The objective of the study was to identify the optimum time interval for effluent removal after temporarily stopping stirring in otherwise continuously stirred tank reactors. Influence of temperature (10 and 55 degrees C) and active biogas process on passive separation of digested manure, where no outside mechanical or chemical action was used, within the reactor was studied in three vertical settling columns (100 cm deep). Variations in solids and microbial distribution at top, middle and bottom layers of column were assessed over a 15 day settling period. Results showed that best solids separation was achieved when digested manure was allowed to settle at 55 degrees C with active biogas process (pre-incubated at 55 degrees C) compared to separation at 55 degrees C without active biogas process (autoclaved at 120 degrees C, for 20 min) or at 10 degrees C with active biogas process. Maximum solids separation was noticed 24 h after settling in column incubated at 55 degrees C, with active biogas process. Microbiological analyses revealed that proportion of Archaea and Bacteria, absent in the autoclaved material, varied with incubation temperature, time and sampling depth. Short rod shaped bacteria dominated at 55 degrees C, while long rod shaped bacteria dominated at 10 degrees C. Methanosarcinaceae were seen more abundant in the surface layer at 55 degrees C while it was seen more common in the top and bottom layers at 10 degrees C. Thus, passive separation of digester contents within the reactor can be used effectively as an operating strategy to optimize biogas production by increasing the solids and biomass retention times. A minimum of 1-2 h "non-stirring" period appears to be optimal time before effluent removal in plants where extraction is batch-wise 2-4 times a day. (C) 2007 Elsevier Ltd. All rights reserved.