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The effect of continuous feeding on the stability of aerobic granular sludge (AGS) was investigated. A lab-scale sequencing batch reactor (SBR) seeded with mature AGS was operated for 30 days under a continuous feeding pattern. Under these conditions, part of the AGS disintegrated, and much tree-like sludge and some filamentous bacteria grew on the surface of the remaining AGS. During the first 13 days, the sludge volume index (SVI) increases rapidly, and then becomes stable. Quantitative investigation of the extracellular polymeric substances (EPS) indicates that extracellular polysaccharides decrease while extracellular proteins first increase and then decrease. After about 15 days, the content of EPS becomes steady. EPS staining shows that the proteins contribute to the formation of the core of the AGS, whereas the α-glucopyranosyl polysaccharides and β-D-glucopyranose polysaccharides accumulate in the outer layers of the AGS. After 30 days, the positions of proteins and α-glucopyranosyl polysaccharides change slightly, but the number of β-D-glucopyranose polysaccharides accumulated at the outer layer increases greatly. EPS staining also indicates that the stability of AGS is dependent on the protein core, and β-D-glucopyranose polysaccharides could be used to evaluate whether the AGS is stable or not. The results of this study show that continuous feeding is not favorable for the stability of AGS because of the low substrate gradient.
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