Abstract

This study aimed to improve anaerobic digestion (AD) efficiency through the addition of zero-valent iron (ZVI) and biogas slurry. This paper demonstrated that methane production was most effectively promoted at a biogas slurry reflux ratio of 60%. The introduction of ZVI into anaerobic systems does not enhance its bioavailability. However, both biogas slurry reflux and the combination of ZVI with biogas slurry reflux increase the relative abundance of microorganisms involved in the direct interspecific electron transfer (DIET) process. Among them, the dominant microorganisms Methanosaeta, Methanobacterium, Methanobrevibacter, and Methanolinea accounted for over 60% of the total methanogenic archaea. The Tax4Fun function prediction results indicate that biogas slurry reflux and the combination of ZVI with biogas slurry reflux can increase the content of key enzymes in the acetotrophic and hydrotrophic methanogenesis pathways, thereby strengthening these pathways. The corrosion of ZVI promotes hydrogen production, and the biogas slurry reflux provided additional alkaline and anaerobic microorganisms for the anaerobic system. Their synergistic effect promoted the growth of hydrotrophic methanogens and improved the activities of various enzymes in the hydrolysis and acidification phases, enhanced the system's buffer capacity, and prevented secondary environmental pollution. PRACTITIONER POINTS: Optimal methane production was achieved at a biogas slurry reflux ratio of 60%. Biogas slurry reflux in anaerobic digestion substantially reduced discharge. ZVI addition in combination with biogas slurry reflux facilitates the DIET process.

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