Syntrophic Acetate-oxidizing Bacteria Research Articles

Carbon monoxide conversion by anaerobic microbiome in a thermophilic trickle bed reactor

Biomethanation offers a promising route for the valorization of synthesis gas, yet significant challenges arise from the limited conversion of carbon monoxide (CO). This study investigated the adaptation of an anaerobic microbiome in a continuous trickle bed reactor (TBR) with CO as the sole carbon and energy source. We evaluated reactor performance and microbial community changes under different CO loading rates and gas retention times (GRT). Optimal performance was achieved at a CO loading rate of 5.16 Nm³ m⁻³ d⁻¹ and a GRT of 60.6 min, resulting in average production rates of 0.99 Nm³ m⁻³ d⁻¹ for CH₄ and 2.55 Nm⁻³ m⁻³ d⁻¹ for CO₂, with an 88 % CO conversion rate. Microbial analysis indicated that the community was dominated by the genus Methanothermobacter, known for its ability to utilize CO as a sole substrate, followed by a co-dominance of syntrophic acetate-oxidizing bacteria Syntrophaceticus. This syntrophic relationship between Methanothermobacter and Syntrophaceticus is expected to be crucial for the efficient CO conversion process. Additionally, the study proposes a two-reactor system for converting synthesis gas to grid-quality methane.

Improving dairy manure hydrolysis and acidification through microbial community restructuring by adaptation to hyperthermophilic conditions

Dairy manure (DM) contributes significantly to greenhouse gas emissions and ecosystem degradation, yet its resistance to biodegradation hinders widespread bioprocessing applications. Lignocellulosic materials in DM pose a particular challenge because of their recalcitrance. Bioprocessing under hyperthermophilic (≥70 °C) conditions potentially offers an advantage over traditional fermentation temperatures due to enhanced activity of enzymes and the kinetics of enzymatic reactions. This can lead to a higher conversion rate and a greater extent of biomass hydrolysis and acidification. To test the validity of this hypothesis, the current study evaluated the efficacy of anaerobic hydrolysis and acidogenic fermentation of DM under mesophilic, thermophilic, and hyperthermophilic conditions. All inocula were adapted to corresponding temperatures but were derived from the same mesophilic source. Hyperthermophilic conditions resulted in superior DM hydrolysis efficiency (53%) compared to mesophilic (34%) and thermophilic (42%) conditions. The hyperthermophilic environment was particularly favorable to the decomposition of crude proteins and hemicellulose, which were reduced by 64% and 54%, respectively. Furthermore, hyperthermophilic fermentation also yielded the highest volatile fatty acid (VFA) production rate of 460 mg/L/day during the first four days, representing improvements of 50% and 90% over mesophilic and thermophilic conditions. In part, this was attributed to the enhanced production of branched-chain VFAs, including an increase of 6–10% in isobutyric acid and 12–13% in isovaleric acid. At hyperthermophilic conditions, however, there was no accumulation of VFAs during the days 5–8 of fermentation, which could be due to acetate conversion by the syntrophic acetate-oxidizing bacteria. A considerable gain in hydrolysis efficiency and VFA production rate were accompanied by a reduction in microbial diversity, which suggests that hyperthermophilic temperature is a favorable environment for the selection of organisms with enhanced DM hydrolysis and fermentation capabilities. A significantly increased relative abundance of xylanolytic Caldicoprobacter (23% of population) and proteolytic Thermovirga (9% of population) could be the major contributors to improved decomposition of hemicellulose and protein. As revealed by the techno-economic analysis, acidogenic fermentation of DM at 70 °C and a retention period of 4 days provides the greatest positive net present value, highest internal rate of return of 9.2%, and shortest investment payback period of 9 years. This study demonstrates that hyperthermophilic conditions enable superior deconstruction and bioconversion of lignocellulose-containing biomass into VFAs under reduced retention times, offering a promising approach for improving DM management and generating bioproducts.

