Polystyrene nanoplastics reshape the anaerobic granular sludge for recovering methane from wastewater

Wastewater has been identified as an important carrier for nanoplastics, which could elicit unintended impacts on critical microbial processes. However, the long-term impacts of nanoplastics on anaerobic granular sludge (AGS) for methane recovery from wastewater and the mechanisms involved remains unclear. In this study, we investigated the long term exposure-response relationship between polystyrene nanoplastics (Nano-PS) and AGS. In continuous test over 120 days with 86 days' Nano-PS exposure, feeding wastewater with 10 μg/L of Nano-PS had no significant impacts on the AGS performance. In comparison, higher levels (i.e., 20 and 50 μg/L) of Nano-PS decreased methane production and chemical oxygen demand (COD) removal by 19.0-28.6% and 19.3-30.0%, respectively, along with volatile fatty acids (VFA) accumulation. More extracellular polymeric substance (EPS) was induced by 10 μg/L of Nano-PS as a response to protect microbes, but higher levels (i.e., 20 and 50 μg/L) of Nano-PS decreased EPS generation, causing a decline in granule size and cell viability. Fluorescence tagging found that a large number of Nano-PS agglomerated/accumulated on the outer layer of AGS and even transferred into deeper layers of AGS over exposure time, producing toxic effects to adherent microorganisms, e.g., Longilinea sp., Paludibacter sp. and Methanosaeta sp.. The oxidative stress induced by Nano-PS was revealed to be a key factor for reshaping the AGS, reflected by the increased reactive oxygen species (ROS) generation and lactate dehydrogenase (LDH) release. The sodium dodecyl sulfate (SDS) leached from Nano-PS was also demonstrated to restrain the activities of antioxidant enzymes, thereby further lessening resistance to oxidative stress induced by Nano-PS. This work improves our ability to predict the risks associated with this ubiquitous contaminant in the environment.