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In order to enhance the efficiency of anaerobic digestion, the effects of ultrasounds, ozonation and thermal pre-treatment have been studied on waste activated sludge. The feature of this study was to carry out the comparison of the three pre-treatments in the same conditions and on the same sludge sample. Each treatment was tested in two conditions close to optimum conditions to maximise batch anaerobic sludge biodegradability. All treatments led to chemical oxygen demand and matter solubilisation and had little influence on mineral matter. In terms of solubilisation thermal pre-treatment was better than sonication or ozonation. But, in terms of batch anaerobic biodegradability, best results were obtained with ultrasounds with an energy of 6250 or 9350 kJ/kg TS and a thermal treatment at 170 or 190 • C. Moreover, treatments had effects on physicochemical characteristics of sludge samples: apparent viscosity decreased after all treatments but the reduction was more important with thermal treatment. Median diameter of sludge flocs were reduced after sonication, increased after thermal treatment and did not change after ozonation. Finally, capillary suction time (CST) increased after ozonation, increased highly after sonication and was reduced after thermal treatment.

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The diversity of Archaea in anaerobic digesters was characterized by strand conformation polymorphism (SSCP) analysis and the sequencing of 16S rDNA genes. The 44 digesters sampled, located in eight different countries, treated effluents from agriculture, the food processing and petro-chemical industries, pulp and paper plant, breweries, slaughterhouses and municipal waste. All the existing processes were represented among the samples (fixed-film, fluidized bed, stirred-tank, UASB, sequential batch reactor, lagoon). Single strand conformation polymorphism analysis targeting the V3 region of 16S rDNA revealed between four to six distinct archaeal peaks per digester. The diversity of dominant Archaea in the 44 digesters was estimated as 23 different 16S rDNA sequences. Cloning of archaeal 16S rRNA genes from 11 distinct total genomic DNA, screening of clones by SSCP and the sequencing of 170 of them made it possible to characterize these SSCP peaks. All the sequences retrieved were members of the Euryarchaeaota subdomain. Furthermore, most of the sequences retrieved were very close to already known and cultivated strains or to environmental clones. The most frequent archaeal sequences were close to Methanosaeta concilii and to a 16S rDNA clone vadinDC06 located in the Methanobacterium clade (84% and 73% of digesters respectively). The other sequences were members of the Methanobacteriales and the Methanomicrobiales families. Only one sequence was far from any sequence of the database and it could be grouped with several sequences of environmental clones. Each digester harboured between two to nine archaeal sequences with only one of them corresponding to a putative acetate-utilizing species. Furthermore, the process in the digesters appeared to play a part in the distribution of archaeal diversity.