Thesis (M. Sc.)--University of Hong Kong, 1999. / Includes bibliographical references (leaves 86-88).

Thesis (MSc Food Sc )--Stellenbosch University, 2004. / ENGLISH ABSTRACT: The South African apple juice processing industry is growing rapidly and during the harvesting season the wastewater volumes and organic loads increase significantly with a considerable environmental impact. These larger apple juice processing wastewater (AJPWW) volumes and chemical oxygen demand (COD) loads subsequently lead to faster increases in the organic loading rate (OLR) of an upflow anaerobic sludge bed (UASB) wastewater treatment system and it is necessary to know if the treatment system can handle such drastic increases over short periods. The objective of the study were to evaluate the efficiency of the UASB process in the treatment of an AJPWW; to determine what effect a substrate viscosity increase, based on a pectin calcium gel has on the performance of an UASB system, and to determine what impact ozonation has on the pectin content, gelformation ability and biodegradability of the AJPWW. The ability of the UASB to maintain stability during the apple-processing season was investigated by increasing the OLR from 2.9 to over 14.0 kg COD.m-3.d-1 in 131 days. During this time the COD removal remained constant at 85%, while the pH and alkalinity remained at levels indicative of good reactor stability. It was thus concluded that the UASB reactor could operate successfully during the apple-harvesting season when wastewater volumes and organic loads increase significantly. In the study it was found that the viscosity of the AJPWW, containing 750 mq.L-1 pectin, increased from 8.5 to 47.0 cps after a 312 rnq.L-1 Ca2 + addition. This increased viscosity substrate was then fed to an UASB reactor at an OLR of 15.0 kg COD.m-3.d-1. During a 12 day increased viscosity (47 cps) feeding stage the COD removal decreased from 94 to 11%, while the reactor pH decreased from 7.5 to 4.9. During this period, pectin accumulated in the UASB and led to biomass washout and rapid UASB failure. The possible elimination of pectin by ozonation was thus investigated, and a 77% decrease in pectin content and 76% decrease in gel formation ability occurred after ozonation. The effect of pre- and post-ozonation on the efficiency of the UASB system was subsequently investigated. It was found that a 10 min pre-ozonation decreased the AJPWW COD by 19% and the total suspended content by 36%, while the soluble portion of the total COD was increased from 81.7 to 92.4%. This increase in soluble COD content should lead to increased wastewater biodegradability. The ozonated AJPWW was then used to replaced the raw non-ozonated AJPWW as reactor feed. Results showed that the COD removal increased from 78 to 90% within 24 h of starting with the ozonated feed. It was also found that the reactor stability improved after AJPWW pre-ozonation as an OLR increase from 10.0 to 16.6 kg COD.m-3.d-1 in 23 days did not detrimentally influence the stability of the reactor. This reactor effluent (COD = 465 rnq.L-1) was then post-ozonated which resulted in 64.8% COD and 79.0% colour reductions. The final effluent had a COD of 180 rnq.L-1 (98% reduction). The ability of the ozonation/digestion system as described in this study to degrade AJPWW at a higher OLR is of value to the apple industry, as it may lead to larger organic pollutant removals and thus a more effiecient treatment system. Increased reactor performance will directly improve the quality of the final wastewater produced, which in turn will have a significant impact on the treatment ability of the South African apple processing industry currently limited by the production of large wastewater volumes. / AFRIKAANSE OPSOMMING: Gedurende die oesseisoen word groot volumes afvalwater met 'n hoe organiese lading in die vinnig groeiende Suid-Afrikaanse appelsapprosesseringsbedryf geproduseer. Dit het 'n groot impak op die omgewing. Die groter volumes appelsapprosesseringsafvalwater (ASPAW) met 'n hoë organiese lading het 'n vinniger verhoging in organiese ladingstempo's (OLT) van 'n UASB-waterbehandelingstelsel tot gevolg. Daarom is dit belangrik om te weet of die stelsel die drastiese verhoging oor kort tydperke kan hanteer. Die doel van hierdie studie was die evaluering van die effektiwiteit van die UASB-proses in die behandeling van ASPAW; om te bepaal