Thesis (MSc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: The "classical" propionibacteria are used in a variety of natural dairy fermentations where they produce natural preservatives (propionic and acetic acids and bacteriocins) and large amounts of vitamin B12. The extracellular polysaccharide (ECP) producing ability of these bacteria also make them of special interest to the food and waste water management industries as the ECP has been illustrated to playa role in the initial granule formation in upflow anaerobic bioreactor systems. There is little known on the ECP production by propionibacteria and in this study different environmental conditions that influence ECP production were studied. Nineteen different Propionibacterium strains were examined in terms of ECP production and Propionibacterium strain 278 was identified as the best ECP producer. Further studies were only done on this strain because of its high ECP production and because it was originally isolated from an anaerobic digester. The influence of temperature, pH and sucrose concentration was determined through the measurement of ECP production and medium viscosity. It was found that more ECP was produced at temperatures lower than the optimum for growth with the optimum being between 22° and 25°C. Lower initial pH conditions of the growth medium (below pH 7.0) were found to inhibit ECP production and the influence when the initial pH values were between 7.0 and 8.5, was not significant. A higher carbon: nitrogen ratio, when 8% sucrose was added, was also found to enhance the ECP production. The upflow anaerobic sludge bed (UASB) bioreactor process depends on the upward movement of soluble matter through a blanket of active methanogenic granular sludge. The long start-up times as a result of the slow granulation process, as well as the need for a speedy replacement of granules once they have been washed out of the system, are limitations that restrict the general application of this excellent waste water treatment technology. Full exploitation of this biomass immobilisation technique can thus not be realised until the granule formation conditions are defined and optimised. The precise nature of the mechanisms involved in the formation of granules and the reason for their stability, is still not fully understood. It was hypothised by Britz et al. in 1999 that, through the implementation of environmental 'stress' conditions, a shift in the population dynamics of the anaerobic community can be obtained. This results in a concurrent increase in ECP formation that appears to enhance aggregate formation. In the second study it was found that, when 'stress' conditions were applied to already formed granules, the Gram-positive lactate-utilising acidogenic population gained an advantage and more propionic acid producing bacteria were present. The propionic and acetic acid concentrations were also found to increase, and concurrently, a decrease in the growth medium pH occurred. This confirms part of the granulation hypothesis that, when granules are 'stressed', the acidogenic population dynamics change and the lactate-utilising population responds to the gradual decrease in pH and the more acid-tolerant propionic acid producing bacteria gain a competitive advantage resulting in the increase in the propionic acid concentration. When propionibacteria were added to raw sludge during the granule production process, the granules were found to be more active than when nopropionibacteria had been added. This was probably due to the ECP formation by the propionibacteria that enhances the aggregation of the granules. Enhanced granulation was thus found in the batch systems with the fatty acids formed in correlation with the model for granulation. A good correlation was evident between the hypothesis and the experimental data and the hypothesis was partially verified in this study. / AFRIKAANSE OPSOMMING: Die "klassieke" propionibakterieë word in 'n verskeidenheid