Three laboratory-scale moving bed biofilm reactors (MBBR) with different carrier filling ratios ranging from 40% to 60% were used to study the effects of carrier-attached biofilm on oxygen transfer efficiency. In this study, we evaluated the performance of three MBBRs in degrading chemical oxygen demand and ammonia. The three reactors removed more than 95% of NH4+-N at an air flow-rate of 60 L·h^-1. The standard oxygen transfer efficiency (αSOTE) of the three reactors was also investigated at air flow-rates ranging from 60 to 100 L·h^-1. These results were compared to αSOTE of wastewater with a clean carrier (no biofilm attached). Results showed that under these process conditions, αSOTE decreased by approximately 70% as compared to αSOTE of wastewater at a different carrier-filling ratio. This indicated that the biofilm attached to the carrier had a negative effect on αSOTE. Mechanism analysis showed that the main inhibiting effects were related to biofilm flocculants and soluble microbial product (SMP). Biofilm flocs could decrease αSOTE by about 20%, and SMP could decrease αSOTE by 30%–50%.