Abstract

Ammonia inhibition is an important reason for reactor failures and economic losses in anaerobic digestion. Its impact on acetic acid degradation is well-studied, while its effect on propionic and butyric acid degradation has received little attention and is consequently not considered in the Anaerobic Digestion Model No. 1 (ADM1). To compare ammonia inhibition of the degradation of these three volatile fatty acids (VFAs), we fed a mixture of them as sole carbon source to three continuous stirred tank reactors (CSTRs) and increased ammonium bicarbonate concentrations in the influent from 52 to 277 mM. The use of this synthetic substrate allowed for the determination of degradation efficiencies for the individual acids. While butyric acid degradation was hardly affected by the increase of ammonia concentration, propionic acid degradation turned out to be even more inhibited than acetic acid degradation with degradation efficiencies dropping to 31 and 65% for propionic and acetic acid, respectively. The inhibited reactors acclimatized and approximated pre-disturbance degradation efficiencies toward the end of the experiment, which was accompanied by strong microbial community shifts, as observed by amplicon sequencing of 16S rRNA genes and terminal restriction fragment length polymorphism (T-RFLP) of mcrA genes. The acetoclastic methanogen Methanosaeta was completely replaced by Methanosarcina. The propionic acid degrading genus Syntrophobacter was replaced by yet unknown propionic acid degraders. The butyric acid degrading genus Syntrophomonas and hydrogenotrophic Methanomicrobiaceae were hardly affected. We hypothesized that the ammonia sensitivity of the initially dominating taxa Methanosaeta and Syntrophobacter led to a stronger inhibition of the acetic and propionic acid degradation compared to butyric acid degradation and hydrogenotrophic methanogenesis, which were facilitated by the ammonia tolerant taxa Syntrophomonas and Methanomicrobiaceae. We implemented this hypothesis into a multi-taxa extension of ADM1, which was able to simulate the dynamics of both microbial community composition and VFA concentration in the experiment. It is thus plausible that the effect of ammonia on VFA degradation strongly depends on the ammonia sensitivity of the dominating taxa, for syntrophic propionate degraders as much as for acetoclastic methanogens.

Highlights

  • Biogas production is an important renewable energy source and organic waste treatment technology (Plugge, 2017)

  • Increasing the ammonia concentration in the influent to 277 mM in RNH3 and RNH3,HCl starting on day 21 was followed by an accumulation of volatile fatty acids (VFAs) and decline in methane production rates

  • Starting on day 73, after HCl was omitted from the medium of RNH3,HCl, the pH value in RNH3,HCl rose to 7.7 toward the end of the experiment, equal to RNH3, while total VFA concentrations remained at about 1 gCOD L−1

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Summary

Introduction

Biogas production is an important renewable energy source and organic waste treatment technology (Plugge, 2017). Several underlying and partly connected mechanisms of free ammonia inhibition after diffusion into a cell have been put forward and summarized by Krakat et al (2017): proton imbalance, change of the intracellular pH, increase in maintenance energy requirement, and inhibition of specific enzymatic reactions. These mechanisms have been solely discussed as explanations for ammonia inhibition in the context of methanogenesis. They might apply to other functional groups, for example proton-reducing bacteria degrading propionic or butyric acid

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