The effect of tetracycline (TC) on nitrogen removal in wastewater treatment plants has become a new problem. This study investigated the effects of TC on nitrogen removal using a Moving Bed Biofilm Reactor system. The results showed that there was no significant effect on nitrogen removal performance when the concentration of TC was 5 mg/L, and that the total nitrogen (TN) removal efficiency could reach 75-77%. However, when the concentration of TC increased to 10 mg/L, the denitrification performance was affected and the TN removal efficiency decreased to 58%. The abundance of denitrifying bacteria such as those in the genus Thauera decreased, and TC-resistant bacteria gradually became dominant. At a TC concentration of 10 mg/L, there were also increases and decreases, respectively, in the abundance of resistance and denitrification functional genes. The inhibitory effect of TC on denitrification was achieved mainly by the inhibition of nitrite-reducing bacteria.