This study presented an approach by combining the real-time reverse transcription polymerase chain reaction with the terminal restriction fragment length polymorphism (T-RFLP) to investigate transcriptional responses of ammonia-oxidizing bacteria (AOB) to dimethyl sulfide (DMS) inhibition. Batch experiments with added ammonium and DMS were conducted using three activated sludges and Nitrosomonas europaea, and the transcriptional responses of the amo subunit A (amoA) mRNA were evaluated. It was found that DMS inhibited ammonium oxidation and amoA mRNA expression in all batch experiments but the inhibition degree observed was different for different sludges examined. It is likely that the different inhibitory effects of DMS on ammonium oxidation and amoA mRNA expression stemmed from different dominant AOB populations in the sludges. The T-RFLP results for amoA mRNA suggested that inhibition of ammonium oxidation by DMS to Nm. europaea-like AOB with T-RF 219/270 is relatively minor compared to other AOB populations in the examined sludges, such as Nm. europaea-like AOB with T-RF 491/491. (C) 2014 Elsevier Ltd. All rights reserved.