Thesis submitted in compliance with the requirements for the Doctoral Degree in Technology: Biotechnology at the Durban Institute of Technology, 2003. / Globally, wastewaters emanating from edible oil manufacturers contain high organic (BOD & COD) and phosphate loads and known for creating shock-loading problems for the receiving wastewater treatment installations. Discharge of poor quality final effluents also negatively impact on and cause eutrophication of natural water sources such as rivers and dams. In South Africa, a large concentration ofthe edible industries are localized in the Pietermaritzburg region of Kwa-Zulu Natal and have been regularly associated with discharge of poor quality final effluent that did not subscribe to municipal regulation standards. Current treatment of choice for wastewater's in the edible oil industry have been limited primarily to dissolved air flotation combined with the use of chemical coagulants or physical separation of oil and grease via a gravity fat trap and subsequent pH correction. These physico-chemical methods have achieved limited success and the emulsified grease tends to clog sewer pipes and pumps producing poor quality effluents. Therefore, the aim of the current research was to develop suitable treatment technology focussing on adapting activated sludge process to remediate edible oil effluents and determine the microbial community of the process using novel molecular techniques. / D

The microbial populations in the Hoechst-Celanese activated sludge were examined. Heterotrophs, denitrifiers, sulfate-reducers, protozoa and filamentous bacteria were enumerated. Variations in microbial populations were compared with influent and effluent constituent concentrations, and with aeration basin characteristics, such as dissolved oxygen and F/M ratio, to determine whether any microbial type could be used by plant operators to monitor process performance. Results indicated that filamentous bacteria may be useful to plant operators for monitoring process performance because an inverse relationship between filamentous bacteria, food-to-microorganism ratio and sludge volume index was suggested by this study. Protozoa may also be useful for operators, although more data is needed. Microthrix parvicella and Type 0041 were the most common filament types. Filament Type 1701 was most prevalent during a period of low dissolved oxygen. A strong relationship between stalked ciliates and effluent quality was mention in the literature, but was not found in this study. Enumeration methods were evaluated. / Master of Science

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnolgy, Durban University of Technology, 2007 xviii, 143 leaves / Activated sludge flocs are responsible for flocculation, settling and dewaterability. It is important to maintain the growth off loc-forming bacteria for efficient sludge settleability and compaction for good quality effluent. Filamentous bacteria on the other hand are believed to provide rigid support network or backbone upon which floc-forming bacteria adhere to form stable activated sludge flocs (Wilderer et al., 2002; Ramothokang et al., 2003). Filamentous bacteria can also be detrimental to the process when they outgrow floc-forming bacteria. Morphologically filamentous bacteria are at an advantage as they have higher outward growth velocity and can extend freely to bulk liquid substrate. Proliferation of filamentous bacteria causes foaming and bulking (Martins et al., 2004). Although chemical alleviation measures to circumvent bulking are present, they are symptomatic (Chang et al., 2004). Eikelboom (1975) developed the first identification keys for the classification of filamentous bacteria that is primarily based on morphological characteristics and microscopic examination. Although very useful, this type of identification has its limitations. For instance some filamentous bacteria can change morphology in response to changes in the environment and although some of them can be morphologically similar they may vary considerably in their physiology and taxonomy (Martins et al., 2004). A vast number of filamentous bacteria are still very poorly understood which could be due to the problems of cultivation due to their slow growing nature and maintenance of cultures (Rossetti et al., 2006). This limitation necessitates a molecular approach to resolve the taxonomy of filamentous bacteria as it is a culture-independent technique which is highly accurate. This project was undertaken to verify the identity of pure cultures of filamentous bacteria isolated previously through the application of molecular techniques. The 16S rDNA are conserved regions in bacterial cells and they can be extracted and specific nucleic acid fragments amplified. Denaturation gradient gel electrophoresis enabled the separation of fragments of identical length but different size and served as an indication of purity (Muyzer et al., 1993).

