Psychrophilic (8 °C) anaerobic treatment of partly acidified waste water was investigated using a two stage EGSB system with a total volume of 8.6 dm3. The reactor system was operated at an up-flow velocity of 10 m·h−1 and was fed with a sucrose-VFA mixture of 550–1100 mg COD dm−3. The average CODsol and VFA-COD removal efficiencies were 97 and 90 %, respectively, at total organic loading rates (OLR) ranging between 5.1–6.7 g COD dm−3·day−1, sucrose loading rates up to 1 g COD dm−3 day−1 and a hydraulic retention time (HRT) of 4 h. An increase in the sucrose loading rates resulted in a significant wash-out of biomass from the first stage. The second stage satisfactory served as a scavenger of non-degraded VFA from the first stage.
Specific activity assays showed an increase of 15 % in the specific methanogenic activity of the sludge present in the second stage and a decrease of 9 % in the first stage. Apparently, an enrichment of methanogens and acetogens in the anaerobic sludge in the second stage took place at temperatures as low as 8°C. The acidogenic population became much more dominant in the first stage, resulting in a higher acidifying activity and a decreased methanogenic activity. 16S rRNA probe-techniques (dot blot hybridization) showed that the acetate consuming Methanosaeta (formerly Methanothrix) and the hydrogenotrophic Methanobrevibacter species (or relatives) were the most abundant methanogens present in the psychrophilic sludge. The ratio between bacterial and methanobacterial hybridization signal of the first stage was 3 times higher than that of the second stage. By using NMR techniques, a higher effective diffusion coefficient was found for the smaller sized granules in both reactors, which is in congruent with the higher maximum specific acetate degrading activity of the smaller granules.