watter effek 'n substraatviskositeitsverhoging, gebaseer op 'n pektien-kalsium-jel, op die doeltreffendheid van 'n UASB-stelsel het; en om te bepaal watter impak osonering op die pektieninhoud, jelvormingsvermoe en bioafbreekbaarheid van ASPAW het. Die vermoe van die UASB om stabiliteit te handhaaf gedurende die appelsapprosesseringseisoen is ondersoek deur die OLT van 2,9 tot bo 14,0 kg CSB.m-3.d-1 te verhoog oor 131 dae. Gedurende hierdie tyd het die chemiese suurstofbehoefte- (CSB-) verwydering konstant gebly by 85%, terwyl die pH en alkaliniteit ook op vlakke aanduidend van goeie reaktorstabiliteit gebly het. Daar is sodoende bewys dat die UASB-reaktor suksesvol kan presteer tydens die appelsapprosesseringseisoen, wanneer daar 'n beduidende verhoging in OLT plaasvind. In die studie is daar gevind dat die viskositeit van die ASPAW, wat 750 mq.L-1 pektien bevat, van 8,5 tot 47,0 cps toeneem na die byvoeging van 312 rnq.L-1 Ca2+. Hierdie verhoogde vikositeitsubstraat is tot die UASB-reaktor toegevoeg teen 'n OLT van 15,0 kg CSB.m-1.d-1. Gedurende 'n 12-dae toevoer van verhoogde viskositeit (47 cps), het die CSB-verwydering van die reaktor afgeneem van 94% na 11%, terwyl die pH gedaal het van 7,5 na 4,9. Gedurende hierdie tydperk het pektien in die UASB geakkumuleer, wat gelei het tot die uitspoel van biomassa en vinnige UASB-reaktormislukking. Die moontlike eliminasie van pektien, deur osonering, is daarom ondersoek. 'n Verlaging van 77% in pektieninhoud en 76% in jelvormingsvermoe het na osonering plaasgevind. Die effek van pre- en post-osonering op die effektiwiteit van 'n UASB-stelsel is gevolglik ondersoek. Daar is gevind dat 'n 10 minute pre-osonering die CSB van die ASPAW met 19% verlaag en die totale inhoud van gesuspendeerde vaste stowwe met 36% verlaag, terwyl die oplosbare gedeelte van die totale CSB van 81,7% tot 92,4% gestyg het. Die verhoging in oplosbare CSB-inhoud behoort tot verhoogde bioafbreekbaarheid van ASPAW te lei. Die geosoneerde ASPAW is gebruik om die rou, ongeosoneerde ASPAW as reaktorsubstraat te vervang. Die resultate het getoon dat die CSB-verwydering verhoog het van 78% na 90% na 'n 24-uur toevoer van geosoneerde substraat. Daar is ook gevind dat die reaktorstabiliteit toegeneem het na ASPAW osoneering, aangesien 'n OLT-verhoging van 10,0 na 16,6 kg.CSB.m-3.d-1 in 23 dae nie die stabiliteit van die reaktor nadelig beinvloed het nie. Hierdie reaktoruitvloeisel (CSB = 465 rnq.L-1) is hierna gepost-osoneer, wat 'n 64,8% CSB- en 79,0% kleurverlaging tot gevolg gehad. Die finale uitvloeisel het 'n CBS-inhoud van 180 rnq.L-1 gehad (98,1% verwydering). Die vermoe van die osonering-/verteringstelsel om ASPAW te degradeer teen 'n hoër OLT, soos beskryf in hierdie studie, is van waarde tot die appelsapprosesseringsbedryf, aangesien dit tot groter organiese afvalstofverwydering kan lei en dus 'n meer effektiewe behandelingstelsel tot gevolg kan hê. Verhoogde reaktordoeltreffendheid sal 'n direkte verbetering tot gevolg hê in die gehalte van die finale afvalwater wat geproduseer word, wat op sy beurt 'n beduidende impak sal hê op die behandelingsvermoe van die appelsapprosesseringsbedryf, wat tans beperk word deur die produksie van groot volumes afvalwater.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A primeira etapa do presente trabalho consistiu em caracterizar um efluente lácteo industrial (soro de queijo) em termos microbiológicos e físico-químicos visando à obtenção de um consórcio de microrganismos para a sua biodegradação. Foram determinadas a contagem padrão de bactérias heterotróficas e a contagem de fungos filamentosos e leveduras de acordo com as metodologias oficiais, e a caracterização físico-química foi realizada em termos do teor de proteínas, cinzas, lipídios, carboidratos, pH e demanda química de oxigênio (DQO). O consórcio microbiano foi obtido em biorreator operando em batelada e contendo 1,5L do efluente, a 200rpm, aeração de 1vvm e temperatura de 28 - 30oC. Os cultivos foram realizados em duplicata durante onze dias, sendo realizado um pulso de 150mL do efluente (10% do volume útil do biorreator) no sétimo dia do cultivo. Foram obtidas concentrações iniciais de 3x109UFC/mL e 8x104UFC/mL para contagem total de bactérias e de fungos e leveduras, respectivamente. O efluente apresentou uma DQO inicial de 64.000mgO2/L, pH 5,0, e concentrações correspondentes a 5g/100mL de carboidratos, 