Thesis (MSc)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: Large volumes of wastewater are generated annually during the grape harvest season from various processing and cleaning operations at wineries, distilleries and other wine-related industries. South African regulatory bodies dictate that wastewater should have a pH of 5.5 to 7.5 and a chemical oxygen demand (COD) lower than 75 mg/L. However, winery wastewater has a typical pH of 4 to 5 and a COD varying between 2 000 and 12 000 mg/L. Urban wineries channel the wastewater to local sewage treatment facilities and are often heavily fined for exceeding governmental requirements. Rural wineries usually have little or no treatment operations for their wastewater and it is often irrigated onto crops, which may result in environmental pollution and contamination of underground water resources. Various criteria are important in choosing a wastewater treatment system, such as an ecofriendly process that is flexible to withstand various concentration loads and characteristics, requiring low capital and operating costs, minimal personal attention and do not require too much land. In this study, a large variation in COD, pH and chemical composition of the winery wastewater was observed that could be related to varying factors such as the harvest load, operational procedures and grape variety. Wastewater from destemming and pressing operations contained higher concentrations of glucose, fructose and malic acid, which originated from the grape berries. The fermentable sugars (glucose and fructose) contributed to almost half of the COD with a smaller contribution from ethanol and acetic acid. The low pH can be ascribed to relative high concentrations of organic acids in the wastewater. The efficacy of biological treatment systems depends strongly on the ability of microorganisms to form biofilm communities that are able to degrade the organic compounds in the wastewater. Preliminary identification of microorganisms that naturally occur in winery wastewater indicated the presence of various bacterial and yeast species that could be effective in the biological treatment of the wastewater. When evaluated as pure cultures under aerobic conditions, some of the yeast isolates effectively reduced the COD of a synthetic wastewater, whereas the bacterial isolates were ineffective. The most effective yeast isolates were identified as Pichia rhodanensis, Kloeckera apiculata, Candida krusei and Saccharomyces cerevisiae. Our search for cost-effective biological treatment systems led to the evaluation of a Rotating Biological Contactor (RBC) for the treatment of winery wastewater. The RBC was evaluated on a laboratory scale with 10% (v/v) diluted grape juice and inoculated with a mixed microbial community isolated from winery wastewater. The results showed a reduction in the COD that improved with an extended retention time. Evaluation of the RBC on-site at a local winery during the harvest season resulted on average in a 41% decrease in COD and an increase of 0,75 pH units. RFLP analysis of the biofilm communities within the RBC confirmed a population shift in both the bacterial and fungal species during the evaluation period. The most dominant yeast isolates were identified with 18S rDNA sequencing as Saccharomyces cerevisiae, Candida intermedia, Hanseniaspora uvarum and Pichia membranifaciens. All these species are naturally associated with grapes and/or water and with the exception of Hanseniaspora uvarum, they are able to form either simple or elaborate pseudohyphae. / AFRIKAANSE OPSOMMING: Groot hoeveelhede afloopwater word jaarliks gedurende die druiwe-oestyd deur verskeie prosessering- en skoonmaakoperasies deur wynkelders, distilleer- en ander wynverwante industrieë gegenereer. Suid-Afrikaanse beheerliggame vereis dat afloopwater ‘n pH van 5.5 tot 7.5 en ‘n chemiese suurstofbehoefte (COD) van minder as 75 mg/l moet hê. Kelderafloopwater het egter gewoonlik ‘n pH van 4 tot 5 en ‘n COD van 2 000 tot 12 000 mg/L. Stedelike wynkelders voer die afloopwater na ń plaaslike rioolsuiweringsaanleg wat dikwels tot swaar boetes vir oortreding van die wetlike vereistes lei. Plattelandse wynkelders het gewoonlik min of geen behandelingsprosesse vir hul afloopwater nie en gebruik die water dikwels vir gewasbesproeiing, wat tot omgewingsbesoedeling en kontaminasie van ondergrondse waterbronne kan lei. Verskeie kriteria is belangrik in die keuse van ‘n waterbehandelingstelsel, byvoorbeeld ‘n omgewingsvriendelike proses wat verskillende