Tetracycline has been frequently detected in municipal wastewater due to its extended use for various purposes. This study investigated the influence of tetracycline on non-aerated microalgal-bacterial granular sludge cultivated for municipal wastewater treatment. It was found that ammonia-N removal rate decreased at the tetracycline concentrations of 1 and 10 mg/L. A mass balance on nitrogen further revealed that the observed ammonia-N removal could be mainly attributed to microalgal assimilation which was inhibited by tetracycline at the concentrations studied. In fact, reduced production of chlorophyll in microalgae was observed in the presence of tetracycline, leading to decreased ammonia-N removal rate. Meanwhile, decreased dissolved oxygen (DO) concentration at high tetracycline concentration also indicated inhibition of microalgae. Furthermore, the relative abundances of microalgae containing green algae and cyanobacteria were inhibited by tetracycline. The results gathered in this study indicated the tetracycline-induced decoupling of symbiosis in microalgal-bacterial granular sludge. It is expected that this study can shed lights on the behaviors of non-aerated microalgal-bacterial granules in response to the presence of tetracycline during municipal wastewater treatment.