Tailoring a highly conductive and super-hydrophilic electrode for biocatalytic performance of microbial electrolysis cells.

Bioelectrochemical hydrogen production via microbial electrolysis cells (MECs) has attracted attention as the next generation of technology for the hydrogen economy. MECs work by electrochemically active bacteria reducing organic compounds at the anode. However, the hydrophobic nature of carbon-based anodes suppresses the release of the produced gas and water penetration, which significantly reduces the possibility of microbial attachment. Consequently, a limited surface area of the anode is used, which decreases hydrogen production efficiency. In this study, the bifunctional material poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) was applied to the surface of a three-dimensional carbon felt anode to enhance the hydrogen production efficiency of an MEC owing to the high conductivity of PEDOT and super-hydrophilicity of PSS. In experiments, the PEDOT:PSS-modified anode almost doubled the hydrogen production efficiency of the MEC compared with the control anode owing to the increased capacitance current (239.3 %) and biofilm formation (220.7 %). The modified anode reduced the time required for the MEC to reach a steady state of hydrogen production by 14 days compared to the control anode. Microbial community profiles demonstrated that the modified anode had a greater abundance of electrochemically active bacteria than the control anode. This simple method could be widely applied to various bioelectrochemical systems (e.g., microbial fuel cells and solar cells) and to scaling up MECs.

Key players in syntrophic propionate oxidation revealed by metagenome-assembled genomes from anaerobic digesters bioaugmented with propionic acid enriched microbial consortia.

Propionic acid (HPr) is frequently accumulated in anaerobic digesters due to its thermodynamically unfavorable degradation reaction. Here, we identify key players in HPr oxidation and organic overloading recovery from metagenome-assembled genomes (MAGs) recovered from anaerobic digesters inoculated with HPr-enriched microbial consortia before initiating organic overloading. Two independent HPr-enrichment cultures commonly selected two uncultured microorganisms represented with high relative abundance: Methanoculleus sp002497965 and JABUEY01 sp013314815 (a member of the Syntrophobacteraceae family). The relative abundance of JABUEY01 sp013314815 was 60 times higher in bioaugmented bioreactors compared to their unaugmented counterparts after recovery from organic overloading. Genomic analysis of JABUEY01 sp013314815 revealed its metabolic potential for syntrophic propionate degradation when partnered with hydrogenotrophic methanogens (e.g., Methanoculleus sp002497965) via the methylmalonyl-CoA pathway. Our results identified at least two key species that are responsible for efficient propionate removal and demonstrate their potential applications as microbial cocktails for stable AD operation.

Complex network analysis of slaughterhouse waste anaerobic digestion: From failure to success of long-term operation.

The study explored slaughterhouse waste (SHW) as prime feedstock associated with and without supplement of an external slowly degradable lignocellulosic carbon source to overcome the synergistic co-inhibitions of ammonia and fatty acids. Long-term solid-state digestion (SSD) and liquid-state digestion (LSD) were investigated using a mixture of pork liver and fat. At 2.0 g volatile solids (VS) L-1 d-1 of organic loading rate (OLR), the two reactors of SSD experienced operational instability due to ammonia inhibition and volatile fatty acid (VFA) accumulation while LSD successfully produced 0.725 CH4 L CH4 g-1VS during 197 d of working days under unfavorable condition with high total ammonia nitrogen (>4.7 g/L) and VFAs concentration (>1.9 g/L). The network analysis between complex microflora and operational parameters provided an insight for sustainable biogas production using SHW. Among all, hydrogenotrophic methanogens have shown better resistance than acetoclastic methanogens.

Effects of Different pH Control Strategies on Microalgae Cultivation and Nutrient Removal from Anaerobic Digestion Effluent.

This study investigated nutrient removal from anaerobic digestion effluent by cultivating mixed-culture microalgae enriched from anaerobic sludge under different pH conditions: RUC (uncontrolled), R7-8 (maintained at 7-8), and R<8 (maintained below 8). Significant amounts of NH4+-N were lost by volatilization in RUC cultures due to increased pH values (≤8.6) during the early period of cultivation. The pH control strategies significantly affected the biological NH4+-N removal (highest in R7-8), microalgal growth (highest in R7-8), biomass settleability (highest in R<8), and microalgal growth relative to bacteria (highest in R<8) in the cultures. Parachlorella completely dominated the microalgal communities in the inoculum and all of the cultures, and grew well at highly acidic pH (<3) induced by culture acidification with microalgal growth. Microalgae-associated bacterial community structure developed very differently among the cultures. The findings call for more attention to the influence and control of pH changes during cultivation in microalgal treatment of anaerobic digestion effluent.

Transcriptomic Analysis of Degradative Pathways for Azo Dye Acid Blue 113 in Sphingomonas melonis B-2 from the Dye Wastewater Treatment Process.