0,8g/100 mL de proteínas e 0,6g/100 mL de lipídios. Os cultivos do efluente no biorreator demonstraram um aumento da concentração de bactérias ao longo do tempo, atingindo 1x1013UFC/mL após oito dias, enquanto que as concentrações de fungos filamentosos e de leveduras foram inibidas. Foi observada uma redução da DQO de 47 a 63,5% após 11 dias de cultivo, demonstrando a potencialidade da utilização do consórcio microbiano no tratamento do efluente. Ensaios qualitativos demonstraram a presença das enzimas: lipase, amilase, protease e celulase no extrato metabólito do consórcio microbiano. No entanto, foram obtidos baixos valores de atividade lipolítica quando comparados com a atividade da enzima comercial, sendo observado ainda que a atividade decresce ao longo do tempo. Na segunda etapa do trabalho foi avaliada a hidrólise enzimática de material flotado, obtido a partir do processo de flotação em coluna do efluente, utilizando a técnica de planejamento de experimentos. Inicialmente, foi avaliado o desempenho de uma lipase comercial (lipolase), sendo investigados os efeitos da agitação e da concentração de enzima sobre a hidrólise. Foi realizada a análise estatística considerando os efeitos significativos para um nível de confiança de 95%. Concentrações maiores de enzima dentro da faixa investigada (1 a 7%) maximizam a hidrólise de óleos e gorduras do material flotado, bem como para níveis de agitação entre 100 e 150rpm. Em todos os experimentos houve a liberação de ácidos graxos no meio em percentuais próximos a 100% apenas após 4 horas de reação. Vale salientar ainda que percentuais de liberação de ácidos graxos superiores a 92% foram obtidos para toda a faixa de concentração testada. Uma agitação de 150rpm foi utilizada em um segundo planejamento experimental, avaliando-se a concentração da enzima comercial associada a um consórcio microbiano, sendo verificado que a adição do consórcio não influenciou de maneira positiva o aumento do teor de ácidos graxos livres no meio. / The first step of the present work was characterizing a dairy effluent in terms microbiological and physical-chemical used to obtain a microbial consortium for its biodegradation. Microbiological characterization was performed by total heterotrophic bacteria, filamentous fungi and yeasts count according to the official methodologies. Physical-chemical characterization was accomplished by content of protein, ash, lipids, carbohydrates, pH and chemical oxygen demand (COD). Microbial consortium was obtained in batch using a bioreactor containing 1,5L of dairy effluent, at 200 rpm, 1vvm, at 28 - 30oC. Cultivations were performed in duplicate for eleven days, being applied a pulse of 10% (v/v) of effluent on the seventh day of the experiment. Cellular growth kinetics, chemical oxygen demand concentration and lipolytic activity were determined. Initial concentrations of bacteria and fungi and yeasts were equal to 3x109CFU/mL and 8x104CFU/mL, respectively. Effluent presented a COD of 64.000mgO2/L, pH 5,0 and concentrations of 5g/100mL of carbohydrates, 0,8g/100mL of protein and 0,6g/100mL of lipids. Cultivations showed an increase on the bacteria concentration, reaching 1x1013CFU/mL after eight days, while filamentous fungi and yeast concentrations were inhibited. An reduction of COD concentration in the range of 47 to 63,5% was observed after eleven days of cultivation, demonstrating the potentiality of use of microbial consortium for effluent treatment. Qualitative assays showed the presence of the enzymes lipase, amylase, protease and cellulase in the metabolic extract of the microbial consortium. However, low values of lipolytic activity were obtained as compared to the commercial enzyme, being observed that activity decreases along time. In the second step of this study, was evaluated the enzymatic hydrolysis of floated material, obtained from the column flotation of a dairy industry, using the technique of experimental design. Initially, performance was evaluated by a commercial lipase (lipolase), being investigated the effects of agitation and enzyme concentration on hydrolysis. A statistical analysis was performed considering the significance of effects to a confidence level of 95%. Higher enzyme concentrations in the range investigated (1 to 7%) maximize the hydrolysis of oils and fats in the floated material, and to agitation levels between 100 and 150rpm. In all experiments, fatty acids were released in rates near to 100% only after 4 hours of reaction. Moreover, rates of released fatty acids were greater than 92% in all the concentration range tested. An agitation of 150rpm was used in a second experimental design, evaluating the concentration of commercial enzyme associated with a microbial consortium. The kinetic monitoring of the process showed that the addition of the consortium did not influence in a positive manner the increase the content of free fatty acids.

