Este trabalho apresenta e discute os resultados obtidos na operação de um reator anaeróbico com manta de lodo (UASB) de 10,5 L, alimentado continuamente com substrato sintético (DQO = 2000 mg/L) e submetido ao aumento progressivo da concentração de sulfato (de 25 a 10000 mg \'SO IND.4\'POT.2-\'/L). A vazão de 16 L/dia, corresponde ao tempo de detenção hidráulico (e) de 15,6 h foi mantida constante durante os 10 meses do experimento. O monitoramento da operação do reator incluiu a determinação dos principais parâmetros do processo, visando detectar a inibição da metanogênese pelas bactérias redutoras de sulfato (BRS) que competem, com as metanogênicas, pelos mesmos substratos. Constatou-se estimulação do processo até a concentração de 400 mg \'SO IND.4\'POT.2-\'/L, a partir da qual ocorreram sequencias de quedas de eficiência seguidas de recuperação do reator, mantendo sempre a eficiência média de remoção de DQO acima de 80%. Embora a atividade metanogênica tenha sido parcialmente afetada, os balanços de \'SO IND.4\'POT.2-\' mostraram que a redução do sulfato não foi o principal responsável pela inibição ocorrida. / This thesis presents and discuss the results obtained during the operation of a 10.5 L upflow sludge blanket reactor, continuously fed with synthetic substrate (COD = 2,000 mg/L), which was subjected to step-increase of sulfate concentrations ranging from 25 to 10,000 mg \'SO IND.4\'POT.2-\'/L. The flow-rate of 16 L/d corresponding to a hydraulic detention of 15.6 h, was kept constant throughout the 10 months of experimentation. Operation monitoring included the determination of the main process parameters, aiming to detected the inhibition of lhe methanogenesis by the sulfate reducing bacteria (SRB) which autocompete the methanogens in the presence of common substrates. Stimulation effects were obseved up to the sulfate concentration of the 400 mg.\'SO IND.4\'POT.2-\'/L. Further on, sequences of falls of efficiency followed by restoration of the reactor performance defined the behavior pattern of the reactor, which maintained an average COD removal efficiency higher than 80%. Although the methanogenic activity has been parcially affected, \'SO IND.4\'POT2-\' balances showed that the sulfate reduction activity was not the main process responsible for such inhibition.

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2005. / The Up-flow Anaerobic Sludge Blanket Reactor (UASB) provides a state-of–the-art solution to effluent treatment by anaerobic digestion: sludge production is dramatically lower than in other digestion processes, and energy is gained from the system if the produced biogas is converted to electricity and/or heat. The UASB is a modified fluidised bed reactor, with the solid state ‘catalyst’ being granulated anaerobic sludge, and the liquid phase the effluent that needs to be treated. A gas cap is installed to serve as a carbon dioxide and methane collector. This biogas (carbon dioxide and methane) is produced by the stepwise decomposition of complex carbohydrates and proteins via a consortium of micro-organisms living in a symbiotic environment known as a granule. A typical UASB reactor has an organic removal rate of 89-93% Chemical Oxygen Demand (COD) and operates optimally at loadings of 9.8-11 kg COD/ m3 reactor volume/day. Unfortunately, one major problem hampers the efficiency of this reactor to such an extent that the unit is only economically viable in exceptional cases; if the reactor is inoculated with un-granulated anaerobic sludge, start-up times of up to 12 months can be expected. The lengthy start-up times motivated the search for an artificial way to cultivate USAB granules. Early research (done on lab-scale, 400ml vessel volumes) proved that, under a specified set of environmental conditions, granule growth can occur in an artificial environment. Yet these laboratory-scale vessels did not facilitate scale-up or the study thereof. This led to the main problem statement of this research project: namely to design, commission, and optimise benchscale bioreactors that will generate granulated anaerobic sludge in an incubation period of 20 days. These units should also facilitate in the determining of parameters that will assist in the design of a scale-up to a UASB granule producing reactor of economically viable size. Two bench-scale reactors were initially designed specifically to “mimic” the motion found in the laboratory-scale vessels. The results from these initial reactors proved that granulation cannot only be enhanced, but granules can actually be cultivated from dispersed anaerobic sludge in a larger artificial environment over an incubation period of only 20 days. The results were still far from satisfactory, as the granules produced were irregular in shape and the yield of usable granules (2.2 kg/m3 reactor volume) insufficient. A third test reactor was designed to “mimic” roller table movement and baffles were included. These results were much better and the yield was 4.4 kg/m3 reactor volume at a baffle tipspeed of 0.0055 m/s. The optimisation was extended further to include the inoculation sludge and the feed medium. A C:N:P ratio of 10:1:4 proved to yield the best results. Monovalent anions, hydrogen concentration and a pH-level outside the 6.5 to 7.2 range evidently had an inhibitory effect on the granulation rate. After the optimisation study the third test unit produced a usable granule yield of 15.2 kg/m3 reactor volume over the 20-day incubation period. The incubation period can be separated into 3 distinct phases, namely the acidification, stabilisation and growth phases. From the mass balance it was found that most of the COD and nutrients were used for ECP production in the acidification phase. During the stabilisation phase, the COD and nutrients were mostly used for nucleus formation, and finally in the growth phase the COD was used for granule growth. To study the effect the internal surface area of the reactor has on the granulation process, 3 scale-down versions of the third test unit were constructed. Within the studied range, a yield of usable granules of 40 kg/m2 reactor internal surface area was obtained.

