Zinc pyrithione induced volatile fatty acids promotion derived from sludge anaerobic digestion: Interrelating the affected steps with microbial metabolic regulation and adaptive responses.

The massive use of zinc pyrithione (ZPT, as broad-spectrum bactericides) resulted in its high levels in waste activated sludge (WAS) and affected subsequent WAS treatment. This work revealed the effects of ZPT on the volatile fatty acids (VFAs) during WAS anaerobic digestion, in which VFAs yield was enhanced by approximately 6-9 folds (from 353 mg COD/L in control to 2526-3318 mg COD/L with low level of ZPT (20-50 mg/g TSS)). The ZPT occurred in WAS enabled the acceleration of solubilization, hydrolysis and acidification processes while inhibited the methanogenesis. Also, the low ZPT contributed to the enrichment of functional hydrolytic-acidifying microorganisms (e.g., Ottowia and Acinetobacter) but caused the reduction of methanogens (e.g., Methanomassiliicoccus and Methanothrix). Meta-transcriptomic analysis demonstrated that the critical genes relevant to extracellular hydrolysis (i.e. CLPP and ZapA), membrane transport (i.e. gltI, and gltL), substrates metabolisms (i.e. fadj, and acd), and VFAs biosynthesis (i.e. porB and porD) were all upregulated by 25.1-701.3% with low level of ZPT. Specifically, the ZPT stimulus on amino acids metabolism for VFAs transformation was prominent over carbohydrates. Moreover, the functional species enabled to regulate the genes in QS and TCS systems to maintain favorable cell chemotaxis to adapt the ZPT stress. The cationic antimicrobial peptide resistance pathway was upregulated to blunt ZPT with the secretion of more lipopolysaccharide and activate proton pumps to maintain ions homeostasis to antagonize the ZPT toxicity for high microbial activities, the abundance of related genes was up-regulated by 60.5 to 524.5%. This work enlightened environmental behaviors of emerging pollutants on WAS anaerobic digestion process with interrelations of microbial metabolic regulation and adaptive responses.