Metagenomic insights into the enhanced methane production by hydrochar at varied propionate concentrations

Propionate is a crucial intermediate product in anaerobic digestion (AD), but its degradation is thermodynamically unfavorable. Its accumulation in the AD system can significantly reduce methane production efficiency and may lead to severe acidification. The present study investigated the methane production at various propionate concentrations (15, 30 and 60 mM) with and without hydrochar and also explored the microbial shifts by metagenomic analysis. The findings revealed that higher propionate concentrations reduced methane production and propionate degradation efficiency, leading to acetate accumulation. The addition of hydrochar mitigated the inhibitory effects of increased propionate concentrations, notably reduced the lag phase by 20.13 % and enhanced the methane production rate by 138.43 % under 30 mM propionate. This also resulted in faster degradation of propionate and acetate. Hydrochar impacted the microbial communities’ functional potentials, enhancing methane metabolic potential in all conditions. Genomic-centric analysis revealed that hydrochar selectively enriched syntrophic propionate-oxidizing bacteria (SPOB) candidate MAG79 Syntrophopropionicum sp., which could participate in DIET, across different concentrations of propionate, with the most pronounced effect at 30 mM propionate. This was distinct from the dominant species in the control groups, MAG63 Syntrophosphaera sp. and MAG69 Syntrophobacteraceae sp.. Hydrochar further supported the growth of another DIET candidate MAG4 Methanosarcina mazei and increased the abundance of proposed syntrophic acetate-oxidizing bacterium (SAOBs), across all propionate concentrations. These effects collectively led to more efficient propionate degradation and methane production.

Metabolism of novel potential syntrophic acetate-oxidizing bacteria in thermophilic methanogenic chemostats.

Acetate is a major intermediate in the anaerobic digestion of organic waste to produce CH4. In methanogenic systems, acetate degradation is carried out by either acetoclastic methanogenesis or syntrophic degradation by acetate oxidizers and hydrogenotrophic methanogens. Due to challenges in the isolation of syntrophic acetate-oxidizing bacteria (SAOB), the diversity and metabolism of SAOB and the mechanisms of their interactions with methanogenic partners are not fully characterized. In this study, the in situ activity and metabolic characteristics of potential SAOB and their interactions with methanogens were elucidated through metagenomics and metatranscriptomics. In addition to the reported SAOB classified in the genera Tepidanaerobacter, Desulfotomaculum, and Thermodesulfovibrio, we identified a number of potential SAOB that are affiliated with Clostridia, Thermoanaerobacteraceae, Anaerolineae, and Gemmatimonadetes. The potential SAOB possessing the glycine-mediated acetate oxidation pathway dominates SAOB communities. Moreover, formate appeared to be the main product of the acetate degradation by the most active potential SAOB. We identified the methanogen partner of these potential SAOB in the acetate-fed chemostat as Methanosarcina thermophila. The dominated potential SAOB in each chemostat had similar metabolic characteristics, even though they were in different fatty-acid-fed chemostats. These novel syntrophic lineages are prevalent and may play critical roles in thermophilic methanogenic reactors. This study expands our understanding of the phylogenetic diversity and in situ biological functions of uncultured syntrophic acetate degraders and presents novel insights into how they interact with methanogens.IMPORTANCECombining reactor operation with omics provides insights into novel uncultured syntrophic acetate degraders and how they perform in thermophilic anaerobic digesters. This improves our understanding of syntrophic acetate degradation and contributes to the background knowledge necessary to better control and optimize anaerobic digestion processes.

Adaptation of Anaerobic Digestion Microbial Communities to High Ammonium Levels: Insights from Strain-Resolved Metagenomics.