Thesis (PhD (Food Science))--University of Stellenbosch, 2009. / In the absence of anaerobic granules, anaerobically digested sewage sludge is frequently used to seed industrial upflow anaerobic sludge blanket (UASB) reactors. Because of its flocculent nature, start-up with digested sludge instead of granular sludge proceeds much slower and presents various operational problems. Any manner in which the granulation of digested sludge can be enhanced would benefit UASB reactor start-up and application in developing countries such as South Africa. The main objective of this dissertation was to improve granulation and reduce UASB reactor start-up by using pre-treated digested sludge as seed. The sludge was pre-treated based on the batch granulation-enhancement model of Britz et al. (2002). The main aim of the model was to improve extracellular polymer (ECP) production of lactate-utilising populations by applying short-term controlled organic overloading in a mechanically agitated environment. The batch granulation-enhancement (pre-treatment) process was applied to an ECP-producing digester strain, Propionibacterium jensenii S1. Non-methanogenic aggregates were formed when batch units were incubated on a roller-table instead of a linear-shake platform. Larger, more stable aggregates were obtained in the presence of apricot effluent medium. Preliminary batch granulation-enhancement studies confirmed that using the roller-table as mixing system had a positive influence on batch granulation-enhancement. The roller-table showed the most potential for handling larger volumes in comparison to a linear-shake waterbath and linear-shake platform. The addition of 450 mg.L-1 Fe2+ at the start of the study also influenced aggregate numbers positively. These studies revealed that pre-treatment results varied depending on the seed sludge source. A denaturing gradient gel electrophoresis (DGGE) method was applied for the detection of Archaea in digested sludges and UASB granules. In addition, a methanogenic marker containing methanogens important to the granulation process was constructed to aid identification. The positive influence of DMSO and “touchdown” PCR on the elimination of artifactual double bands in DGGE fingerprints were also demonstrated. Results revealed that only one of the four digested sludges tested contained Methanosaeta concilii (critical to granular nuclei formation) while it was present in all the UASB granules regardless of substrate type. Four digested sludges were obtained from stable secondary digesters. DGGE indicated the presence of M. concilii in all sludges. The Athlone 4Sb-sludge was the only sludge which exhibited measurable methanogenic activity during substrate dependent activity testing. The ST-sludge showed the highest increase in volatile suspended solids (VSS) particles ≥0.25 mm2. Laboratory-scale UASB reactor start-up was done with both sludges and start-up proceeded better in the Athlone 4Sb-reactor. Athlone 4Sb-sludge batches were pre-treated in a rolling-batch reactor in the presence of either lactate or sucrose and used to seed lab-scale UASB reactors B (sucrose seed) and C (lactate seed). Start-up efficiencies were compared to a control (Reactor A). Overall Reactor B was more efficient that the control. At the end of the study the Reactor B sludge had a higher methanogenic activity than the control reactor. It also had the highest increase in VSS ≥1.0 mm2. Pre-treatment of digested sludge in the presence of sucrose, therefore, aided granulation and reduced UASB reactor start-up time.