van natuurlike suiwel fermantasies gebruik waarin hulle verantwoordelik in vir die produksie van natuurlike voedsel preserveermiddels (propioonsuur, asynsuur en bakteriosiene) en groot hoeveelhede vitamiene B12. Die Ekstra Sellulêre Pollisakkaried (ESP) produserende eienskap van hierdie groep bakterieë maak hulle ook van belang in die voedsel en afvoerwater beheer industrieë, aangesien gevind is dat ESP 'n rol speel in die aanvanklike granule formasie in anaerobiese bioreaktor sisteme. Daar is nog baie min bekend oor die ESP produksie van propionibakterieë en in hierdie studie is verskeie omgewings faktore wat die ESP produksie beïnvloed, bestudeer. Negentien verskillende Propionibakterium stamme was bestudeer in terme van ESP produksie en Propionibakterium stam 278 was geïdentifiseer as die stam wat die meeste ESP produseer. Verdere studies was op hierdie stam gedoen na aanleiding van sy hoë ESP produksie en omdat dit oorspronklik uit 'n anaerobiese verteerder geisoleer is. Die invloed van termperatuur, pH en sukrose konsentrasie was bepaal deur die meting van die ESP produksie en die medium viskositeit. Dit was gevind dat meer ESP geproduseer was by temperature laer as die optimum vir groei, met die optimum temperatuur tussen 22° en 25°C. Dit is ook gevind dat laer aanvangs groei-medium pH (laer as pH 7.0), ESP produksie inhibeer. Die invloed van die aanvangs groei-medium pH tussen 7.0 en 8.5 was egter nie betekenisvol nie. Dit is ook gevind dat 'n hoër koolstof tot stikstof verhouding, verkry deur die byvoeging van 8% sukrose, die ESP produksie verhoog. Die "upflow anaerobic sludge blanket" (UASB) proses vind plaas as gevolg van die opwaarste beweging van opgeloste organiese materiaal deur 'n granule bed van aktiewe metanogeniese granulêre slyk. Die lang 'start-up' tye as gevolg van die stadige granulasie proses, en die nodigheid om 'n vinnige verplasing van granules te hê nadat dit uit die sisteem gewas is, is beperkings wat die algemene toepassing van hierdie fantastiese afvoerwater tegnologie, strem. Volle implementering van hierdie biomassa immobilisereings tegniek kan dus nie plaasvind voordat die granule formasie gedefinieer en geoptimiseer is nie. Die presiese eienskappe van die meganismes betrokke en die formasie van die granules en die rede vir hul stabiliteit word egter nog nie ten volle verstaan nie. Volgens 'n hipotese deur Britz et al. (1999), vind 'n verskuiwing in die populasie dinamika van die anaerobiese gemeenskap plaas tydens die implementasie van omgewings 'stress' toestande. Die resultaat is 'n verhoging in ESP produksie en 'n gevolglike verbetering in die granulasie proses. In die tweede studie was dit gevind dat, wanneer 'stress' toestande op die reeds gevormde granulasie toegepas word, die Gram-positiewe laktaat-benuttende asetogeniese populasie voordeel geniet en meer propioonsuur produserende bakterieë was teenwoordig. Die propioonsuur en asynsuur konsentrasies het ook verhoog en met 'n gevolglike daling in die groei-medium se pH. Dit bevestig 'n gedeelte van die hipotese dat, wanneer die granules onder 'stress' geplaas word, die asetogeniese populasie dinamika verander en die laktaat-benuttende populasie reageer tot die gedeeltelike afname in pH. Die meer suur-tolerante propioonsuur produserende bakterieë verkry 'n kompeterende voordeel en gevolglik is daar 'n verhoging in propioonsuur konsentrasie. Propionibakterieë was gevoeg by die onbehandelde slyk gedurende die granule produksie proses, en daar is gevind dat meer aktiewe granules gevorm word as andersins. Dit is moontlik as gevolg van die die ESP produksie van propionibakterieë wat die granulasie versnel het. Verbeterde granulasie was dus verkry in die sisteme waar propionibakterieë bygevoeg is. Vetsuur analises het gedui dat die gevormde vetsure ook in korrelasie was met die model van granulasie. Goeie korrelasie was dus verkry tussen die hipotese en die eksperimentele data en die hipotese is gedeeltelik bewys in hierdie studie.

