Anaerobic digestion (AD) and microbial electrochemical technologies (MET) can be combined in manifold ways. Recent studies show negative influences of AD effluents on the performance of pre-grown Geobacter spp.-dominated biofilm anodes. In this study, it was investigated how such biofilm anodes are affected by AD effluents. Therefore, experiments using AD effluents in different concentrations (0−100%) in combination with biofilms of different ages were performed. Furthermore, the activity of methanogens was inhibited and minimized by application of 2-bromoethanesulfonate (2-BES) and microfiltration, respectively. Biofilms pre-grown for 5 weeks show higher resistance against AD effluents compared to biofilms pre-grown for only 3 weeks. Nevertheless, adaptation of biofilms to AD effluents was not successful. Biofilm activity in terms of coulombic efficiency and maximum current density (j max ) dropped by factor 32.2 ± 3.2 and 38.9 ± 8.4, respectively. The application of 2-BES and microfiltration had positive effects on the biofilm activity. The results support the assumption that methanogens or further compounds not studied here, for example, protozoans, which may have been inhibited or removed by 2-BES application or microfiltration, have an immediate influence on the stability of Geobacter spp.-dominated biofilms and may limit their practical application in AD environments.

Apart from their archetypic use in anaerobic digestion (AD) methanogenic archaea are targeted for a wide range of applications. Using different methanogenic archaea for one specific application requires the optimization of culture media to enable the growth of different strains under identical environmental conditions, e.g., in microbial electrochemical technologies (MET) for (bio)electromethanation. Here we present a new culture medium (BFS01) adapted from the DSM-120 medium by omitting resazurin, yeast extract, casitone, and using a low salt concentration, that was optimized for Methanosarcina barkeri, Methanobacterium formicicum, and Methanothrix soehngenii. The aim was to provide a medium for follow-up co-culture studies using specific methanogens and Geobacter spp. dominated biofilm anodes. All three methanogens showed growth and activity in the BFS01 medium. This was demonstrated by estimating the specific growth rates (μ) and doubling times (td) of each methanogen. The μ and td based on methane accumulation in the headspace showed values consistent with literature values for M. barkeri and M. soehngenii. However, μ and td based on methane accumulation in the headspace differed from literature data for M. formicicum but still allowed sufficient growth. The lowered salt concentration and the omission of chemically complex organic components in the medium may have led to the observed deviation from μ and td for M. formicicum as well as the changed morphology. 16S rRNA gene-based amplicon sequencing and whole genome nanopore sequencing further confirmed purity and species identity.

The combination of anaerobic digestion (AD) and microbial electrochemical technologies (METs) offers different opportunities to increase the efficiency and sustainability of AD processes. However, methanogenic archaea and/or particles may partially hinder combining MET and AD processes. Furthermore, it is unclear if the applied anode potential affects the activity and efficiency of electroactive microorganisms in AD-MET combinations as it is described for more controlled experimental conditions. In this study, we confirm that 6-week-old Geobacter spp. dominated biofilms are by far more active and stable in AD-effluents than 3-week-old Geobacter spp. dominated biofilms. Furthermore, we show that the biofilms are twice as active at −0.2 V compared to 0.4 V, even under challenging conditions occurring in AD-MET systems. Paired-end amplicon sequencing at the DNA level using 16S-rRNA and mcrA gene shows that hydrogenotrophic methanogens incorporate into biofilms immersed in AD-effluent without any negative effect on biofilm stability and electrochemical activity.