Water scarcity is a global concern that impacts many developing countries, forcing people to depend on unclean water sources for domestic, agricultural, and industrial needs. Wastewater is an alternative water source that contains nutrients needed for crop growth. Wastewater reuse for agriculture can cause public health problems because of human exposure to pathogens. Pathogen monitoring is essential to evaluate the compliance of wastewater with established World Health Organization (WHO) and U.S. Environmental Protection Agency (EPA) wastewater reuse guidelines. Indicator organisms are commonly used to detect pathogens in water and wastewater because they are quick and easy to measure, non-pathogenic, and have simple and inexpensive methods of detection. The objective of this research was to develop a modified double agar layer assay method that can be conducted in the field to quantify bacteriophage to assess the quality of wastewater for agricultural reuse. Results from the modified double agar layer assay were used to investigate the potential of somatic coliphage as an indicator organism for assessing the potential presence of enteric viruses in developing world treated wastewater, and to use the criteria of a good indicator organism to compare the potential of two commonly used indicator organisms, somatic coliphage and fecal coliforms, as an indicator of enteric viruses in wastewater. A modified EPA double agar layer method was developed and deployed in a developing world rural community to effectively quantify the concentration of somatic coliphage in a community managed wastewater treatment system composed of a Upflow Anaerobic Sludge Blanket (UASB) reactor followed by two maturation lagoons. The modified method served as a good indicator of enteric viruses in the water. Somatic coliphages were easily detected and quantified in the field setting using a modified double agar layer method. Somatic coliphage was found to be a potential indicator for enteric viruses rather than fecal coliforms because of their similarity in characteristics and resistances to wastewater treatment. The concentration of somatic coliphage was only reduced by 1.05 log units across the two series maturation lagoon system. Previous literature suggested removal would range from 2.1 to 4.6 log units. Influent wastewater (previously treated by an UASB reactor) had a concentration of 4.38 E+06 PFU/ 100 mL (standard deviation = ±3.7E+06, n = 9) and the treated effluent contained 3.90 E+05 PFU/100 mL (standard deviation = ± 4.5E+05, n = 8) of somatic coliphages. Results suggest that somatic coliphage is a good potential indicator for enteric viruses in wastewater but further research needs to be done.

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, 2004. / ENGLISH ABSTRACT: The wine industry significantly contributes to South Africa's water demand and subsequent pollution of the limited resource. Wastewater is produced throughout the year with an increase in volume and organic load during the vintage season. Anaerobic digestion (AD), specifically the upflow anaerobic sludge bed (UASB) technology has been shown to be feasible in the treatment of cellar wastewater. However, the legal standard for chemical oxygen demand (COD) for disposal in a natural water resource (75 rnq.L") is often not met. The aim of the study was to conduct a laboratory-scale investigation into the feasibility of combining pre- and post-ozonation processes with AD in order to achieve a final COD closer to the legal disposal limit. While acclimatising an UASB bioreactor containing mixed anaerobic granules to a cellar wastewater with a pH set at 8.0, stable-state conditions were not reached. Sucrose additions to the substrate, increased substrate loads, heattreatment of the substrate and an addition of isolated cellar effluent bacteria to facilitate degradation prior to AD, were all unsuccessful in maintaining stable-state in terms of COD removal efficiency. Once the substrate pH was re-set to 7.5, the reactor stabilised. The lowest efficient operational pH was found to be 5.73 resulting in a COD removal of 88% at a substrate COD < 5 000 rnq.L". At a substrate pH of 6.0, the lowest efficient operational hydraulic retention time (HRT) and corresponding organic loading rate (OLR) were 19.7 hand 9.75 kg COD.m-3d-1 , respectively, with the COD removal being maintained around 84%. The reactor effluent still had a final COD of 1280 rnq.L", which was well above the legal South African limit. Dominant bacteria were isolated from raw cellar wastewater and identified as Acinetobacter haemolyticus, Burkholderia cepacia and Cryseomonas luteola. In order to investigate the possibility that ozonation improved biodegradability, the growth of the isolates at 35°C was monitored over 24 h in sterile ozonated and non-ozonated substrates from the vintage and non-vintage seasons. All the isolates increased by at least 1.5 log cycles in the control substrates from both seasons. Ozonation of the wastewater batches for 10 min at a rate of 73 rnq.L" led to slightly increased growth of the inoculants in the substrate batch from the vintage season. For the substrates from the non-vintage season, ozonation had an inhibitory effect on the bacterial growth. A 5 min ozonation treatment at a concentration of 73 rnq.L" was found to be optimal for both a pre- and post-treatment