The changes in microbial community structures during fermentative hydrogen production process were investigated by analyzing 16S rDNA gene sequences using gamma irradiated sludge as inoculum. The experimental results showed that the microbial community structure of untreated sludge was very rich in diversity. After gamma irradiation, lots of species were inhibited, and species with high survival rates under radiation conditions became dominant. After fermentation, Clostridium butyrium and a sequence closely related to Clostridium perfringens ATCC 13124(T) (CP000246) became predominant, which were all common hydrogen producers. Microbial distribution analysis indicated that gamma irradiation was a good pretreatment method for enriching hydrogen-producing strains from digested sludge. (C) 2015 Elsevier Ltd. All rights reserved.