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.

Anaerobic process of biohydrogen production was developed recently. The representative strain of hydrogen production bacteria, namely fermentative hydrogen producing bacterial strain B49，was isolated from the activated sludge which was cultured artificially by our lab. In this study, there were some different C/N ratios employed. Experiment results indicated that the concentration of carbon and nitrogen had a great effect upon hydrogen production of B49, when C/N=2.5, glucose got the highest degraded efficiency, and at a C/N ratio of 3.3, the bio-gas and hydrogen production peaked with 159mL and 37.5mL, respectively, which considered that the optimum C/N ratio on the growth and hydrogen production was 3:1. Furthermore, the mainly terminal liquid products of B49 were ethanol and acetic acid. With the increase of the total organic nitrogen, final pH value decreased gradually.