to UASB-treatment of cellar wastewater. Both UASB treatment and ozonation were effective in reducing the COD by 85% and 20%, respectively. The COD reduction was improved to 88% when UASB treatment was combined with post-ozonation. The total reduction in total suspended solids (TSS) for the combined process was 97%, compared to 80% for UASB and 73% for an ozone treatment alone. The reduction for volatile suspended solids (VSS) was 98% compared to 81% for UASB and 73% for the ozone treatment alone. The total reduction when using a pre-ozonation UASB treatment combination was an average of 86% for COD. The TSS and VSS were both reduced by 95%. Biogas production increased from 1.4 L.d-1 to 3.8 L.d-1 when an ozonated wastewater was used as substrate. When the UASB treatment was combined with both a pre- and post-ozonation treatment process, the COD was reduced by 89% while TSS and VSS were both reduced by 99%. This study showed that pre- and post-ozonation treatment processes could successfully be utilised to improve UASB treatment of cellar wastewater. Although the legal limits for discarding into a natural resource were not met, significant progress was made in reducing COD levels. Cellar wastewaters do however, vary according to season and the wastewater composition could affect the efficiency of a pre-ozonation process. / AFRIKAANSE OPSOMMING: Die wynindustrie maak "n beduidende bydrae tot die eise wat aan Suid-Afrika se waterbronne gestel word en gevolglik die besoedeling van die beperkte hulpbron. Afloopwater, wat in volume en organiese lading gedurende die parstyd toeneem, word reg deur die jaar opgelewer. Anaërobiese vertering (AV), spesifiek die "Upflow anaerobic sludge blanket" (UASB) tegnologie, is alreeds suksesvol gebruik om kelderafloop te behandel. Die wetlike vereiste vir chemiese suurstof behoefte (CSB) vir storting in "n natuurlike hulpbron (75 rnq.L"), word egter dikwels nie bereik nie. Die doel van die studie was om in "n laboratorium-skaal ondersoek AV te kombineer met voor- en na-osoneringsprossesse, om sodoende te poog om "n CSB nader aan die wetlike standaard te verkry. Terwyl"n UASB bioreaktor wat gemengde anaerobiese granules bevat het, geakklimatiseer is tot kelderafloop met "n pH gestel tot 8.0, kon stabiele toestande nie bereik word nie. Die byvoeging van sukrose tot die substraat, verhoogde substraatladings, hitte-behandeling van die substraat en die byvoeging van geïsoleerde kelderafloop bakterië om substraatafbraak voor AV aan te help, was onsuksesvol om stabiliteit in terme van CSB-verwydering, te handhaaf. "n Verstelling van die substraat pH na 7.5, het gelei tot reaktorstabiliteit. By die laagste doeltreffende bedryfs-pH van 5.73 en substraat CSB < 5 000 rnq.L", was die CSB-verwydering 88%. By "n substraat pH van 6.0 was die laagste doeltreffende bedryfs-hidroliese retensie tyd en -organiese ladingstempo 19.7 h en 9.75 kg CSB.m-3d-1, onderskeidelik, terwyl die CSB verwydering rondom 84% gehandhaaf is. Die CSB van die reaktoruitvloesel van 1 280 rnq.L", was steeds ver bo die wetlike vereiste. Dominante bakterië is uit kelderafloop geïsoleer en as Acinetobacter haemolyticus, Burkholderia cepacia en Cryseomonas luteola, geïdentifiseer. Die moontlikheid dat osonering bioafbreekbaarheid bevorder, is ondersoek deur die groei van die isolate by 35°C oor 24 h in steriele geësoneerde en ongeësoneerde substrate te monitor. Die substrate is berei vanaf kelderafloop wat in die parsseisoen sowel as die nie-parsseisoen versamel is. AI die isolate het met ten minste 1.5 log siklusse in die kontrole substrate van beide seisoene, vermeerder. Vir die kelderafloop wat in die parsseisoen versamel is, het osonering vir 10 min teen 73 rnq.L" gelei tot effens verbeterde groei van die innokulante. Osonering het 'n onderdrukkende effek op die groei van bakterië in die afloopwater versamel in die nie-parsseisoen, gehad. Osonering vir 5 min teen 'n konsentrasie van 73 rnq.L" is as optimum vir beide voor- en na-osoneringsbehandeling tot UASB-behandeling van die kelderafloop, gevind. UASB-behandeling en osonering het die CSB met 85 en 20% onderskeidelik, verminder. Die vermindering kon tot 88% verhoog word wanneer UASB-behandeling met na-osonering gekombineer is. Die vermindering in totale gesuspendeerde vastestowwe (TGV) vir die gekombineerde proses was 97%, in vergelyking met 80% vir UASB- en 73% vir osoonbehandeling alleen. Die vermindering in vlugtige gesuspendeerde vastestowwe (VGV) was 98% in vergelyking met 81% vir UASB- en 73% vir osoonbehandeling alleen. Die totale CSB verwydering vir 'n voor-osonerings UASB kombinasie was gemiddeld 86%. Die TGV en VGV is beide met 95% verminder. Biogasproduksie het ook vermeerder vanaf 1.4 L.d-1 tot 3.8 L.d-1 toe geosoneerde afloopwater as substraat gebruik is. Die kombinasie van UASB-behandeling met voor-osonering, sowel as na-osonering het gelei tot 'n CSB-verwydering van 89% terwyl TGV en VGV beide met 99% verminder is. Hierdie studie het getoon dat voor- en na-osonering suksesvol gebruik kan word om UASB-behandeling van kelderafloop te verbeter. Hoewel wetlike vereistes vir storting in 'n natuurlike hulpbron nie bereik is nie, is beduidende vordering gemaak in die verlaging van CSB-vlakke. Die verskil in die samestelling van kelderafloop gedurende die onderskeie seisoene, kan egter die doeltreffendheid van die voor-osoneringsproses beïnvloed.