konsentrasieladings en samestellings kan hanteer, ‘n lae kapitaal- en bedryfskoste en minimale persoonlike aandag vereis en min ruimte benodig. Hierdie studie het getoon dat kelderafloopwater ‘n groot variasie in COD, pH en chemiese samestelling het wat met wisselende faktore soos die oeslading, operasionele prosesse en selfs die druifkultivar verband kan hou. Afloopwater van ontstingeling- en parsoperasies het hoër konsentrasies glukose, fruktose en appelsuur wat van die druiwekorrels afkomstig is. Die fermenteerbare suikers (glukose en fruktose) dra tot amper 50% van die COD by, met ‘n kleiner bydrae deur etanol en asynsuur. Die lae pH kan grootliks aan organiese sure in die afloopwater toegeskryf word. Die effektiwiteit van biologiese behandelingstelsels steun sterk op die vermoë van mikroorganismes om biofilmgemeenskappe te vorm wat die organiese verbindings in die afloopwater kan afbreek. Voorlopige identifikasie van mikro-organismes wat natuurlik in wynafloopwater voorkom, het die teenwoordigheid van verskeie bakteriese en gisspesies aangedui. Evaluering van hierdie isolate onder aërobiese toestande het getoon dat sommige van die gis-isolate die COD van ‘n sintetiese afloopwater effektief kon verlaag, terwyl die bakteriese isolate oneffektief was. Die mees effektiewe gis-isolate is as Pichia rhodanensis, Kloeckera apiculata, Candida krusei en Saccharomyces cerevisiae geïdentifiseer. Ons soektog na ‘n koste-effektiewe biologiese behandelingsisteem het tot die evaluering van ‘n ‘Rotating Biological Contactor’ (RBC) vir die behandeling van afloopwater gelei. Die RBC is op laboratoriumskaal met 10% (v/v) verdunde druiwesap geëvalueer en met ‘n gemengde mikrobiese gemeenskap wat uit afloopwater geïsoleer is, innokuleer. Die resultate het ‘n verlaging in die COD getoon wat met ‘n langer retensietyd verbeter het. Evaluering van die RBC by ‘n plaaslike wynkelder gedurende die oesseisoen het gemiddeld ‘n verlaging van 41% in die COD en ‘n verhoging van 0,75 pH eenhede getoon. RPLP analise van die biofilmgemeenskappe in die RBC het ‘n bevolkingsverskuiwing in beide die bakteriese en swamspesies aangetoon. Die mees dominante gisspesies is met 18S rDNA volgordebepaling as Saccharomyces cerevisiae, Candida intermedia, Hanseniaspora uvarum en Pichia membranifaciens geïdentifiseer. Al hierdie spesies word gewoonlik met druiwe en/of water geassosieer en is, met die uitsondering van Hanseniaspora uvarum, in staat om òf eenvoudige òf komplekse pseudohife te vorm.

Coliform bacteria have long been used to indicate fecal contamination of food, water, and solid surfaces, and thus the presence of a health hazard. In this study, the in-mill water and external effluent treatment systems of seven typical Canadian pulp and paper mills were shown to support the growth of numerous coliforms, especially Klebsiella spp, Escherichia coli, Enterobacter spp., and Citrobacter spp . Mill coliforms were shown to be not just simple transients from feedwater or furnish (wood), but to be continuously growing, especially in the primary clarifiers. Therefore, coliforms and fecal coliforms cannot be used as fecal contamination indicators in pulp and paper mill water and effluent treatment systems. / N2-fixing coliform populations were detected in mill water systems and were analyzed using N2-fixation assays and nitrogenase gene (nifH) probing. Both active in situ populations and cultured microbial isolates were tested. Active N2-fixation was demonstrated in six primary clarifiers. Measurement of the numbers and composition of the total culturable bacterial community in a primary clarifier revealed that approximately 50% of all aerobic cells contained nifH , of which >90% were Klebsiella. Coliforms growing on MacConkey agar plates from the primary clarifier were all identified as Klebsiella and 100% of these Klebsiella contained the nifH gene. Preliminary estimates indicate that the amount of N2 fixed per day is substantial in some clarifiers.