Thesis (MTech (Public Management))--Peninsula Technikon, Cape Town, 2004 / In South Africa, an estimated 15 million people - 38% of our population - do not have adequate sanitation. Every citizen has a constitutional right of access to basic services, which local government has the responsibility to provide. In reality such provision to people living in poverty is a daunting development challenge, exacerbated by growing unemployment and the spread of unplanned informal settlements. On the other hand, increased government investment in accelerating provision is a significant opportunity to link sanitation delivery to local economic development, as suggested in the recently revised Water Services Strategy document (DWAF, 2003). There is evidence that these two aspects of national policy can be brought together fruitfully. Community involvement, described as "a commitment to building on people's energy and creativity" (WSSCC, 2001) is consistently advocated by international, national and local government (DPLG, 2001) as essential to sanitation provision. There is broad agreement that a community-based approach is the cornerstone of sustainable service provision. This research focused on the context of urban poverty in informal settlements, taking community responses to sanitation delivery by local authorities into account. Prevailing approaches have had limited success in preventing health hazards, which relies on community-level actions to deal with poor use, inadequate maintenance and dysfunction of such sanitation services as are provided. Implicit in the principles underlying the involvement of communities are substantial community-based roles and functions that the research seeks to make explicit. Diverse local level capacities emerge as quite distinct opportunities for residents to become more actively involved in improving and sustaining their sanitation services.

Thesis (MTech (Public Management))--Cape Peninsula University of Technology, 2017. / The communities of Khayelitsha face problems with regard to managing waste. Increase on the generation of solid waste in the households by the members of the community; far exceed the townships’ ability and capability for safe disposal. Littering in the township is also a persistent problem, despite various clean–up and anti-litter promotional campaigns and programs introduced by local government, private organizations and other community interest groups. The persistent problem of litter and mismanagement of solid waste in the household should be addressed. The rationale and assumption underpinning this study was that waste management problems are related to social norms and attitude of the people of Khayelitsha. To investigate this phenomenon, the study adopted a qualitative paradigm. 300 heads of household residing in Khayelitsha for more than 5 years were surveyed, and door to door interview questions were conducted in 2016 June. Data from the questionnaire were analysed using Thematic and coding analysis. NVivo software was used to generate frequency tables. The responses to most interview questions were consistent for all categories of respondents and did not vary according to respondents’ background, such as gender, location household size or education. The study showed that social norms and attitudes towards waste minimisation in Khayelitsha are related to the entire waste management operation, and householders in Khayelitsha think similarly. To improve waste management in Khayelitsha the following are recommended: • Government and business need to consider incentives to minimise waste; • Government needs to improve waste management service facilities and build recycling centre which are accessible to the community; and • Government and non-governmental organizations and community members should consider awareness, education and training programs on waste wise management.