Orientador: Roberto Alves de Oliveira / Banca: Wanderley José de Melo / Banca: Marco Antonio Penalva Reali / Resumo: Nas estações de tratamento de água (ETA), as impurezas retiradas da água juntamente com os produtos hidrolisados dos coagulantes dão origem aos resíduos dos decantadores e das águas de lavagem dos filtros, denominados lodo de ETA. Neste experimento, foi avaliado a disposição de lodo de ETA em reatores anaeróbios de fluxo ascendente com manta de lodo (UASB), com volume de 18 L e 38 L, visando o tratamento para destinação adequada do efluente e do lodo excedente dos reatores. Tal experimento foi dividido em quatro ensaios, com tempo de detenção hidráulica de 24 h nos reatores UASB, previamente inoculados com lodo granulado e floculento. Foram observadas condições em que foram aplicadas ao reator UASB, lodo de ETA, com DQOtotal de 161 mg/L e ST de 1.948 mg/L, e misturas de lodo de ETA com esgoto sanitário nas seguintes proporções, em volume, de 1:1 (lodo de ETA : esgoto sanitário) com DQOtotal de 458 mg/L e ST de 3.039 mg/L; 1:3, com DQOtotal de 519 mg/L e ST de 1.562 mg/L e 1:7, com DQOtotal de 1.174 mg/L e ST de 3.502 mg/L. Os valores médios de eficiência de remoção da DQOtotal foram de 75 a 83% e de ST de 73 a 87%. A maior remoção de P foi de 90% e de N de 41%. As eficiências de remoção de Ca, Co, Fe, Mn, Ni, Cu, Pb, Cr e Zn foram superiores a 70%, confirmando a alta capacidade de bioadsorção de metais do lodo anaeróbio. As concentrações de N-am, Fe, Mn, Ni, Cd, Pb, Cu, Cr e Zn atenderam aos padrões de lançamento de efluentes da legislação federal. Ocorreram remoções de E. coli acima de 90%, resultando contagens do efluente de 1,5x10 UFC/100 mL, apresentando qualidade microbiológica correspondente à água doce de classe 1, conforme os padrões estabelecidos na Resolução CONAMA 357/2005 . O lodo do reator UASB apresentou valores de relação SV/ST e E. coli que permitiram o seu uso na agricultura. / Abstract: In water treatment plants (WTP), impurities removed from water mixed with coagulative hydrolytic product create waste of decanters and launderings water filters residues called WTP sludge. In this experiment the WTP sludge disposal in an Upflow Anaerobic Sludge Blanket Reactor (UASB) were evaluated, with a volume of 18 L and 38 L, aiming treatment for appropriate disposal of effluent and excess sludge reactors. Such experiment was divided into four assays, with hydraulic detention time of 24 h in UASB reactors by being inoculated previously with flocculent and granular sludge. It was observed conditions in wich it was added to the UASB reactor WTP sludge with CODtotal of 161 mg/L and TS of 1,948 mg/L, and mixtures of WTP sludge and wastewater in the following amounts, in volume, 1:1 (WTP sludge : wastewater) with CODtotal of 458 mg/L and TS of 3,039 mg/L; 1:3 with CODtotal of 519 mg/L and TS of 3,382 mg/L and 1:7 with CODtotal of 1,174 mg/L and TS 3,501 mg/L. The average values of removal efficiency of CODtotal were 75 the 83% and TS 73 the 87%. The highest P removal was 90% and the highest one of N was 41%. Removal efficiencies of Ca, Co, Fe, Mn, Ni, Cu, Pb, Cr and Zn were higher than 70% wich confirmed the high capacity of biosorption and bioaccumulation of metals from anaerobic sludge. Concentrations of Nam, Fe, Mn, Ni, Cd, Pb, Cu, Cr and Zn fit the quality standards of effluent discharge required by federal legislation. There were removals of E. coli higher than 90% which resulted in counts for effluent of 1.5x10 CFU/100 mL and it showed microbiological quality corresponding to freshwater class 1, according to the standards established in CONAMA Resolution 357/2005. Sludge from the reactor showed ratios of VS /TS and E. coli which allows agricultural use. / Mestre

