Selective culture media were utilized for mixed culture of photosynthetic bacterias(PSB)enrichment and cultivation,and the microorganisms were sampled from natural multiple sludge pools with plenty of sunlight and high organic contents.A cell partial reflux device was employed for the strain screening and optimization.Then optimum reflux parameters were determined,and the mixed PSB which had high activity of hydrogen production was obtained.The hydrogen production characteristics of mixed culture by using cattle manure wastewater were also investigated in this paper.The results showed that,the mixed cultivation of PSB was indicated that the mixed PSB culture grew fast and stable,and the conditions for its growth and hydrogen production were less strict than those of pure PSB.