The bacterial numeration and microbial observation were made on granular sludge from laboratory single and two-phase UASB reactors. It was shown that the fermentative bacteria (group I), H2-producing acetogenic bacteria (group II) and methanogenic bacteria (group III) of granular sludge in single UASB reactor were 9.3 × 108−4.3 × 109, 4.3 × 107−4.3 × 108, 2.0−4.3 × 108, respectively, during the granulation process. The sludge of methanogenic reactor exhibited the similar results. That indicates there is no big difference between suspended and granular sludge, and bacterial population for three groups of anaerobic bacteria are similar. The formation of granular sludge depends mainly on the organization and arrangement of bacteria. An observation of granular sludge using electron microscope revealed that the fermentative bacteria and hydrogenotrophic methanogens existed on outer surface of granules, and aceticlastic methanogens and H2-producing acetogenic bacteria occupied the inner layer. A new syntrophic association between Methanosaeta sp. and Syntrophomonas sp. (even plus Methanobrevibacter sp.) was observed. Because Methanosaeta can effectively convert the acetate (the end product of propionate and butyrate) to methane, such a new syntrophic association is supposed to support the degradation of short fatty acids and high methanogenic activity of granular sludge. Based on structural pattern, a hypothesis on mechanism of granulation called “crystallized nuclei formation” is proposed.

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