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnolgy, Durban University of Technology, 2007 xviii, 143 leaves / Activated sludge flocs are responsible for flocculation, settling and dewaterability. It is important to maintain the growth off loc-forming bacteria for efficient sludge settleability and compaction for good quality effluent. Filamentous bacteria on the other hand are believed to provide rigid support network or backbone upon which floc-forming bacteria adhere to form stable activated sludge flocs (Wilderer et al., 2002; Ramothokang et al., 2003). Filamentous bacteria can also be detrimental to the process when they outgrow floc-forming bacteria. Morphologically filamentous bacteria are at an advantage as they have higher outward growth velocity and can extend freely to bulk liquid substrate. Proliferation of filamentous bacteria causes foaming and bulking (Martins et al., 2004). Although chemical alleviation measures to circumvent bulking are present, they are symptomatic (Chang et al., 2004). Eikelboom (1975) developed the first identification keys for the classification of filamentous bacteria that is primarily based on morphological characteristics and microscopic examination. Although very useful, this type of identification has its limitations. For instance some filamentous bacteria can change morphology in response to changes in the environment and although some of them can be morphologically similar they may vary considerably in their physiology and taxonomy (Martins et al., 2004). A vast number of filamentous bacteria are still very poorly understood which could be due to the problems of cultivation due to their slow growing nature and maintenance of cultures (Rossetti et al., 2006). This limitation necessitates a molecular approach to resolve the taxonomy of filamentous bacteria as it is a culture-independent technique which is highly accurate. This project was undertaken to verify the identity of pure cultures of filamentous bacteria isolated previously through the application of molecular techniques. The 16S rDNA are conserved regions in bacterial cells and they can be extracted and specific nucleic acid fragments amplified. Denaturation gradient gel electrophoresis enabled the separation of fragments of identical length but different size and served as an indication of purity (Muyzer et al., 1993).

No description available.

Le bassin versant de l’Escaut (20 000 km²) est caractérisé par forte densité de population (plus de 500 habitants par km2) et une activité agro-pastorale intensive. Les rivières de ce bassin sont sévèrement affectées par les rejets d’eaux usées domestiques, les effluents d’élevage et les eaux de ruissellement des sols agricoles. Le but de cette étude est :(i) d’évaluer la qualité microbiologique de ces rivières ;(ii) d'identifier et de quantifier les différentes sources de contamination fécale à l’échelle du bassin de l’Escaut; (iii) d’étudier les processus qui contrôlent le devenir des bactéries fécales en rivière; (iv) de développer des modèles numériques sur la base des travaux expérimentaux permettant de prédire la concentration des bactéries indicatrices dans les rivières du bassin de l’Escaut.<p>L’évaluation de la qualité microbiologique des principales rivières du bassin est basée sur le dénombrement de deux indicateurs de contamination fécale (Escherichia coli et entérocoques intestinaux). Les abondances des deux indicateurs dans les principales rivières du bassin indiquent très clairement que les eaux et les sédiments de ces rivières sont fortement contaminés par des micro-organismes entériques. Les sources prédominantes de la pollution fécale de ces rivières sont les rejets des effluents des stations d’épuration. Les niveaux de contamination les plus élevés sont observés dans la Senne en aval de Bruxelles et s’expliquent par le faible débit de la Senne comparé aux débits des effluents des deux stations d’épuration de Bruxelles. Les niveaux de contamination atteignent leur maxima à l’aval de Bruxelles par temps de pluie en raison des surverses de réseaux unitaires.<p>Les connaissances acquises sur les apports des bactéries indicatrices par les sources ponctuelles et les sources diffuses et sur le devenir des bactéries indicatrices ont permis de développer un module décrivant la dynamique des E. coli dans les rivières. Ce module est original par le fait de considérer trois compartiments de bactéries fécales (libres, attachées aux particules dans la colonne d’eau et présentes dans les sédiments) qui sont affectés différemment par les processus de transport et de disparition. Ce module a été couplé à deux modèles décrivant l’hydrodynamique respectivement de l’ensemble des rivières du bassin (SENEQUE-EC) et de la partie fluviale de l’Escaut sous l’influence de la marée et son estuaire (SLIM-EC2). Ces deux modèles permettent de décrire la distribution temporelle et spatiale des E. coli dans les eaux de surfaces et de prévoir les modifications de la qualité microbiologique des eaux suite à des changements de gestion des eaux usées. <p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished