Microbial community analysis during continuous fermentation of thermally hydrolysed waste activated sludge.

Acidogenic fermentation of thermally hydrolysed waste activated sludge was carried out at laboratory scale in two reactors operated under different hydraulic retention times (HRT). Process performance was assessed in terms of volatile fatty acid (VFA) composition and yield. The diversity of the microbial population was investigated by constructing a 16S rRNA gene library and subsequent phylogenetic analysis of clones. Fluorescence in situ hybridization (FISH) was used to assess the relative abundance of different bacterial groups. Bacteroidetes and Firmicutes were the dominant taxonomic groups representing 93% of the total sequences obtained in the reactor with 4 d HRT. A similar VFA yield (0.4-0.5 g VFA(COD) g SCOD(-1)) was obtained for the HRTs tested (1-4 d), indicating that extended retention times were not useful. Within Firmicutes, Clostridia was the major group detected in the clone sequences. These had close affiliation to Sporanaerobacter acetigenes, suggesting organisms of this group were important for hydrolysis of the protein fraction of the substrate. However, FISH analysis failed to detect the major portion of the bacteria, and this is most likely due to the lack of appropriate probes. This work emphasizes the diversity of fermentative communities, and indicates that more work is needed to identify and detect the important members.

Enhanced solubilisation of the residual fraction of municipal solid waste.

The Swedish ordinance banning the landfilling of organic material after 2005 has led to rapid developments towards waste incineration, while biological alternatives have been less studied. In this study, biological alternatives for enhanced methane production from residual municipal waste (the remaining waste fraction after source separation) are investigated. The strategies investigated were recirculation of leachate, recirculation of leachate with aeration, flooding of the solid bed, and enzyme addition after initial leaching in an anaerobic, batch, two-stage digestion process with recirculation of digested leachate. The degree of solubilisation of organic compounds achieved was higher for initial digestion in a two-stage anaerobic digestion system followed by the addition of cellulolytic enzymes than in the other strategies investigated. The overall net solubilisation achieved was 0.48 g COD/g VSadded corresponding to an increase of 34%. In addition, the digestion time was considerably reduced using this strategy. For the other strategies investigated the solubilisation yields obtained were similar, 0.31 g COD/g VSadded.

Hydrolysis and microbial community analyses in two-stage anaerobic digestion of energy crops.

AIMS: The roles of the diverse populations of micro-organisms responsible for biodegradation of organic matter to form methane and carbon dioxide are rudimentarily understood. To expand the knowledge on links between microbial communities and the rate limiting, hydrolytic stage of two-stage biogas production from energy crops, this study was performed. METHODS AND RESULTS: The process performance and microbial communities (as determined by fluorescence in situ hybridization) in two separate two-stage batch digestions of sugar beets and grass/clover were studied. The microbial populations developed in the hydrolytic stage of anaerobic digestion of beets and grass/clover showed very few similarities, despite that the hydrolysis dynamics were similar. In both substrates, the solubilization of organic material was rapid for the first 10 days and accompanied by a build-up of volatile fatty acids (VFAs) and lactate. Between days 10 and 15, VFA and lactate concentrations decreased, as did the solubilization rates. For both substrates, Archaea started to appear in the hydrolytic stage between days 10 and 15, and the fraction of Bacteria decreased. The major bacterial group detected in the leachate fraction for beets was Alphaproteobacteria, whereas for grass/clover it was Firmicutes. The number of cells that bound to probes specifically targeting bacteria with cellulolytic activity was higher in the digestion of grass than in the digestion of beet. CONCLUSIONS: This study allowed the identification of the general bacterial groups involved, and the identification of a clear shift in the microbial population when hydrolysis rate became limiting for each of the substrates investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings from this study could be considered as a first step towards the development of strategies to stimulate hydrolysis further and ultimately increasing the methane production rates and yields from reactor-based digestion of these substrates.