Cellulose hydrolysis is the rate-limiting step in anaerobic digestion. In the present study, waste paper was used as a model of cellulosic biomass and was pretreated with rumen fluid prior to methane production. To achieve a high methane yield, the reaction time of the pretreatment was examined. Waste paper was soaked with rumen fluid for 6 and 24 h at 37 degrees C. Various volatile fatty acids, especially acetate, were produced by the pretreatment. Semicontinuous methane production was carried out over a 20-day period. The best daily methane yield was obtained by the 6-h pretreatment. The amount was 2.6 times higher than that of untreated paper, which resulted in 73.4% of the theoretical methane yield. During methane production, the cellulose, hemicellulose and lignin degradabilities were improved by the pretreatment. Pretreatment by rumen fluid is therefore a powerful method to accelerate the methane yield from a cellulosic biomass. (C) 2012 Elsevier Ltd. All rights reserved.