Coffee mucilage (CM), a novel substrate produced as waste from agricultural activity in Colombia, the largest fourth coffee producer in the world, was used for hydrogen production. The study evaluated three ratios (C1-3) for co-digestion of CM and swine manure (SM), and an increase in organic load to improve hydrogen production (C4). The hydrogen production was improved by a C/N ratio of 53.4 used in C2 and C4. The average hydrogen production rate in C4 was 7.6 NL H-2/L(CM)d, which indicates a high hydrogen potential compare to substrates such as POME and wheat starch. In this condition, the biogas composition was 0.1%, 50.6% and 39.0% of methane, carbon dioxide and hydrogen, respectively. The butyric and acetic fermentation pathways were the main routes identified during hydrogen production which kept a Bu/Ac ratio at around 1.0. A direct relationship between coffee mucilage, biogas and cumulative hydrogen volume was established. (C) 2014 Elsevier Ltd. All rights reserved.