Thermal hydrolysis (TH) treatment has been verified to effectively reduce antibiotic resistance genes (ARGs) in waste activated sludge. This study focused on the effect of TH parameters on ARGs reduction, and the optimal conditions basing on tetracycline (tet) resistance genes reduction rate in sludge phase were pH 3, temperature 160 degrees C and reaction time 2 h. The pH was found to play a critical role in tet genes reduction behavior. Additionally, the tet genes distributions in TH supernatant are considered as well because genes will release from solid to liquid phase as sludge cell breaking. The lowest genes content in TH supernatant was also observed at pH 3. Further investigation revealed that contents of DNA and organics released from sludge phase to liquid phase under pH 3 were less than that in neutral and alkaline condition; moreover, the organics in TH supernatant was found to protect DNA. Therefore, under the acidic condition, the genes had less releasing from the sludge phase and the DNA protection effect of organics was weaker, resulting in the lowest tet genes content in TH supernatant. Moreover, bacterial community structure after TH was closely related with the ARGs content. The bacterial on phylum and genus level showed various responses to TH pH values. Although the Firmicutes phylum and Escherichia-Shigella genus exhibited the stronger resistance and had higher accumulation in TH acidic condition, more possible total tet genes hosts were destroyed in acidic condition, causing less ARGs remaining in the sludge phase under pH 3. (c) 2021 Elsevier Ltd. All rights reserved.