BACKGROUND: Acid Blue 113 (AB113) is a typical azo dye, and the resulting wastewater is toxic and difficult to remove. METHODS: The experimental culture was set up for the biodegradation of the azo dye AB113, and the cell growth and dye decolorization were monitored. Transcriptome sequencing was performed in the presence and absence of AB113 treatment. The key pathways and enzymes involved in AB113 degradation were found through pathway analysis and enrichment software (GO, EggNog and KEGG). RESULTS: S. melonis B-2 achieved more than 80% decolorization within 24 h (50 and 100 mg/L dye). There was a positive relationship between cell growth and the azo dye degradation rate. The expression level of enzymes involved in benzoate and naphthalene degradation pathways (NADH quinone oxidoreductase, N-acetyltransferase and aromatic ring-hydroxylating dioxygenase) increased significantly after the treatment of AB113. CONCLUSIONS: Benzoate and naphthalene degradation pathways were the key pathways for AB113 degradation. NADH quinone oxidoreductase, N-acetyltransferase, aromatic ring-hydroxylating dioxygenase and CYP450 were the key enzymes for AB113 degradation. This study provides evidence for the process of AB113 biodegradation at the molecular and biochemical level that will be useful in monitoring the dye wastewater treatment process at the full-scale treatment.

Long-term effectiveness of bioaugmentation with rumen culture in continuous anaerobic digestion of food and vegetable wastes under feed composition fluctuations.

Biogas plants treating food waste (FW) often experience feed load and composition fluctuations. In Korea, vegetable waste from the preparation of kimchi comprises over 20% of the total FW production during the Kimjang season. The large production of Kimjang waste (KW) can cause mechanical and operational problems in FW digesters. This study investigated the long-term effectiveness of bioaugmentation with rumen culture (38 months) in an anaerobic reactor co-digesting FW with varying amounts of KW. The bioaugmented reactor maintained better and stabler performance under recurrent fluctuations in feed characteristics than a non-bioaugmented control reactor, particularly under high ammonia conditions. Bioaugmentation increased microbial diversity, thereby improving the resilience of the microbial community. Some augmented microorganisms, especially Methanosarcina, likely played an important role in it. The results suggest that the proposed bioaugmentation strategy may provide a means to effectively treat and valorize KW-and potentially other seasonal lignocellulosic wastes-by co-digestion with FW.

Tracking microbial community shifts during recovery process in overloaded anaerobic digesters under biological and non-biological supplementation strategies.

Anaerobic digestion encounters operational instability due to fluctuations in organic loading. Propionic acid (HPr) is frequently accumulated due to its unfavorable reaction thermodynamics. Here, 'specific' bioaugmentation using HPr enrichment cultures (three different injection regimes of quantity and frequency) was compared with 'non-specific' bioaugmentation using anaerobic sludge, and with non-biological supplementation of magnetite or coenzyme M. The specific bioaugmentation treatments showed superior recovery responses during continuous feeding after a peak overload. A 'one-shot' bioaugmentation with enrichment showed the best remediation, with ~25% recovery time and >10% CH4 conversion efficiency compared to the control. Consecutive bioaugmentation showed evidence of increased stability of the introduced community. Families Synergistaceae, Syntrophobacteraceae, and Kosmotogaceae were likely responsible for HPr-oxidation, in potential syntrophy with Methanoculleus and Methanobacterium. The different supplementation strategies can be considered to reduce the effect of start-up or overload in anaerobic digesters based on the availability of supplementation resources.

Density profile modeling for real-time estimation of liquid level in anaerobic digester using multiple pressure meters.

The liquid level of a bioreactor is an important operating parameter governing the hydraulic retention time. In this study, a novel method is proposed to estimate the liquid level of anaerobic digesters. The proposed method has an advantage over typical differential pressure measurement as it considers the heterogeneity of the digestate along the level using multiple pressure meters. The real-time measurement generates a model to fit the densities at different liquid columns, predicts the density of the surface layer and determines the overall liquid level. A pilot-scale (0.33 m3 working volume; 1.2 m liquid level) digester, equipped with seven pressure meters, was operated to test the methodology. The performance of the digester was confirmed stable during a long-term (175 d) operation. A set of density-pressure models was developed and were validated using the long-term experimental data. The new method employing cubic model showed significantly better estimation of the reactor level (mean error rate of 1.31%) with improved CDF, as compared with the traditional differential pressure method (mean error rate of 5.71%). The methodology proposed in this study is simple, robust, and cost-effective and can be used to provide additional insights into the operation of an anaerobic digester such as assessing the mixing efficiency.

Determining the composition of bacterial community and relative abundance of specific antibiotics resistance genes via thermophilic anaerobic digestion of sewage sludge.

In this study, the effects of different temperature transitions on the dynamics of antibiotic resistance genes (ARGs) and bacterial community were investigated during start-up of thermophilic anaerobic digestion (AD) of sewage sludge. Although two thermophilic reactors showed dissimilar removal efficiencies of ARGs in batch mode, both the removal efficiency and reduction patterns of ARGs were similar in continuous stirred tank reactor (CSTR) mode, resulting in significant reduction of the total sum of the relative abundance of ARGs. Using network analysis to explore the correlation between bacterial community and some specific ARGs revealed that composition of the bacterial community played a vital role in the fluctuations in the relative abundance of the antibiotic resistome, demonstrating that shaping the development of ARGs was facilitated by vertical gene transfer. To facilitate eliminating ARGs, minimizing their hosts which persist even under long-term operations is vital in thermophilic AD.

