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Anaerobic digestion and fermentation processes in wastewater sludge treatment are limited by several factors, including the slow breakdown of complex organic matter and solubilization of solids. In this study, thermochemical pretreatment of thickened waste activated sludge using high temperature (>170 °C) was investigated to understand the impact of the pretreatment on the volatile fatty acids (VFA) production and its fractions during the fermentation process. Furthermore, the influence the thermochemical pretreatment on sludge disintegration and methane recovery was investigated. A range of acidic and alkaline conditions over the pH range of 4.5-10 was examined. Sludge (pH adjusted) was exposed to hydrothermal pretreatment (HTP) at a temperature of 170 °C for 30 min. Pretreated samples were then subjected to batch fermentation and methane potential tests which revealed that acidic and alkaline conditions resulted in increased sludge solubilization during HTP. Acidic conditions were associated with a higher VFA production yield of up to 185 mg chemical oxygen demand/g total chemical oxygen demand. Alkaline conditions led to a higher methane production yield where the maximum yield (276 mL CH 4 /g total chemical oxygen demand added ) was found to occur at pH 10. Therefore, alkaline sludge used for fermentation has shown technical and economic feasibility for sludge carbon recovery.

The effect of limiting phosphorus (P) in activated sludge was investigated in laboratory-scale sequencing batch reactors (SBRs). Correlative microscopy revealed that P-limitation (COD:N:P = 100:5:0.05) leads to morphological changes in floc structure and the composition of extracellular polymeric substances (EPS). This was found to be accompanied by expression of quorum-sensing in an acyl homoserine lactone bioassay. Differential gene expression in relation to P-limitation was examined in a global profile using the Affymetrix Escherichia coli antisense genomic microarray. Three separate experiments were conducted where the impact of P-limitation was examined under batch conditions and in SBRs at stable operating conditions and within 3-7 days following a down-shift in P. Significant changes in open reading frames (ORF) and intergenic regions based on the E. coli microarray were observed. Several genes associated with cell structure, including slt, wbbH, fimH, amB, rfaJ and slp were found to be expressed. Quorum regulated genes were also found to be expressed including psiF which is known to be induced by P-starvation (92% confidence level; 1.45 log ratio).

Correlative microscopy was applied to study the influence of solids retention time on activated sludge floc structure. Conventional optical microscopy revealed flocs at lower SRTs (4 and 9 days) to be irregular in shape while flocs at higher SRTs (16 and 20 days) had a more spherical and compact structure. Flocs were examined by environmental scanning electron microscopy (ESEM) and by transmission electron microscopy (TEM) coupled with energy-dispersive spectroscopy (EDS). Distinctive differences in floc structure and the arrangement of EPS were revealed. Flocs from higher SRTs were less hydrated and were found to possess a dense EPS layer that covers much of the surface. Extracellular osmiophilic granules present in these flocs indicate that the cells at the higher SRT may produce more lipid-like material. This EPS layer appears to decrease the floc surface roughness and protects the interior cells from disruption by changes in the external environment. Sludge flocs at higher SRTs were found to be physically more stable than those at lower SRTs.