Auf Grund der Zielstellungen des Klimaschutzes und den Szenarien der Ressourcenverfügbarkeit sowie der Energiekostenentwicklung wird das Energiemanagement bei der Betriebsführung von Abwasser- und Schlammbehandlungsanlagen an Bedeutung gewinnen, wobei die Maßnahmen des Energiemanagements mit den grundlegenden Zielen der Abwasser- und Schlammbehandlung zu vereinen sind. Zentrale Elemente des Energie-managements stellen die Nutzung eines breiten Energiespektrums und der Einsatz energieeffizienter Betriebsführungen, Techniken und Technologien dar. Eine Möglichkeit betriebstechnischer Optimierungen besteht in der Anwendung der Verfahrenskombination aus Desintegration und anaerober Schlammstabilisierung. Seit 10 – 15 Jahren wurde intensiv auf dem Gebiet der Klärschlammdesintegration geforscht, ohne dass allgemeingültige Aussagen über die Möglichkeiten der Desintegration ermittelt wurden. Anlässlich der dargestellten Sachlage und eigener Forschungsarbeiten auf dem Gebiet der Klärschlammdesintegration entstand die Idee zur vorliegenden Arbeit. Die Arbeit gibt in einer Literaturrecherche einen umfangreichen Überblick vorhandener Untersuchungen zur Verfahrenskombination Klärschlammdesintegration und anaerobe mesophile Schlammstabilisierung. Zu den untersuchten Desintegrationsapparaten und techniken gehören Rührwerkskugelmühle, Ultraschallhomogenisator, Hochdruck-homogenisator, Lysatzentrifuge, thermische Vorbehandlung, Hochleistungspulstechnik, Ozonbehandlung und die Kombination von thermischer Vorbehandlung mit Säuren- bzw. Laugenaufschluss. Zur Verbesserung der für die Auswertungen benötigten Datenbasis wurden labortechnische Untersuchungen zur anaerob mesophilen bzw. zur anaerob thermophilen Überschussschlammstabilisierung sowie zur Verfahrenskombination von Überschussschlammdesintegration mit anaerob mesophiler bzw. anaerob thermophiler Schlammstabilisierung durchgeführt Zur Desintegration wurden eine Rührwerkskugelmühle PE 075 (Fa. Netzsch-Feinmahltechnik), eine Fliehkraftkugelmühle S 100 (Retsch GmbH), ein Ultraschallhomogenisator UP 400S (Fa. Dr. Hielscher GmbH) eingesetzt sowie ein thermischer Aufschluss bei 75 °C und eine Kombination von thermischem Aufschluss bei 75 °C mit anschließender Unterdruckentspannung durchgeführt. Die vorliegende Arbeit beinhaltet eine umfangreiche Bewertung der Verfahrenskombination Überschussschlammdesintegration mit anschließender anaerob mesophiler Schlammstabilisierung. Speziell wurden in diesem Zusammenhang gezielte Untersuchungen zu variierenden Klärschlammqualitäten, Milieubedingungen der anaeroben Stabilisierung (Verweilzeit, Temperatur) und Desintegrationsverfahren (mechanisch, niederthermisch) durchgeführt. Die Auswertungen zeigen, dass die Wirkung dieser Verfahrenskombination maßgeblich durch die anaerobe Abbaubarkeit der Referenzüberschussschlämme bestimmt wird, wobei sich der relative Einfluss der Desintegration mit steigendem Stabilisierungsgrad der Überschussschlämme erhöht. Als Grenze des organischen Abbauverhaltens von desintegrierten Überschussschlämmen kann von einem GV Abbau 59 % bzw. von einem zufuhrspezifischen Biogasanfall von 467 Nl/kg GVZu ausgegangen werden. Die aus der Desintegration resultierende Steigerung des anaeroben Abbauverhaltens von Überschussschlämmen bedingt einen Anstieg der CSBmf- bzw. NH4-N-Prozesswasserbelastung. Für die PO4-P- bzw.