Preservation techniques and carbon and nitrogen growth enhancement of batch cultivated UASB granules

Cameron, Michelle (2000-12)

Thesis (MSc Food Sc )--Stellenbosch University, 2000.

Thesis

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.

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