Orientador: Bruno Coraucci Filho / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-04T22:16:59Z (GMT). No. of bitstreams: 1 Bellingieri_PauloHenrique_M.pdf: 9365011 bytes, checksum: 6c7e81559cc6178c82c39aab2cd464f1 (MD5) Previous issue date: 2005 / Resumo: As águas subterrâneas representam a principal preocupação no processo de viabilização do reúso agrícola, como prática de pós-tratamento e disposição final de esgotos sanitários. O presente trabalho visa avaliar a possível contaminação gerada pela disposição controlada de efluente de tratamento secundário no cultivo de milho, e identificar a lâmina hídrica ideal para a irrigação da cultura, aliando as necessidades da engenharia sanitária com as da engenharia agrícola. Consistiu na análise de amostras do lençol freático, coletadas periodicamente durante duas saltas consecutivas, sendo uma na estação chuvosa e outra na estação seca. Os parâmetros analisados foram: pH, condutividade elétrica, Cr, Pb, Ni, Cd, Cu, Zn e N-N03-. Uma rede de poços de monitoramento foi instalada na área de pesquisa, contendo 10 poços locados a jusante de cada parcela para avaliação em função do risco à saúde pública. A variação do comportamento do lençol freático foi monitorada pela confecção de mapas potenciométricos sazonais e realização de slug test para obtenção da condutividade hidráulica dos poços de monitoramento. Os resultados permitiram concluir que a aplicação de efluentes sanitários na agricultura, via sulcos rasos, causou alguns impactos negativos às propriedades do solo e água subterrânea, além de proporcionar benefícios para a cultura, quando da reposição de água ao solo em períodos de estiagem / Abstract: Groundwaters represent the main concern in the process of viabilization of agriculture reuse, as practice of post-cure and final disposal of sanitary sewers. The present research aims to evaluate the possible contamination generated for the controlled disposal of the sewer of secondary treatment in the maize culture, and to identify the ideal hydric blade for the irrigation of the maize culture, uniting the necessities of sanitary and agricultural engineering. It consisted in the analysis of samples of the water table, collected periodically during two consecutive harvests, one in the rainy and another in the dry period. Were analyzed the following parameters: pH, electrical conductance, Cr, Pb, Ni, Cd, Cu, Zn and N-NO3 . A net of monitoring wells was installed in the research area, with 10 leased wells the downward of each parcel for evaluation in function of the risk to the public health. The variation of the behavior of the water table was monitored by the confection of potenciometric maps and accomplishment of slug test for attainment by the hydraulic conductivity of the monitoring wells. The results allowed to conclude that the application of sanitary effluent in agriculture, in saw ridges fIat, did not cause significant negative alterations in the properties of the ground and underground water, besides providing benefits for the maize culture, within of the replacement water soil in dry periods / Mestrado / Saneamento e Ambiente / Mestre em Engenharia Civil

Orientadores: Durval Rodrigues de Paula Junior, Sylvio Luis Honório / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Agrícola / Made available in DSpace on 2018-08-23T17:03:06Z (GMT). No. of bitstreams: 1 Aguiar_RosaHelena_D.pdf: 2411611 bytes, checksum: cc0a7bb404d742457c915d8126132bf5 (MD5) Previous issue date: 2013 / Resumo: O saneamento além de ser importante para a preservação dos recursos naturais representa uma ação preventiva eficaz para a melhoria do bem estar e da qualidade de vida da população nas questões relacionadas à saúde pública. A prática do uso de água residuária tem grandes vantagens, sobremaneira no aporte de nutrientes às plantas, fator que contribui para um crescimento mais rápido. Nesta pesquisa optou-se por utilizar o girassol (Helianthus annuus L.) ornamental que foi irrigado com águas residuárias. Objetivou-se a viabilidade do uso da área com disposição de esgoto tratado em reatores anaeróbios, por meio da avaliação do seu desenvolvimento e da sua produtividade durante três diferentes épocas de semeadura, visando reduzir os custos com nutrientes. Realizou-se esse experimento