- Pges,mf-Anteile des Prozesswassers war kein Zusammenhang nachweisbar. Für die Verfahrenskombination Überschussschlammdesintegration mit anschließender anaerob mesophiler Schlammstabilisierung werden neben dem Einsatz zur Verbesserung des anaeroben Abbauverhaltens von Überschussschlämmen Anwendungsmöglichkeiten zur Sanierung überlasteter Faulbehälter bzw. zur Minimierung notwendiger anaerober Reaktionsvolumina aufgezeigt. Entsprechend der Ergebnisse dieser Arbeit ist durch eine vorgeschaltete Überschussschlammdesintegration eine Verkürzung der anaeroben Stabilisierungszeit von 16 – 24 d auf 7 – 12 d möglich, ohne dass eine Verminderung des organischen Abbaus von Überschussschlämmen toleriert werden muss. Gesonderte Auswertungen befassen sich mit der Wirkung der Desintegration auf die anaerobe Abbaubarkeit der im Überschussschlamm enthaltenen Stoffgruppen Fette, Kohlenhydrate und Eiweiße. Es ist festzustellen, dass das anaerob mesophile Abbauverhalten einzelner im Überschussschlamm gebundener Stoffgruppen infolge einer Vorbehandlung durch Desintegrationsverfahren nicht/oder nur unwesentlich verändert wird. Im weiteren Verlauf der Arbeit wurden Nomogramme zur Prognose des Desintegrations-einflusses auf das anaerob mesophile Abbauverhalten von Überschussschlämmen erarbeitet, die es dem planenden Ingenieur bzw. dem Betriebsingenieur gestatten, die Auswirkungen eines Einsatzes der Kombination von Überschussschlammdesintegration und anaerob mesophiler Stabilisierung zu bewerten. Die Darstellungen ermöglichen eine Spezifizierung nach dem Grad der mechanischen Abwasserbehandlung und des in der biologischen Abwasserbehandlung praktizierten Gesamtschlammalters. Die Prognosemöglichkeiten umfassen den für desintegrierte Überschussschlämme bei der anaerob mesophilen Schlamm-stabilisierung auftretenden GV Abbau, den zufuhrspezifischen Biogasanfall sowie die resultierende CSBmf- und NH4-N-Prozesswasserbelastung. In Abhängigkeit der Verfahrens- und Betriebsführung der Abwasserbehandlung führt die Desintegration im Mittel zu einer maximalen absoluten Veränderung des GV-Abbaus bzw. des zufuhrspezifischen Biogas¬anfalls von Überschussschlämmen von 13 % bzw. 90 l/kg GVZu. Die Spannweiten der CSBmf- bzw. NH4-N-Prozesswasserbelastung betragen 0,5 – 6,5 % bzw. 3,0 – 18,0 % der Frachten des Kläranlagenzulaufes. Die vorliegende Arbeit enthält erstmals Ergebnisse zur Verfahrenskombination Überschussschlammdesintegration mit anschließender anaerob thermophiler Schlammstabilisierung. Gemäß den Versuchsauswertungen werden das anaerobe Abbauverhalten von Überschussschlämmen unter thermophilen Milieubedingungen und die daraus resultierenden Prozess-wasserbelastungen durch eine vorgeschaltete Überschussschlammdesintegration nur unwesentlich beeinflusst. Abschließend ordnet die Arbeit das Leistungsvermögen der Verfahrenskombination Über-schussschlammdesintegration + anaerob mesophile Schlammstabilisierung gegenüber anderen Optimierungsmöglichkeiten der anaeroben Schlammstabilisierung ein. Die Untersuchungen zeigen, dass durch einen Wechsel des Temperaturniveaus der anaeroben Schlammstabilisierung vom mesophilen in den thermophilen Bereich der unter mesophilen Bedingungen erreichbare GV Abbau bzw. zufuhrspezifische Biogasanfall unbehandelter Überschussschlämme ähnlich beeinflusst werden kann, wie es durch die Kombination von Überschussschlammdesintegration und anschließender anaerober mesophiler Schlammstabilisierung möglich ist. Gleichzeitig ist bei der anaerob thermophilen Stabilisierung unbehandelter Überschussschlämme gegenüber der anaerob mesophilen Stabilisierung desintegrierter Überschussschlämme eine höhere Prozesswasserbelastung zu erwarten.

