Role of Methanosarcina in mercuric mercury transportation and methylation in sulfate-driven anaerobic oxidation of methane with municipal wastewater sludge

Sulfate-driven anaerobic oxidation of methane (AOM) and anaerobic digestion (AD) with municipal wastewater sludge containing heavy metals may provide favorable conditions for the biogeochemical transformation of mercury (Hg) by methanogens and methanotrophs. However, it remains largely unclear what Hg-methylators functioned and what role Methanosarcina played in these processes. Here, we performed sulfate-driven AOM following AD with Hg-containing wastewater sludge and investigated the role of microbes, especially Methanosarcina, in the biogeochemical transformation of Hg based on 16S rRNA amplicon and metatranscriptomic sequencing. Results showed that methylmercury (MeHg) concentrations and MeHg/total Hg ratios increased significantly, implying mercuric Hg [Hg(II)] methylation predominated MeHg demethylation. Desulfovibrio, Desulfobulbus and Methanosarcina dominated and thus likely played important roles in Hg(II) methylation, while Methanosarcina dominated and functioned in methane metabolism. In the presence of sulfate, differentially-expressed genes (DEGs) related to Hg transporting ATPase increased significantly, indicating Methanosarcina absorbed a large amount of Hg(II) and likely further methylated it to MeHg. No Hg response DEGs were found in the absence of sulfate, further confirming sulfate played an essential role in Hg cycle. Overall, these results suggest that controlling sulfate levels and Methanosarcina abundances in municipal wastewater could potentially mitigate MeHg risks to humans.