POP and heavy metal contaminants that accumulate in sludge may transfer through the food chain and cause adverse effects on human beings. In this study, a field experiment was carried out on farmland fertilized with sewage sludge from a wastewater treatment plant (WWTP) that vasically receives domestic wastewater and storm water. Vegetable spinach (Spinacia oleracea) was used for this study and was planted on a farmland under controlled conditions. Ten ridges each of dimensions 20 m × 0.3 m was made and dry sludge weights of 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 kg were applied as manure on each of the ridges, respectively. Representative samples of sludge and soil were taken for analysis of heavy metals and POPs. At maturity, in twelve weeks, the root and leave samples of the vegetable were taken from all the ridges including the control. The soil, sludge, and vegetable samples were analyzed for total heavy metal content (Cd, Cr, Cu, Ni, Pb, Co, Zn, Al, Fe, Mn), speciated heavy metal content and POP (PAH, PCB). Soil and sludge samples were also analyzed for total organic content, pH, cation exchange capacity (CEC), conductivity and alkalinity. The analysis for total heavy metals and speciated heavy metal content was carried out using inductively coupled plasma optical emission spectrophotometer (ICP-OES), and CEC analysis was carried out using atomic absorption spectrophotometer (AAS). A two-dimensional gas chromatograph with time of flight mass spectrometry detector (GC X GC TOFMS) was used for POP measurements. pH measurement was made using a pH meter and conductivity measurement using a conductivity meter. Alkalinity and total organic content analysis was performed using titrimetric apparatus. The highest total heavy metal concentration of 378.9 mg / kg was recorded in Fe metal in soil and Leaf sample while the lowest total metal concentration of 0.0003 mg / kg was in Cu metal in root sample. The highest heavy metal concentration of 1002 mg / kg in speciated forms was in Mn metal in F1 fraction and the lowest of 0.0004 mg / kg was in Cd metal in F5 fraction. PAHs were only found in soil samples and their concentrations ranged from 2.53 mg / kg to 146.5 mg / kg. There were no PCB detected in all the samples analysed. The results indicated that the trace metals concentrations found in the exchangeable fraction were higher than those observed in any of the preceding extractions except in the case of Cd, Cr, Fe and Pb where Fe-Mn oxide and organic matter fractions predominated and were closely followed by exchangeable fraction.