Thesis (MSc Food Sc )--Stellenbosch University, 2000. / ENGLISH ABSTRACT: The potential use of the upflow anaerobic sludge blanket (UASB) bioreactor is limited by the extended start-up periods due to the time-consuming granulation process and the fact that seeding inoculum is not freely available in most developing countries like South Africa. The mass cultivation of granules would provide the waste water treatment industry with suitable seeding inoculum and, therefore, an efficient waste water treatment option would be more easily available. By applying 'stress' conditions on a bioreactor system seeded with raw anaerobic sludge, the population dynamics of the anaerobic community change and the acidogens start to produce extracellular polymers, which in turn enhance the granulation process by providing a matrix for the bacterial cells to adhere to. These "environmental stress" conditions include changes in the C:P:N ratio's. The aim of this study was, therefore, to assess the impact of different carbon and nitrogen sources on the enhancement of granulation in a batch system, and to determine the best preservation technique in terms of retainment of activity. It was found that the carbon source and concentration had a significant influence on batch granule enhancement. Low concentrations (2.g.l ̄ ¹) of glucose gave the best granule enhancement over a 14 day incubation period. Fruit cocktail effluent was found to be a cheap and effective carbon source for batch granule cultivation. It was found that different nitrogen sources did not have the same impact on granule enhancement, however, urea, at all concentrations tested, gave the best granule enhancement. A major problem encountered during the study was the standardisation of the inoculum. Significant granule enhancement comparisons between the different carbon sources were impeded by the lack of a suitable and reliable form of sludge standardisation. Future research needs to address this problem of the standardisation of the sludge inoculum as this would lead to more kinetically comparable results. Mass granule culturing will require granule preservation without risking the loss of activity. In the second study, six different preservation techniques and storage periods were evaluated in terms of the retainment of activity. These involved freeze-drying, vacuum freeze-drying, vacuum-drying, freezing, cold storage and room temperature preservation. Activity testing was used for comparing the efficiency of the different preservation techniques in terms of the tempo of biogas and methane production. Freeze-drying the granules, with storage for up to 90 days was found to give the best retainment of activity, followed by vacuum freeze-drying. The room temperature preserved samples showed a sudden increase in activity by day 120, which could possibly be explained by cell hydrolysis of the granules after day 90. The highest activity was achieved after 10 h of incubation, and it was, therefore, suggested that activity testing for evaluation purposes should use an incubation time of only 10 h. Furthermore, only the basic test medium with added glucose should be used for activity testing as it was observed that the addition of lactate and acetic acid played no decisive role in determining the level of activity of the granules. This study recommends the use of low concentrations of glucose for optimum granule enhancement during the mass cultivation of granules, and the addition of low concentrations of lactate to ensure a stable system with no acidification. It is also advisable to use a standardised sludge inoculum, as this will allow more efficient comparisons. Freeze-drying is recommended as preservationtechnique as this technique showed the best retainment of activity. A storage period of 90 days is, however, too short to be of much use for the industry. This will have to be investigated, together with the phenomenon of increased activity after 90 days as shown by the room temperature preserved granules. / AFRIKAANSE OPSOMMING: Die potensiële gebruik van die "uptflow anaerobic sludge blanket" (UASB) bioreaktor word beperk deur verlengde aansitprosedure as gevolg van die tydsame granulasieproses en die feit dat geskikte inokulums nie vrylik beskikbaar is in die meeste ontwikkelende lande, bv. Suid-Afrika nie. Die massa-kweking van granules sal die afvalwater-industrie voorsien van 'n geskikte inokulum, en sodoende 'n effektiewe opsie vir die behandeling van afvalwater beskikbaar stel. Deur die aanwending van "stres" toestande op 'n reaktor, wat geinokuleer is met rou anaerobe slyk, vind daar 'n verandering in die samestelling van die anaerobe populasie plaas. Die laktaat-benuttende asidogene begin om ekstrasellulêre polisakkariede te produseer wat die granulasieproses versnel deur 'n matriks te voorsien waaraan die bakteriële selle kan heg. Hierdie "omgewings stres" toestande sluit veranderinge in die C:P:N verhoudings in. Die doel van hierdie studie was om die invloed van verskillende koolstof- en stikstofbronne op die vermeerdering van granules