Adhesion of bacteria involved in anaerobic consortia was investigated in upflow anaerobic sludge bed reactors and was related to surface thermodynamics. The adhesion of hydrophilic cells appeared to be enhanced at a low liquid surface tension (␥ LV), while the adhesion of hydrophobic cells was favored at a high ␥ LV. Growth in protein-rich growth media resulted in low granular biomass yields; addition of polycations, such as poly-L-lysine and chitosan, increased the ␥ LV and the granular biomass yield. On the basis of the results of activity tests and microbial counts with wash-out cells, we identified two types of structured granules that were related to the influence of ␥ LV. In one type of granules, hydrophilic acidogens surrounded a more hydrophobic methanogenic association. These granules were selected at a low ␥ LV provided that carbohydrates were available as substrates. The other type of granules was selected at a high ␥ LV ; hydrophobic cells (i.e., methanogens) were predominant throughout these granules. The granules which had acidogens as solid-phase emulsifiers around a methanogenic association appeared to allow more stable reactor performance. Decreasing the ␥ LV in the reactor by adding trace amounts of a surfactant also increased reactor stability.

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The influence of a high energy substrate, i.e. sucrose, on the granular sludge yield and the development of different types of granular sludge was investigated by using Upflow Anaerobic Sludge Bed (UASB) reactors fed with synthetic wastewater. The feed COD was a mixture of volatile fatty acids (VFA) i.e., 20, 40, and 40% of the COD as C2-, C3-, and C4-VFA, respectively. Furthermore, experiments were carried out in which 10 and 30% of the VFA COD was substituted with sucrose. The following distinctly different types of granules were observed in each testrun: in the reactor fed with solely VFA, black (B) and white (W) granules developed; in the reactor fed with a mixture of 90% VFA and 10% sucrose, three types of granules i.e., B, W, and grey (G) granules could be seen; in the reactor fed with 70% VFA and 30% sucrose, only W and G granules were found. The granular sludge yield increased proportional to the amount of sucrose COD. At steady-state performance of the reactors, specific acidogenic (SAA) and methanogenic (SMA) activity tests on these granules revealed that B granules had the highest SMA with low SAA. The W granules had very high SMA with low SAA. G granules gave the highest SAA with a considerable SMA. Measurement of coenzyme F420 revealed that B granules consist mainly of acetoclastic methanogens. The fore-mentioned tests were supplemented with analyses of the wash-out cells present in the reactor effluent and the results suggested that acidogens, if present, prevail at the granule surface. The B granules were particularly rich in Ca, Mn, and Zn minerals. The size distribution analysis showed that the granule diameter increased in the following order: B < W < G granules. The biogas bubbles tended to adhere to the B and W granules but not so strongly to the G granules.