Chemical and bacteriological composition of granular methanogenic sludge

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Abstract
Methanogenic sludge granules grown on waste water from a sugar refinery consisted of several bacterial morphotypes embedded in a matrix of extracellular material. Comparison of critical point drying and freeze-drying methods for preparing samples for scanning electron microscopy to determine the presence of extracellular material indicated that the former method permitted observations of extracellular material and intact cells. The effects of different extraction methods used for isolation of these extracellular polymers was also investigated by scanning electron microscopy. Of the various extraction procedures (EDTA, NaOH, autoclaving, water–phenol), water–phenol left most of the cells intact and was found to be a very efficient method of extraction. Extracellular polymers equivalent to 10–20 mg hexose/g of granules were extracted. The high resistance of the granules against disintegration by various chemical methods suggested that different extracellular polymers and probably different groups of organisms contributed to the matrix in which the bacteria were embedded. The chemical composition of the granules did not differ from the composition of bacteria in general. The buoyant density of 1.00–1.05 g∙g−1of the granules indicated that a simple agglomeration was the mechanism by which these methanogenic consortia improved their settling characteristics.