Anaerobic bio-hydrogen production is the focus in the field of bio-energy resources. In this paper, a series of batch experiments were conducted to investigate the effects of several factors on anaerobic bio-hydrogen producing process carried out by pre-heated river sediments. The results showed that several factors such as substrate and its concentration, temperature and the initial pH value could affect the anaerobic bio-hydrogen production in different levels. At 35°C and the initial pH of 6.5, using glucose of 20,000mg COD/L as substrate, the highest hydrogen production of 323.8ml-H2/g TVS in a 100ml batch reactor was reached, the specific hydrogen production rate was 37.7ml-H2/g TVSh, and the hydrogen content was 51.2%. Thereafter using the same pre-heated river sediments as seed sludge, continuous anaerobic bio-hydrogen production was achieved successfully in a lab-scale CSTR with gas-separator. At the organic loading rate of 36kg COD/m3d, the highest hydrogen production was 6.3–6.7l-H2/l-reactord, the specific hydrogen production was 1.3–1.4mol-H2/mol-glucose, and the hydrogen content in the gas was 52.3%. The effluent of the bio-reactor contained some small molecular organics, mainly including ethanol, acetate, butyrate and their molar proportion is 1 : 1 : 0.6.

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