Due to the chemical and physical structure of a lignocellulosic biomass, its anaerobic digestion (AD) is a slow and difficult process. In this paper, the results obtained from a batch biochemical methane potential (BMP) test and fed-batch mesophilic AD assays of sunflower oil cake (SuOC) are presented. Taking into account the low digestibility shown during one-stage experiments the methane yield decreased considerably after increasing the organic loading rate (OLR) from 2 to 3 g VS L−1 d−1, SuOC was subjected to a two-stage AD process (hydrolytic-acidogenic and methanogenic stages), in two separate reactors operating in series where the methanogenic stage became acidified (with >1,600 mg acetic acid L−1) at an OLR as low as 2 g VS L−1 d−1. More recently, BMP assays were carried out after mechanical, thermal, and ultrasonic pre-treatments to determine the best option on the basis of the methane yield obtained.

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