Enhancing production of methane from cellulose is of utmost importance to improve the fermentation efficiency of lignocellulosic biomass. Antibiotics have demonstrated their ability to stimulate anaerobic digestion (AD) by influencing micro-organism activity. However, there has been limited research on the specific effect of antibiotics on cellulose AD. In this study, we investigated the impact of three antibiotics—tetracycline (TC), cephalexin (CLX), and azithromycin (AZM)—on cellulose AD when inoculated with waste sewage sludge. The results revealed that the presence of AZM and TC led to significantly higher methane yields, with increases of 51.94% and 34.96%, respectively, during a 20-day AD period. In contrast, the presence of CLX resulted in a 23.95% lower methane yield compared to the control. Furthermore, detailed analyses indicated that AZM had a positive influence on cellulose AD at all stages, including methanogenesis, acidogenesis/acetogenesis, and hydrolysis. On the other hand, TC primarily promoted AD during the methanogenesis stage. These findings collectively offer valuable guidance for efficiently transforming the energy potential of lignocellulosic wastes.

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