na área experimental da Faculdade de Engenharia Agrícola FEAGRI/UNICAMP, e foram avaliados cinco diferentes experimentos, dois deles por meio de reuso de efluentes de reatores - Upflow Anaerobic Sludge Blanket (UASB) e Reator Anaeróbio Compartimentado (RAC) associado a filtros anaeróbios, enquanto que os outros foram constituídos de: Testemunha, sem qualquer tipo de irrigação; com irrigação, utilizando somente água tratada, e com água tratada em área fertilizada com NPK. Foram avaliados respostas fitomorfológicas da cultura, assim como: diâmetro dos caules (DC); altura das plantas (HP); diâmetro dos capítulos (DCp); massa seca dos caules e das folhas (MS); número de folhas (NF) e sua produtividade. Os sistemas modulares (UASB e RAC) de tratamento de esgoto apresentaram um bom desempenho, com elevados valores de remoção de SSed (98,72; 98,36%), e DQO (72,37; 75,79%) não havendo diferença significativa entre os sistemas de tratamento. Constatou-se também que os canteiros com tratamentos com reuso de efluentes promoveram alteração na fertilidade do solo, com o aumento nos valores de saturação por bases (V%) na profundidade de 0 - 20 cm. Analisando os efeitos sobre a cultura nos três plantios, verificou-se um melhor desenvolvimento nos canteiros com os tratamentos do efluente, quando comparado com os demais tratamentos principalmente no segundo e terceiro plantio. Sistemas modulares de tratamento de esgotos associados ao reuso agrícola de seus efluentes podem agregar valores à produção agrícola, podendo ser utilizado inclusive no período de chuvas e em locais onde há escassez de água pluvial / Abstract: Sanitation is also important for the preservation of natural resources, represents an effective preventive action to improve the well-being and quality of life of the population on issues related to public health. The practice of using wastewater has great advantages, particularly in the supply of nutrients to plants, factor that contributes to a faster growth. In this research we decided to use the Sunflower (Helianthus annuus L.) ornamental that was irrigated with wastewater. The viability of the use of the area with provision of treated sewage in anaerobic reactors, by assessing their development and their productivity during three different periods of sowing to reduce costs with nutrients. This research was developed in the experimental area of the Faculty of agricultural engineering FEAGRI/UNICAMP, and evaluated five different treatments, two of them by means of Upflow Anaerobic Sludge Blanket reactors (UASB) and Anaerobic Reactor Chambered (ARC) associated with anaerobic filters, while the others were made up of: witness, without any kind of irrigation; with irrigation, using only treated water and treated water in the area fertilized with NPK. Fitomorfológicas culture responses were evaluated as: diameter of stems (DS); plant height (HP); diameter of the chapters (DCp); dry mass of stems and leaves (DM); number of leaves (NF) and your productivity. Modular systems (UASB and ARC) sewage treatment showed a good performance, with high values of removing SSed (98.72; 98.36), and COD (72.37; 75.79) with no significant difference between the treatment systems. It was noted also that the flower beds with wastewater reuse treatments promoted change in the fertility of the soil, with the increase in base saturation values (V%) at the depth of 0-20 cm. Analyzing the effects on culture in three plantations, there was a better development in the flower beds with the effluent treatments, when compared with the other treatments mainly in the second and third planting. Modular wastewater treatment system associated with the agricultural reuse of wastewater can add value to their agricultural production, and can be used throughout the year even in places where there is scarcity of rainwater / Doutorado / Agua e Solo / Doutor em Engenharia Agrícola

