Microbial adaptation and response to high ammonia concentrations and precipitates during anaerobic digestion under psychrophilic and mesophilic conditions

Abstract

This study explored microbial adaptation to high ammonia concentrations (<1000 mg/L to 4000 mg/L) during anaerobic digestion (AD) under psychrophilic and mesophilic conditions, the latter of which yielded precipitates facilitating investigation of microbial response. The experimental setup was performed at bench-scale using microbial consortia from four different operating anaerobic digesters treating different organic wastes (WW-wastewater sludge, MN-manure, FW- food waste and CO-co-digestion (FW & MN)). Adaptation experiments were conducted with semi-continuous flow mode to resemble large-scale operation. Metagenome and 16S RNA analysis were performed for the first time in a psychrophilic reactor during an ammonia acclimation process. These analyses were also performed in mesophilic reactor exposed to precipitates and high ammonia levels. Diversity reduced when adaptation occurred successfully from 1.1 to 4 g/L of total ammonia nitrogen (TAN) under psychrophilic conditions, while the microbial community became more diverse under mesophilic conditions with ammonia inhibition. We report for the first time Methanocorposculum as a robust hydrogenotrophic methanogen at high ammoniacal concentrations under psychrophilic conditions. Additionally, Methanosarcina was present in low and high ammoniacal concentrations in mesophilic conditions, but there was a shift in species dominance. Methanosarcina barkeri stands out as a more resilient methanogen compared to Methanosarcina mazei, which initially dominated at <1.1 g/L TAN. We also explored the effects of sudden precipitates on methanogenic communities and methane production when they occurred under mesophilic conditions in two reactors. Methane production declined by more than 50% when precipitates occurred and was accompanied by pH reduction and VFA accumulation. Diversity data corroborated that methanogens were severely reduced. These two reactors were not able to recover with 50 days of added operation, demonstrating potential for long-term negative impacts of precipitate formation on AD performance stemming from negative impact to methanogenic communities.

Keywords: Ammonia adaptation; Metagenomics; Microbial function; Microbial structure; Psychrophilic reactor; Struvite precipitation.