te bepaal, asook om die beste preserveringstegniek in terme van die behoud van aktiwiteit te bepaal. Daar is gevind dat die koolstofbron en konsentrasie 'n betekenisvolle invloed op granule-vermeerdering het. Lae konsentrasies glukose (2 g.I ̄ ¹) het tot die grootste vermeerdering in granules oor 'n 14 dae inkubasieperiode gelei. Vrugtekelkie-afvalwater is geidentifiseer as 'n goedkoop en effektiewe koolstofbron wat ook gebruik kan word vir die kweking van granules. Voorts is gevind dat die stikstofbron en konsentrasie nie so 'n groot invloed op granule-vermeerdering het nie. Urea het nietemin die beste granule-vermeerdering vir al die konsentrasies wat getoets is, gegee. Standaardisasie van die inokulum was 'n groot probleem gedurende die studie. Betekenisvolle vergelykings tussen die granule-vermeerderings verkry met die verskeie groeimedia is bemoeilik deur die afwesigheid van 'n geskikte en betroubare standaardisasie-metode. Toekomstige navorsing moet hierdie probleem aanspreek aangesien dit sal lei tot meer kineties vergelykbare resultate. Massa-kweking van granule sal 'n metode van granule-preservering vereis sonder enige verlies van aktiwiteit. In die tweede studie is ses verskillende preserverings-tegnieke en opbergings-periodes in terme van die behoud van aktiwiteit geëvalueer. Die tegnieke sluit in: vriesdroging; vakuum-vriesdroging; vakuumdroging; bevriesing; koelopberging en kamertemperatuur-preservering. Aktiwiteitstoetsing is gebruik vir die vergelyking van die effektiwiteit tussen die verskillende preserverings-tegnieke in terme van die tempo van biogas- en metaanproduksie. Die granules wat met behulp van die vriesdroogtegniek gepreserveer is, het die beste behoud van aktiwiteit getoon, gevolg deur die vakuum-gevriesdroogde granules. Die monsters wat by kamertemperatuur gepreserveer is, het 'n skielike toename in aktiwiteit na 120 dae van opberging getoon en 'n moontlike verklaring vir hierdie verskynsel kan sel-hidrolise van die granules na 90 dae van opberging wees. Die meeste aktiwiteit is behaal na 'n 10 h inkubasietyd, en dus word hierdie inkubasietyd aanbeveel indien aktiwiteitstoetse vir evaluerings-doeleindes gedoen word. Verder word aanbeveel dat slegs die glukose-verrykte basiese toetsmedia vir aktiwiteitstoetsing gebruik word aangesien die byvoeging van laktaat en asynsuur geen noemenswaardige rol speel in die bepaling van die vlak van aktiwiteit van die granules nie. Hierdie studie beveel die gebruik van lae glukosekonsentrasies aan vir optimale vermeerdering van granules tydens die massakweking daarvan, asook die byvoeging van lae konsentrasies laktaat om 'n stabiele sisteem met geen versuring te verseker. Die gebruik van 'n gestandaardiseerde slyk-inokulum word sterk aanbeveel aangesien dit meer vergelykbare resultate lewer. Vriesdroging (as preserveringstegniek) het die beste behoud van aktiwiteit in die granules getoon en word dus as preserveringstegniek aanbeveel. 'n Opbergingsperiode van 90 dae is egter te kort om van veel waarde vir die industrie te wees. Hierdie probleem, asook die verskynsel van 'n verhoging in aktiwiteit na 90 dae van die granules wat by kamertemperatuur gepreserveer is, moet verder ondersoek word.

Thesis (MSc Food Sc)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: The start-up period of Upflow Anaerobic Sludge Bed (UASB) reactors can significantly be reduced by enhancing the time-consuming granulation process through the batch cultivation of anaerobic granular sludge and thus seeding the reactor with this cultivated granular sludge, instead of raw anaerobic sludge. The precise mechanism for granule formation is not well known, but it is believed that extracellular polymers (ECP) play a critical role in the granulation process. Information on the precise role of ECP is also limited and no universal standardised method for ECP extraction is used at present. Therefore, comparison of results from different researchers has to be made with great caution. The objectives of this study were to evaluate an ECP extraction methód so as to optimise the extraction time, and then to correlate ECP composition of UASB granules with granule metabolic activity. The impact of changes in the environmental conditions, such as sludge sources (Paarl and Kraaifontein-sludge), carbon growth substrates (yeast extract lactate, glucose medium and fruit cocktail effluent) and batch cultivation techniques (roller-table and shake-waterbath), on batch cultivation studies was also evaluated in terms of granule activity, ECP composition and granule formation. A physical extraction method was used to quantify the ECP content of UASB granules from six different sources. The optimal extraction time was taken as the time needed before cell lysis took place, and before intracellular material started contributing to the ECP content of the granules. It was concluded that the ECP composition was affected by the wastewater composition fed to the original UASB reactors, It was also found that the activity test results could be used to indirectly predict the activity