Waterborne diarrhoeal infections continue to be a major health setback in developing countries, especially in rural areas which lack adequate supply of portable water and sanitation facilities. Globally, waterborne diarrhoeal infections occur with an estimated mortality rate of 10–25 million deaths per year, 95% of which are children under the age of 5 years. The Vibrio species is one of the major groups of enteric pathogens that are responsible for diarrhoeal infections. Many strains of these bacterial species continue to cause epidemics of diarrhoea throughout the world. In this study, the prevalence of Vibrio pathogens in wastewater final effluents was assessed. Wastewater final effluent and discharge point samples were collected monthly between September 2012 and August 2013. All samples were collected aseptically using sterile 1 L Nalgene bottles containing 0.5 ml of sterile sodium thiosulphate solution and transported on ice to the laboratory for analyses within 6 h of collection. The membrane filtration method was used for enumeration of presumptive Vibrio densities on thiosulfate citrate bile salt (TCBS) agar plates. Polymerase chain reaction (PCR) was then used to confirm the identities of the presumptive Vibrio species using the species-specific primers. The confirmed isolates were further subjected to molecular characterization to confirm their respective pathotypes. Presumptive Vibrio densities varied from 0 to 2.11 × 102 cfu/100 ml. Out of 300 confirmed Vibrio isolates; 13.3% (40/300) were Vibrio fluvialis, 22% (66/300) were confirmed to be Vibrio parahaemolyticus, and 24.7% (74/300) proved to be Vibrio vulnificus, and 40% (120/300) were other Vibrio species which were not assessed for in this study. The strains of Vibrio fluvialis were found to exhibit 100% resistance to Polymixin and Tetracycline. However, Gentamicin was active against all the three Vibrio species selected for the purpose of this research. The recovery of Vibrio species in the discharged effluents throughout the sampling period even in adequately disinfected effluents is not acceptable considering the fact that Vibrio is a pathogenic bacterium. The findings of this study underline the need for constant monitoring of the microbiological qualities of discharged effluents and might also be suggestive for a review of the disinfection methods used at the treatment works.

Integrated Algae Pond Systems (IAPS) are a derivation of the Oswald designed Algal Integrated Wastewater Pond Systems (AIWPS®) and combine the use of anaerobic and aerobic bioprocesses to effect wastewater treatment. IAPS technology was introduced to South Africa in 1996 and a pilot plant designed and commissioned at the Belmont Valley WWTW in Grahamstown. The system has been in continual use since implementation and affords a secondarily treated water for reclamation according to its design specifications which most closely resemble those of the AIWPS® Advanced Secondary Process developed by Oswald. As a consequence, and as might be expected, while the technology performed well and delivered a final effluent superior to most pond systems deployed in South Africa it was unable to meet The Department of Water Affairs General Standard for nutrient removal and effluent discharge. The work described in this thesis involved the design, construction, and evaluation of several tertiary treatment units (TTU') for incorporation into the IAPS process design. Included were; Maturation Ponds (MP), Slow Sand Filter (SSF) and Rock Filters (RF). Three MP's were constructed in series with a 12 day retention time and operated in parallel with a two-layered SSF and a three-stage RF. Water quality of the effluent emerging from each of these TTU's was monitored over a 10 month period. Significant decreases in the chemical oxygen demand (COD), ammonium-N, phosphate-P, nitrate-N, faecal coliforms (FC) and total coliforms (TC) were achieved by these TTU's. On average, throughout the testing period, water quality was within the statutory limit for discharge to a water course that is not a listed water course, with the exception of the total suspended solids (TSS). The RF was determined as the most suitable TTU for commercial use due to production of a better quality water, smaller footprint, lower construction costs and less maintenance required. From the results of this investigation it is concluded that commercial deployment of IAPS for the treatment of municipal sewage requires the inclusion of a suitable TTU. Furthermore, and based on the findings presented, RF appears most appropriate to ensure that quality of the final effluent meets the standard for discharge.

