Size- and Dose-Dependent Modulation of Methane Production by Polyethylene Microplastics During Anaerobic Digestion of Waste Activated Sludge

Polyethylene microplastics (PE-MPs) are ubiquitous constituents of waste activated sludge (WAS), acting as a major land-based source threatening coastal environmental integrity. However, how particle size and dose govern the methanogenic outcome during WAS digestion remains poorly defined. This study evaluated two particle sizes (50 vs. 300 µm) and doses (100 vs. 200 particles/gTS) to elucidate the differential effects of PE-MPs on methane yield and the underlying biological mechanisms. The results show that, while low-dose treatments either slightly inhibited methane yield (RS1) or had no significant effect (RL1), high-dose treatments (RS2 and RL2) achieved a net positive effect, with significant increases of 10.2% (p < 0.05) and 9.0% (p < 0.05) relative to the control, respectively. Nevertheless, RS2 and RL2 achieved methanogenic enhancement via distinctly different biological pathways. RS2 harnessed the stress of reactive oxygen species (ROS) (110.5% of the control) to drive community restructuring and biomass accrual (positive correlation between ROS intensity and total VS, Pearson’s r = 0.99). Key syntrophic and electrogenic taxa (e.g., Syntrophales, Bacteroidetes vadinHA17) exhibited a fully interconnected, decentralized network, thereby achieving tight coupling between hydrolysis and methanogenesis. RL2 leveraged the physical carrier effect to promote granulation and biomass growth, enriching Syntrophobacter to enhance propionate degradation. This culminated in a highly modular, sparse network characterized by localized competitive interactions. Together, dosage governs the net methanogenic effect of PE MPs, whereas particle size dictates the mechanistic routes of action. This work offers a mechanistic framework to optimize energy recovery from PE-MP-contaminated sludge while mitigating secondary environmental risks, providing a science-based strategy for the sustainable management of plastic-laden sludge that reconciles renewable energy recovery with pollution control.