of the different trophic groups present in the UASB granules. A correlation was found between the activity test results and the total ECP content, and this showed that the granules with the higher ECP yields exhibited greater biogas (SB) and methanogenic (SM) activities. However, based on the activity data and total ECP content, it appeared that a protein:carbohydrate ratio < 1 affected the activity of the granules, The sludge source used as inoculum for batch cultivation of anaerobic granular sludge had a significant effect on granule formation. The use of a pre- granulated raw anaerobic sludge, such as the Paarl-sludge, resulted in a greater increase in granule numbers at the end of the cultivation period. The acetic acid activity profiles showed that the acetoclastic methanogens that are involved in initiation of granulation by nucleus formation, were inactive or absent in the different batch systems, with the exception of the roller-table glucose cultivated Kraaifontein-sludge (RKG) batch system. The addition of glucose as carbon growth substrate for batch cultivation not only enhanced the activity of the acidogenic population, but also led to the establishment to a greater variety of granule trophic groups within all the glucose cultivated batch systems. The addition of fruit cocktail effluent as carbon substrate enhanced ECP production in the Paarl-sludge cultivated batch systems. However, the addition of carbon substrates showed no discernible trend on granule formation itself. The roller-table cultivation technique resulted in the higher increase in granule numbers, and it was speculated that the more vigorous shake-waterbath technique probably shortened the contact time between biomass and substrate. Large variations in the ECP composition of the different batch systems were found, and these were ascribed to the composition heterogeneity of different sludges. For future studies, it is advisable to characterise sludge, both chemically and microbiologically before using as inoculum. The selection of an appropriate sludge inoculum should then lead to optimisation of the granulation process. / AFRIKAANSE OPSOMMING: Die aanvangsperiode van "Upflow Anaerobic Sludge Bed" (UASB) bioreaktors kan noemenswaardig verminder word deur die tydsame granulasie proses te versnel deur die vooraf lot-kweking van anaërobe granulêre slyk waarmee 'n reaktor dus geïnokuleer kan word in plaas van rou anaërobe slyk. Die presiese meganisme van granulevorming is nog nie welbekend nie, maar daar word beweer dat ekstrasellulêre polimere (ECP) wel 'n kritiese rol speel in die granulasie proses. Inligting; oor die presiese rol van ECP is ook nog beperk, en tans word daar nog , geen universele standaard metode vir ECP-ekstraksie gebruik nie. Gevolglik moet resultate vanaf verskeie navorsers met groot omsigtigheid vergelyk word. Die doelwitte van hierdie studie was om 'n ECP ekstraksiemetode te evalueer deur die ekstraksietyd te optimiseer, en dan te korreleer met die ECP samestelling en metaboliese aktiwiteit van die UASB granules. Die inwerking van veranderinge in omgewingskondisies, soos slykbronne (Paarl- en Kraaifontein-slyk), koolstofbronne (gisekstrak-Iaktaat-, glukose-medium en vrugtekelkie-uitvloeisel) en lot-kwekingstegnieke (rol-tafel en skud-waterbad) op lot-kweking studies in terme van granule aktiwiteit, ECP-samestelling en granulevorming is ook ondersoek. 'n Fisiese ekstraksie metode is gebruik om die ECP-inhoud van UASB granules vanaf ses verskillende bronne te bepaal. Die optimale ekstraksietyd is geneem as die tyd benodig voordat sellise sal plaasvind en die intrasellulêre materiaal 'n bydrae sal lewer tot die ECP-inhoud van granules. Dit is afgelei dat die ECP-samestelling beïnvloed word deur die samestelling van die afvalwater wat vir die oorspronklike UASB bioreaktors gevoer is. Voorts is gevind dat die aktiwiteitstoets resultate indirek gebruik kan word vir die voorspelling van aktiwiteit van die verskillende trofiese groepe wat in die UASB granules teenwoordig is. 'n Korrelasie is gevind tussen die aktiwiteitstoets resultate en die totale ECP-inhoud wat aangedui het dat granules met hoër ECP opbrengste, beter biogas (SB) en metanogeniese (SM) aktiwiteit getoon het. Volgens die aktiwiteitsdata en totale ECP-inhoud het dit egter geblyk dat 'n proteïen:koolhidraat verhouding < 1 die aktiwiteit van granules beïnvloed het. Die slykbron wat as inokulum gebruik is vir lot-kweking van' anaërobiese granulêre slyk het 'n noemenswaardige invloed gehad op granulevorming. Die gebruik van 'n rou anaërobe slyk wat reeds 'n mate van granulasie getoon het, soos die Paarl-slyk, het 'n hoër toename in granule getalle aan die einde van die kwekingsperiode teweeg gebring. Die asynsuur aktiwiteitsprofiele het aangedui dat die asetoklastiese metanogene, wat hoofsaaklik betrokke is by inisiëring van granulasie deur kernvorming, onaktief of afwesig was in al die verskillende lotsisteme, met die uitsondering van die roltafel glukose-gekultiveerde Kraaifonteinslyk (RKG) lot-sisteem. Die toevoeging van glukose