An in-depth analysis of microbial response to exposure to high concentrations of microplastics in anaerobic wastewater fermentation.

The impact of microplastics (MPs) in anaerobic wastewater treatment on microbial metabolism is significant. Anaerobic granular sludge (AS) and biofilm (BF) are two common ways, and their responses to microplastics will have a direct impact on their application potential. This study investigated the microbial reactions of AS and BF to three types of MPs: polyethylene (PE), polyvinyl chloride (PVC), and a mixture of both (MIX). Results exhibited that MPs reduced methane output by 44.65 %, 55.89 %, and 53.18 %, elevated short-chain fatty acid (SCFA) levels by 95.93 %, 124.49 %, and 110.78 %, and lowered chemical oxygen demand (COD) removal by 28.77 %, 36.78 %, and 33.99 % for PE-MP, PVC-MP, and MIX-MP, respectively, with PVC-MP showing the greatest inhibition. Meanwhile, microplastics also facilitated the relative production of reactive oxygen species (ROS, 40.29 %-96.99 %), lactate dehydrogenase (LDH, 20.01 %-75.02 %), and adenosine triphosphate (ATP, 26.64 %-43.80 %), while reducing cytochrome c (cyt c, 23.60 %-49.02 %) and extracellular polymeric substances (EPS, 17.44 %-26.58 %). AS and BF displayed distinct enzymatic activities under MPs exposure. Correspondingly, 16S-rRNA sequencing indicated that AS was mainly involved in acetate generation by Firmicutes, while BF performed polysaccharide degradation by Bacteroidota. Metatranscriptomic analysis showed AS to be rich in acetogens (Bacillus, Syntrophobacter) and methanogens (Methanothrix, Methanobacterium), while BF contained more fermentation bacteria (Mesotoga, Lentimicrobium) and electroactive microorganisms (Clostridium, Desulfuromonas) under MIX-MP. Moreover, BF exhibited higher glycolysis gene expression, whereas AS was more active in methane metabolism, primarily through the acetoclastic methanogenic pathway's direct acetate conversion. This study provides new insights into understanding the microbial response produced by microplastics during anaerobic wastewater digestion.