Biohydrogenbacterium R3 sp.nov, as a typical strains of hydrogen production bacteria, was isolated from the activated sludge in the biohydrogen production reaction by using culture media of HPB-LR which was designed particularly for isolating anaerobic fermentation bacteria and an improved Hungate rolling tubes technique and a plate method of culture bottle (PMCB) which were used for counting and isolating the anaerobes by our lab. Two representative strains of hydrogen production bacteria, namely RL20, RL37, and a strain of no hydrogen production bacterium, namely RL16 were used as contrast analysis. Experiment results indicated that the distributed proportion of ethanol and acetic acid was 95%-99% in the total amount of VAFs (Volatile Fatty Acid) of R3 which were typical ethanol-type fermentation, and the total amount of ethanol and acetic acid was commensurate with that of propionic acid.