as koolstofbron vir lot-kweking het nie alleenlik die aktiwiteit van die asidogene populasie verhoog nie, maar het ook bygedra tot die vestiging van 'n groter verskeidenheid van granule trofiese groepe. Die toevoeging van vrugtekelkie-uitvloeisel as koolstofbron het die produksie van ECP verhoog in die Paarl-slyk gekweekte lot-sisteme. Die toevoeging van koolstofsubstrate het egter geen merkbare verandering getoon in granulasie opsigself nie. Die rol-tafel kwekingstegniek het 'n hoër toename in granule getalle tot gevolg gehad, en dit is gespekuleer dat die meer kragtige skud-waterbad tegniek waarskynlik die kontaktyd tussen die substraat en biomassa verkort het. Groot variasies is gevind in die ECP-samestelling van die verskillende lotsisteme, en dit is toegeskryf aan die heterogene samestelling van die verskillende slyke. Vir toekomstige navorsing is dit raadsaam om slyk voor gebruik as inokulum beide chemies en mikrobiologies te karakteriseer. Die keuse van 'n geskikte slyk-inokulum sal bydra tot die optimisering van die granulasie proses.

Thesis (Msc Food Sc (Food Science))--University of Stellenbosch, 2007. / The distillery industry generates large volumes of heavily polluted wastewater and thus effective wastewater treatment is essential. It has been reported that a chemical oxygen demand (COD) reduction of more than 90% can be achieved when wine distillery wastewater (WDWW) is treated in an upflow anaerobic sludge blanket (UASB) reactor. The first objective of this study was to investigate UASB treatment of WDWW and to try to enhance the efficiency by using ozonation treatments. Secondly, the impact of grain distillery wastewater (GDWW) on UASB granules was determined. The third objective was to determine whether ozonation and enzymatic treatment combinations might improve the biodegradability of GDWW and thus make GDWW more amenable to UASB treatment. It was found that UASB treatment combined with ozonation improved the WDWW treatment efficiency. When diluted WDWW (chemical oxygen demand COD = 4 000 mg.L-1) was ozonated (dose = 47 mg.L-1) in a 50 L venturi circulating contactor system, the COD reduction was 7%. When WDWW was treated in a laboratory-scale UASB reactor (substrate pH = 7.0, COD = 4 000 mg.L-1 and organic loading rate (OLR) = 4.0 kg COD.m-3.d-1), the COD reduction was 92%. When the UASB treatment was combined with either pre- or postozonation, the COD reduction was 94 and 96%, respectively. When UASB treatment was combined with pre- and post-ozonation, a COD reduction of 98% was achieved. The activity of the UASB granules was also found to improve over time, despite the addition of the ozonation treatment. It has been reported that operational problems occur when GDWW is treated in an UASB reactor as a result of the encapsulation of the granules. This was confirmed when granules from a full-scale UASB treating WDWW became encapsulated in a layer after being exposed to GDWW (COD = 4 000 mg.L-1) for 24 d. The results showed that the lipid content of the granules increased from 1.25 to 60.35 mg lipid.g-1 granule over the 24 d exposure period. Therefore, granules exposed to GDWW were encapsulated in a lipid-rich layer and as a result the contact between the GDWW and microbial consortium in the granules was reduced. The operational problems found during the industrial UASB treatment of GDWW were ascribed to the encapsulation of the granules. Combinations of ozonation (dose = 1 476 mg.L-1) generated in a 2 L bubble column and enzymatic treatments (1% FogFreeTM (FF) dosage and 2 d incubation at 35°C) were found to improve the biodegradability of GDWW. This improvement was in terms of lipid reduction in GDWW, granule activity and visual appearance of the encapsulating layer of the granules. The highest lipid reduction (90%), highest granule activity, lowest lipid content of the granules (3.74 ± 0.10 mg.g-1 granule) and best visual appearance were achieved in ozonated GDWW treated with 1% FF, followed by just ozonation. The higher lipid reduction and subsequent higher granule activity were ascribed to the reduction in lipids which resulted in the fact that fewer lipids were available to encapsulate the granules. As a result of the lipid reduction, the granule activity improved and the GDWW was made more amenable to UASB treatment. This study proved that UASB treatment combined with ozonation led to an enhancement of the treatment efficiency of WDWW. It was also found that the cause of the operational problems during UASB treatment of GDWW was as a result of the granules being encapsulated in a lipid-rich layer. It was established that treating GDWW prior to UASB treatment improved the biodegradability of GDWW. The data from the study showed that high lipid reduction in the GDWW directly led to better granule activity, lower granule lipid content and a thinner encapsulating layer. Based on the data from this study, it is recommended that GDWW be ozonated prior to other treatments because it can be done inline and the costs would be lower than that of enzymatic treatments.

