The unscientific disposal of agricultural solid waste introduces more antibiotics and other pollutants into the environment. Composting, as an environmentally friendly solid waste disposal method, can be used as a green way to degrade antibiotics, and conditioners can regulate the physicochemical indicators of the composting process. This article investigates the removal mechanism of tetracycline antibiotics (TCs) during the composting process by adding different regulators (biochar, zeolite, and biochar + zeolite). The results showed that the conditioning agent could significantly improve the removal efficiency and removal rate of TCs in compost. Among them, the addition of the zeolite group had the highest degradation rate of TCs, which were 91.39% (Tetracycline), 97.18% (Chlortetracycline), and 95.68% (Oxytetracycline). The combination of biochar and zeolite conditioning agents effectively minimized the migration of TCs into the soil. According to the findings of the artificial neural network model, it was determined that TCs exhibited the highest sensitivity to biochar + zeolite modulators at 31.28%. Conditioners influenced the removal of TCs in compost by impacting their physicochemical properties and microbial community structure. We isolated and domesticated a suitable microbial preparation that promotes the degradation of TCs, including Acinetobacter pittii, Stenotrophomonas maltophilia, Lactobacillus reuteri, Pseudomonas putida, and Trichosporon dohaense.