Non-sterile fermentation of food waste with indigenous consortium and yeast - Effects on microbial community and product spectrum.

This work presents examples of non-sterile mixed culture fermentation of food waste with a cultivated indigenous consortium (IC) gained from food waste, which produces lactic and acetic acids, combined with Saccharomyces cerevisiae, which produces ethanol. All results are flanked by microbial analysis to monitor changes in microbial community. At pH 6 and inoculated with yeast or IC, or both mixed sugars conversion was equal to 71%, 51%, or 67%, respectively. Under pH unregulated conditions metabolic yields were 71%, 67%, or up to 81%. While final titer of acetic acid was not affected by pH (100-200 mM), ethanol and lactic acid titers were. Using mixed culture and pH 6, sugars were almost equally used for formation of ethanol and lactic acid (400-500 mM). However, under pH unregulated conditions 80% of the substrate was converted into ethanol (900-1000 mM).

Metagenomic analysis of relationships between the denitrification process and carbon metabolism in a bioaugmented full-scale tannery wastewater treatment plant.

The goal of this study was to investigate the relationship between the denitrification process and carbon metabolism in a full-scale tannery wastewater treatment plant bioaugmented with the microbial consortium BM-S-1. The metagenomic analysis of the microbial community showed that Brachymonas denitrificans, a known denitrifier, was present at a high level in the treatment stages of buffering (B), primary aeration (PA), and sludge digestion (SD). The occurrences of the amino acid-degrading enzymes alpha ketoglutarate dehydrogenase (α-KGDH) and tryptophan synthase were highly correlated with the presence of denitrification genes, such as napA, narG, nosZ and norB. The occurrence of glutamate dehydrogenase (GDH) was also highly paralleled with the occurrence of denitrification genes such as napA, narG, and norZ. The denitrification genes (nosZ, narG, napA, norB and nrfA) and amino acid degradation enzymes (tryptophan synthase, α-KGDH and pyridoxal phosphate dependent enzymes) were observed at high levels in B. This indicates that degradation of amino acids and denitrification of nitrate may potentially occur in B. The high concentrations of the fatty acid degradation enzyme groups (enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase and ß-ketothiolase) were observed together with the denitrification genes, such as napA, narG and nosZ. Phospholipase/carboxylesterase, enoyl-CoA hydratase/isomerase, acyl-CoA dehydrogenase, phenylacetate degradation enzyme and 3-hydroxyacyl-CoA dehydrogenase 2 were also dominant in B. All these results clearly indicate that the denitrification pathways are potentially linked to the degradation pathways of amino acids and fatty acids whose degradation products go through the TCA cycle, generating the NADH that is used as electron donors for denitrification.

Methanogenesis stimulation and inhibition for the production of different target electrobiofuels in microbial electrolysis cells through an on-demand control strategy using the coenzyme M and 2-bromoethanesulfonate.

Electron allocation through the suppression or the stimulation of methanogenesis is critical for microbial electrolysis cells (MECs) to produce the desired target product (e.g., CH4 or H2). In this study, selective methanogenesis control using the coenzyme M (CoM) and 2-bromoethanesulfonate (2-BES) was investigated in a two-chambered MEC to evaluate the effect of CoM and 2-BES on the production of different electrobiofuels, net energy conversion efficiency and microbial community structure. Because the CoM is a crucial methyl-group carrier in the final process of methanogenesis, it was postulated that CoM would stimulate methanogenic activity at the anode, while a structural analog of the CoM (i.e., 2-BES) was expected to improve cathodic H2 yield using electrons conserved because of methanogen inhibition (electron equivalence: 8 mol e- = 1 mol CH4 = 4 mol H2). CoM injection in MECs significantly enhanced their CH4 production rate, purity, and yield by 4.5-fold, 14.5%, and 76.1%, respectively, compared to the control. Moreover, microbial community analysis indicated that Methanosaeta, the major acetoclastic methanogen, continued to dominate the microbial community but steadily decreased in relative abundance after the CoM injection. On the other hand, drastic increases in hydrogenotrophic methanogens, such as Methanoculleus and Methanolinea, were observed along with potential syntrophic acetate-oxidizing bacteria. In contrast, CH4 production in the 2-BES injected trials was significantly inhibited by 79.5%, resulting in a corresponding increase of H2 production by 145.5% compared to the control. Unlike the CoM, the microbial community did not noticeably change when 2-BES was injected, although the population size gradually decreased over time. Also, a single injection of CoM and 2-BES, even at low concentrations (500 µM), enabled the desired allocation of electrons as characterized by a high sensitivity, fast response, and negligible interference. In terms of energy conversion efficiency, methanogenesis stimulation approach resulted in higher net energy production than inhibition approach, whereas the remained electrons were not fully converted to hydrogen in case of the inhibition trial, thus producing less energy.