The influence of hydraulic retention time (HRT) on the microbial communities was evaluated in an anaerobic sequencing batch reactor (AnSBR) using organic waste from a restaurant as the substrate. The relationship among Lactobacillus, Clostridium and Bacillus as key micro-organisms on hydrogen production from organic solid waste was studied. The effect of the HRT (8-48 h) on the hydrogen production and the microbial community was evaluated. Quantitative PCR was applied to determine the abundance of bacteria (in particular, Enterobacter, Clostridium and Lactobacillus genera). An AnSBR fermentative reactor was operated for 111 cycles, with carbohydrate and organic matter removal efficiencies of 80 +/- 15 center dot 42% and 22 center dot 1 +/- 4 center dot 49% respectively. The highest percentage of hydrogen in the biogas (23 center dot 2 +/- 11 center dot 1 %), and the specific production rate (0 center dot 42 +/- 0 center dot 16 mmol H-2 gVS(added)(-1) d(-1)) were obtained at an HRT of 48 h. The decrease in the HRT generated an increase in the hydrogen production rate but decreasing the content of the hydrogen in the gas. HRT significantly influence the abundance of Enterobacter, Clostridium and Lactobacillus during the hydrogen production from food waste leading the hydrogen production as well as the metabolic pathways. The microbial analysis revealed a direct relationship between the HRT and the presence of fermentative bacteria (Enterobacter, Clostridium and Lactobacillus genera). Clostridium sp. predominated at an HRT of 48 h, while Enterobacter and Lactobacillus predominated at HRTs between 8 and 24 h.