The presence of heavy metal ions in waste streams is one of the most pervasive environmental issues of present times. A rotating biological contactor (RBC) was used to investigate the potential capacity of microbial biofilms in remediation of the metal ion species from a mixed metal contaminated effluent solution containing Cr+3 , Pb+2 and Cu+2 , each at a concentration of 200 mg r1 • In the first part of this study the effectiveness of various support materials for the development of microbial biofilms capable of removing heavy metals from a synthetic effluent was investigated. EDX analysis showed that none of the support matrices investigated, viz. gravel, polyester batting and sand, adsorbed metal ions on their surfaces; hence, metal adsorption was due purely to microbial activities. The biofilms attached more firmly and uniformly to polyester batting than to gravel and sand. The characteristics of polyester batting which made it a superior support matrix were its surface roughness and porous hydrophilic nature, which provided a larger surface area for the adhesion of microorganisms and attraction of nutrients during the biofilm development process. The selective accumulation of metal ion specIes by various microbial populations grown as biofilm using polyester batting as support matrix in separate compartments of a single-stage RBC bioreactor was examined. Lead ions were readily accumulated by almost all the microbial biofilms tested. Fungus-dominated biofilms selectively accumulated chromium ions whereas biofilms comprising mainly bacteria more readily accumulated copper ions from the mixed metal contaminated effluent solution. However, where interactions between the bacterial and fungal components were encouraged the mechanical stability of the biofilms was enhanced so that large amounts of all three metal ion species were removed by this biofilm. The combined effect of a series of bench-scale columns containing liquid humic acid and a three stage RBC bioreactor on the removal of metal ion species from a mixed metal contaminated effluent was investigated. After seven days of treatment the combined system had removed approximately 99% of the Cr+3, 98% of the Pb+2 and 90% of the Cu+2 ions from the mixed metal contaminated synthetic effluent. Complexation of the metal ions with humic acid was the predominant factor accounting for approximately 68-86% Cr+3 , 70-86% Pb+2 and 53-73% Cu+2 removal levels within the columns. A large proportion of the remaining Cr+3 and Pb+2, but not of the Cu+2, was removed in compartment 1 of the RBC. This suggested that the presence of the former two metals in solution might have reduced the removal of the Cu+2 ions from the system. The removal of substantially large amounts of the competing ions chromium and lead during the initial stages of the treatment process meant that copper was successfully taken up in the second and third RBC compartments. Hence, the economy of the treatment process was improved as larger quantities of the metal ions were removed in a shorter period of time than was possible when using the individual treatments (humic acid-metal complexation and biofilm adsorption) separately. More than 75%,92% and 86% of the adsorbed Cr+3 , Pb+2 and Cu+2 ions, respectively, were recovered from the three RBC bioreactor compartments following repeated washing of the biofilms with 0.1 M HCI. This relatively easy desorption suggested that the metal ions were simply adsorbed onto the surfaces of the biofilm cells rather than being taken into the cytoplasm of the cells. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

Thesis submitted in the requirements for the degree Doctor Technologiae: Chemical Engineering in the Faculty of Engineering at the CAPE PENINSULA UNIVERSITY OF TECHNOLOGY, 2013 / To date, limited information has been published on textile wastewater treatment, for re-use, in South Africa (SA), with treatment processes focusing on conventional wastewater treatment methods. A large contributor to the contamination of water within textile industries is from dyehouse processes. A major concern in textile wastewater treatment is the release of azo dyes and their metabolites, some of which are carcinogenic and mutanogenic, into the environment since they are xenobiotic and aerobically recalcitrant to biodegradation. A necessity therefore exists to find an effective treatment method capable of removing both the strong colour and the toxic organic compounds from textile wastewater. Membrane bioreactors (MBRs) are favoured when treating high-strength wastewater, since the membrane area is determined by the hydraulic throughput and not the biological