With the burgeoning growth of aquaculture industry, high concentration of NH4+-N, phosphorus and tetracycline are the prevalent pollutants in aquaculture wastewater posing a significant health risk to aquatic organisms. Therefore, an effective method for treating aquaculture wastewater should be urgently explored. Simultaneous removal of NH4+-N, phosphorus, tetracycline, and chemical oxygen demand (COD) in aquaculture wastewater was developed by moving bed biofilm reactor (MBBR) under co-metabolic substances. The result showed that co-metabolism substances had different effects on MBBR performance, and 79.4 % of tetracycline, 68.2 % of NH4+-N, 61.3 % of total nitrogen, 88.3 % of COD, and 38.1 % of total phosphorus (TP) were synchronously removed with sodium acetate as a co-metabolic carbon source. Protein (PN), polysaccharide (PS), and electron transfer system activity were used to evaluate the MBBR performances, suggesting that PN/PS ratio was 1.48, 0.91, 1.07, 3.58, and 0.79 at phases I-V. Additionally, a mode of tetracycline degradation and TP removal was explored, and the cell apoptosis was evaluated by flow cytometry. The result suggested that 74 %, 83 %, and 83 % of tetracycline were degraded by extracellular extracts, intracellular extracts, and cell debris, and there was no difference between extracts and non-enzyme in TP removal. The ratio of viable and dead cells from biofilm reached 33.3 % and 7.68 % with sodium acetate as a co-metabolic carbon source. Furthermore, Proteobacteria and Bacteroidetes in biofilm were identified as the dominant phyla for tetracycline and nutrients removal. This study provides a new strategy for tetracycline and nutrients removal from aquaculture wastewater through co-metabolism.