The efficiency of bioaugmentation strategy for enhancing biohydrogenesis at elevated organic load was successfully evaluated by augmenting native acidogenic microflora with three acidogenic bacterial isolates viz., Bacillus subtilis, Pseudomonas stutzeri and Lysinibacillus fusiformis related to phyla Firmicutes and Proteobacteria separately. Hydrogen production ceased at 50 g COD/l operation due to feed-back inhibition. B. subtilis augmented system showed higher H-2 production followed by L. fusiformis, P. stutzeri and control operations, indicating the efficacy of Firmicutes as bioaugmentation biocatalyst. Higher VFA production with acetic acid as a major fraction was specifically observed with B. subtilis augmented system. Shift in metabolic pathway towards acidogenesis favoured higher H-2 production. FISH analysis confirmed survivability and persistence of augmented strains apart from improvement in process performance. Bio-electrochemical analysis depicted specific changes in the metabolic activity after augmentation which also facilitated enhanced electron transfer. P. stutzeri augmented system documented relatively higher COD removal. (C) 2014 Elsevier Ltd. All rights reserved.