load; no sludge is wasted and all bacteria are retained within the reactor, including specific bacteria capable of degrading the toxic, non-biodegradable constituents present in textile wastewater. MBR systems, using various configurations have been utilised extensively in the rest of the world to treat textile wastewater at both lab and pilot-scale. This DTech project formed part of a collaborative Water Research Commission (WRC) funded project K5/1900 - Pilot application of a dual-stage membrane bioreactor (dsMBR) for industrial wastewater treatment. The main purpose of this study was the on-site evaluation of a pilot-scale dsMBR incorporating two ultrafiltration (UF) sidestream membrane modules for the treatment, recovery and re-use of textile wastewater. The objectives of this project were to determine the treatment efficiency of the system; to evaluate the degree of colour removal from the textile wastewater; to improve residual colour removal within the system using treatment processes, such as NF and RO, as well as to propose a design and cost for a full-scale plant. A textile industry located in Bellville, Western Cape, was chosen as the industrial partner for the on-site evaluation of a semi-automated pilot wastewater treatment MBR plant using two 5.1 m2 Norit X-flow AirliftTM membrane modules. Since the wastewater treatment system was located on the premises, real continuously changing industrial wastewater was being treated. The industrial textile wastewater was treated in a series of tanks: 1) an anaerobic tank, which cleaved the azo bonds of the reactive dyes; 2) an anoxic tank containing reduced amounts of dissolved oxygen, in which denitrification occurred; and 3) an aerobic tank, in which i) nitrification, as well as ii) mineralisation of the aromatic amines occurred. The UF-membrane modules would account for the removal of any organic material. The wastewater stream was characterised by a chemical oxygen demand (COD) range of between 45 to 2,820 mg/L and an average biological oxygen demand (BOD) of 192.5 mg/L. The dsMBR achieved an average COD reduction of 75% with a maximum of 97% over the 220 day test period. The COD concentration obtained after dsMBR treatment averaged at 191 mg/L, which was well within the City of Cape Town industrial wastewater discharge standard. The average reduction in turbidity and TSS was 94% and 19.6%, respectively, during the UF-MBR stage of the system. Subsequent treatment of the UF permeate with nanofiltration (NF) for 4 days, alternated with reverse osmosis (RO) for 14 days removed both the residual colour and salt present in the UF permeate. A consistent reduction in the colour of the incoming wastewater was evident. The colour in the wastewater was reduced from an average of 659 ADMI units to ~12 ADMI units in the NF permeate, a lower American dye manufacturing index (ADMI) (i.e. method of colour representation) compared to the potable water (~17 ADMI units) utilised by the industrial partner in their dyeing processes. The colour was reduced from an average of 659 to ~20 ADMI units in the RO permeate, a lower ADMI and therefore colour when compared to the potable water. An average conductivity rejection of 91% was achieved with conductivity being reduced from an average of 7,700 to 693 μS/cm and the TDS reduced from an average of 5,700 to 473 mg/L, which facilitated an average TDS rejection of 92%. Based on the composition of the UF permeate fed to the RO membrane a maximum removal of 98.7% was achieved for both conductivity and total dissolved solids (TDS). The proposed full-scale plant would incorporate a UF-MBR system, followed by NF, RO, flocculation and a filter press. Therefore, the two waste products produced during operation of the proposed full-scale plant, would be the solid filter cakes and the liquid filtrate from the filter press. Implementing the proposed full-scale plant it would cost the industrial partner an operating cost of ZAR 113.85 and ZAR 3,415.49 to treat 97.1 m3 and 2,913 m3 of textile wastewater, respectively, per day and per month. This results in an annual saving of ZAR 845,848 on potable water expenses. This research, would provide SA textile industries, with an option to: 1) reduce their water consumption, thereby utilising less of a valuable decreasing commodity; 2) meet the SA government discharge standards and reduce their discharge costs; 3) reduce their carbon footprint (i.e. reduce their impact on the environment) by re-using their treated wastewater and therefore using less water from the municipality; and 4) decrease their annual expenditure on water, since the treated wastewater would be available for re-use.