An in-vessel composting bioremediation of 13C4-4,5,9,10-pyrene and unlabeled pyrene spiked soil amended with fresh wastes was investigated by DNA-based stable isotope probing (SIP) of active bacteria involved. Highest dissipation of 13C4-pyrene was detected at 55 °C after 42 days composting. The active bacterial communities in the composting changed over time, showing a distinct difference among different stages. a-, b-, c-Proteobacteria, and Actinobacteria were detected mainly involving in pyrene degradation at 38 °C over 14 days composting. Streptomyces appeared to dominate the pyrene degradation at 55 °C. b- and c-Proteobacteria and Actinobacteria were the dominant pyrene degraders at 70 °C after 42 days composting and at 38 °C after 60 days composting. The results of this study suggest the pyrene degradation was performed by phylogenetically distinct bacterial guilds from the phyla Actinobacteria and Proteobacteria during in-vessel composition processes.