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.