Sewage effluents disposed into the marine environment from De Beers Marine Namibia diamond mining vessels have the potential to cause endocrine disruptive effects in marine organisms. Endocrine disruption refers to the alteration of the normal functioning of the endocrine system and various chemicals have the ability to mimic hormones, effecting endogenous hormone synthesis, transport, receptor interaction and intracellular signaling. The potential endocrine disruptive effects, caused by the release of different types of sewage effluents into the ocean, on fish species is a concern due to the commercial importance of fish species found in the mining area e.g. hake, sole, horse mackerel. Increased awareness of marine environmental degradation due to the presence of chemical contaminants has resulted in research being done on early warning systems, in the form of biomarkers. Cytochrome P450 monooxygenase 1A (CYP1A) and vitellogenin (vtg) are important proteins found in fish liver and blood, that have been used as biomarkers for the detection of pollutants in fish. CYP1A is a subfamily of the P450 superfamily of enzymes and catalyzes the oxidation, hydrolysis and reduction of exogenous and endogenous compounds (phase I reactions) and thus has the capacity to regulate the metabolism of several organic contaminants. CYP1A expression is altered by exposure to planar xenobiotic compounds e.g. polyaromatic hydrocarbons. Vtg is an important precursor for egg yolk proteins and plays a role in the growth and development of an oocyte. Expression of this protein is altered upon exposure to estrogenic compounds. The aim of this project was to isolate CYP1A from fish liver by differential centrifugation and optimize conditions for the CYP1A-mediated ethoxyresorufin-Odeethylase (EROD) assay and western blot analysis (to assess CYP1A expression). Another aim of this study was to evaluate the potential effects of biologically disruptive chemicals from sewage effluents, discharged into the marine environment, on the expression of CYP1A in two species of hake, Merluccius capensis and M. paradoxus (Cape hake). CYP1A in Cape hake is approximately a 60 kDa protein and the highest EROD activity was detected in the microsomal fraction after differential centrifugation. Optimal EROD assay conditions were observed at pH 7.5, a temperature of 25 °C, 10 μl of sample and a reaction time of 30 seconds. Enzyme stability assays indicated a drastic decrease in enzyme activity after 30 seconds. The EROD assay was not NADPH dependent but was limited by NADPH supply, with an increase of 300% in EROD activity being observed with the addition of 0.1 M exogenous NADPH. The addition of dicumarol (40 μM), a phase II enzyme inhibitor, showed a 232% increase in EROD activity. This is because dicumarol inhibited enzymes with the capacity to metabolize the product (resorufin) of the EROD reaction. With regard to western blot analysis, the optimal primary (rabbit antifish CYP1A peptide) and secondary (anti-mouse/rabbit antibody-horseradish peroxidase conjugate (POD)) antibody dilutions were determined to be 1:1000 and 1:5000, respectively. The comparison of CYP1A expression in Cape hake samples from De Beers Marine mining area and reference sites showed higher EROD activity (16.29 ± 0.91 pmol/min) in fish samples from the mining area in comparison to the reference site (10.42 ± 2.65 pmol/min). Western blot analysis was in agreement with the EROD assay results and a higher CYP1A expression was observed in fish from the mining sites. The increased CYP1A expression observed in fish from the mining area is not definitively an indication of a pollutant effect in the environment, as several environmental and biological factors (e.g. photoperiod and age) must also be considered before reaching this conclusion. Another aim of this study was to purify vtg from Cape hake blood samples. Cape hake vtg was purified from fish plasma by selective precipitation with MgCl2 and EDTA. Precipitated sample was subjected to anion exchange chromatography using fast protein liquid chromatography (FPLC). Vtg eluted as two broad peaks and had a molecular weight above 200 kDa. SDS-PAGE analysis also resolved smaller molecular weight proteins below 70 kDa, which were thought to be vitellogenin cleavage proteins, lipovitellin and phosphovitins. Western blot analysis was performed; however, it did not produce any conclusive results. The purification of vtg enables further studies in characterizing this protein and developing assay aimed at detecting estrogenic pollutants in the marine environment