Submitted in complete fulfillment for the degree of Doctor of Philosophy (Biotechnology), Durban University of Technology, Durban, South Africa, 2015. / Anaerobic digestion, a proven and highly efficient biological process for treating industrial wastewater and biogas generation is an underutilized technology in South Africa. Some of the industries that have on-site anaerobic reactors tend to face problems in operating these reactors due to poor understanding of the process and implementation of the technology. This has resulted in high pollutant loads in their final effluents and low energy recovery. In this study, an on-site full–scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was extensively monitored in order to evaluate the efficiency in terms of effluent quality, biogas production and microbial structure. Furthermore, developed and adopted kinetic models were used to optimize the performance of the full–scale UASB reactor using a combined Pareto differential evolution (CPMDE) algorithm. A preliminary analysis of the raw wastewater has shown that the wastewater produced from the brewery industry was high in organic matter with a total chemical oxygen demand (COD) between 1096.41 to 8926.08 mg/L. The average removal efficiency of COD from the UASB reactor after treatment was 79% with a methane (CH4) production of 60-69% at temperature ranges of 28-32˚C and hydraulic retention time (HRT) of 12 h within the optimal pH range for anaerobic bacteria (6.6 and 7.3) under various organic loading rates. However, the results also showed an increase in total suspended solids (TSS), nitrogen (N2), ammonia (NH3) and orthophosphate concentrations when comparing the influent to the effluent, which indicated the necessity for further optimization of the reactor condition in order to reduce these effluent parameters to acceptable standards and to increase CH4 production. In order to optimize the process, a thorough understanding of microbial interaction was essential. A combination of different molecular techniques viz., fluorescence in–situ hybridization (FISH), polymerase chain reaction (PCR) and quantitative real-time PCR (QPCR) were employed to understand the microbial community structure of the granular sludge samples using species specific primers and probes. The results revealed that the dominance of diverse groups of eubacteria belonging to phyla Proteobacteria, Firmicutes and Chloroflexi and an uncultured candidate division WS6 with four different orders of methanogenic Archaea viz., Methanomicrobiales, Methanococcales, Methanobacteriales and Methanosarcinales belonging to hydrogenotrophic and aceticlastic methanogens were within the reactor samples. Quantification of the 16S rDNA copies of eubacteria and methanogenic Archaea using species-specific primers further confirmed the spatial distribution of these microorganisms within the different compartments of the reactor where, the upper compartments were dominated by eubacteria and the lower compartments by methanogenic Archaea. The concentration of Archaea per nanogram of DNA was much higher (96.28%) than eubacteria (3.78%) in lower compartments, while, the eubacteria concentration increased to 98.34% in upper compartments with a decrease in Archaea quantity (1.66%). A modified kinetic methane generation model (MMGM) was developed on the basis of mass balance principles with respect to substrate (COD) degradation and the endogenous decay rate to predict CH4 production efficiency of the reactor. Furthermore, a Stover–Kincannon kinetic model was adopted with the aim of predicting the final effluent quality in terms of COD concentration and model coefficients were determined using the data collected from the full–scale reactor. Thereafter, a model-based multi-objective optimization was carried out using the CPMDE algorithm with three–objective functions namely; maximization of volumetric CH4 production rate; minimization of effluent substrate concentration and minimization of biomass washout, in order to increase the overall efficiency of the UASB reactor. Important decision variables and constraints related to the process were set for the optimization. A set of non-dominated solutions with high CH4 production rates of between 2.78 and 5.06 L CH4/g COD/day at low biomass washout concentrations were obtained at almost constant solution for the effluent COD concentration. A high COD removal efficiency (85-87%) at ~30-31˚C and 8-9 h HRT was obtained for the multi-objective optimization problem formulated. This study could significantly contribute towards optimization of a full–scale UASB reactor treating brewery wastewater for better effluent quality and biogas production. Knowledge on the activity and performance of microbial community present in the granular sludge taken from the full–scale UASB reactor would contribute significantly to future optimization strategies of the reactor. In addition, optimization using an evolutionary algorithm under different operational conditions would help to save both time and resources wasted in operating anaerobic bioreactors.