The widespread distribution of polycyclic aromatic hydrocarbons (PAHs) in groundwater has become an important environmental issue. Knowledge of microbial community changes could aid in identification of particular microorganisms that are capable of degrading PAHs in contaminated aquifers. Therefore, 16S rRNA gene clone library analysis was used to identify the archaeal and bacterial communities in an aquifer sediment microcosm used for anaerobic anthracene degradation under methanogenic conditions. A remarkable shift of the archaeal community structure occurred after anaerobic anthracene degradation, but the types of the abundant bacterial phyla did not change. However, a decrease of both archaeal and bacterial diversity was observed. Bacterial genera Bacillus, Rhodococcus and Herbaspirillum might have links with anaerobic anthracene degradation, suggesting a role of microbial consortia. This work might add some new information for understanding the mechanism of PAH degradation under methanogenic conditions.