Ammonia release from proteinaceous feedstocks represents the main inhibitor of the anaerobic digestion (AD) process, which can result in a decreased biomethane yield or even complete failure of the process. The present study focused on the adaptation of mesophilic AD communities to a stepwise increase in the concentration of ammonium chloride in synthetic medium with casein used as the carbon source. An adaptation process occurring over more than 20 months allowed batch reactors to reach up to 20 g of NH4+ N/L without collapsing in acidification nor ceasing methane production. To decipher the microbial dynamics occurring during the adaptation and determine the genes mostly exposed to selective pressure, a combination of biochemical and metagenomics analyses was performed, reconstructing the strains of key species and tracking them over time. Subsequently, the adaptive metabolic mechanisms were delineated by following the single nucleotide variants (SNVs) characterizing the strains and prioritizing the associated genes according to their function. An in-depth exploration of the archaeon Methanoculleus bourgensis vb3066 and the putative syntrophic acetate-oxidizing bacteria Acetomicrobium sp. ma133 identified positively selected SNVs on genes involved in stress adaptation. The intraspecies diversity with multiple coexisting strains in a temporal succession pattern allows us to detect the presence of an additional level of diversity within the microbial community beyond the species level.

Iron-carbon micro-electrolysis material enhanced high-solid anaerobic digestion: Performance and microbial mechanism

Iron-carbon micro-electrolysis (ICME) materials are widely used as composite conductive materials for wastewater treatment to promote the degradation of volatile solids (VS), but there is virtually no research on their effect on high-solid anaerobic digestion (HSAD). This study aimed to investigate the effect of 10–50 g/L ICME dosages on HSAD and to reveal the microbial enhancement mechanism. The batch experiments (working volume of 400 mL) were conducted in a constant temperature water bath shaker. The results showed that the ICME materials promoted methane production by HSAD. The highest methane production rate was 297.79 mL/ g VS at a dosage of 50 g/L, which was 28% higher than that of the control. Scanning electron microscopy results showed that the ICME materials increased the attachment area of microorganisms. Further analyses of microbial community indicated that ICME materials facilitate symbiotic metabolism of syntrophic acetate-oxidizing bacteria (Synergistaceae) and hydrogenotrophic methanogens (Methanospirillum). This potentially promotes the conversion of the acetoacetic methanogenesis pathway to the syntrophic acetate oxidation and hydrogenotrophic methanogenesis pathways. This study revealed ICME material enhancement in HSAD as well as new insights into the microbial mechanisms.

Diverse electron carriers drive syntrophic interactions in an enriched anaerobic acetate-oxidizing consortium

In many anoxic environments, syntrophic acetate oxidation (SAO) is a key pathway mediating the conversion of acetate into methane through obligate cross-feeding interactions between SAO bacteria (SAOB) and methanogenic archaea. The SAO pathway is particularly important in engineered environments such as anaerobic digestion (AD) systems operating at thermophilic temperatures and/or with high ammonia. Despite the widespread importance of SAOB to the stability of the AD process, little is known about their in situ physiologies due to typically low biomass yields and resistance to isolation. Here, we performed a long-term (300-day) continuous enrichment of a thermophilic (55 °C) SAO community from a municipal AD system using acetate as the sole carbon source. Over 80% of the enriched bioreactor metagenome belonged to a three-member consortium, including an acetate-oxidizing bacterium affiliated with DTU068 encoding for carbon dioxide, hydrogen, and formate production, along with two methanogenic archaea affiliated with Methanothermobacter_A. Stable isotope probing was coupled with metaproteogenomics to quantify carbon flux into each community member during acetate conversion and inform metabolic reconstruction and genome-scale modeling. This effort revealed that the two Methanothermobacter_A species differed in their preferred electron donors, with one possessing the ability to grow on formate and the other only consuming hydrogen. A thermodynamic analysis suggested that the presence of the formate-consuming methanogen broadened the environmental conditions where ATP production from SAO was favorable. Collectively, these results highlight how flexibility in electron partitioning during SAO likely governs community structure and fitness through thermodynamic-driven mutualism, shedding valuable insights into the metabolic underpinnings of this key functional group within methanogenic ecosystems.

New attempts on acidic anaerobic digestion of poly (butylene adipate-co-terephthalate) wastewater in upflow anaerobic sludge blanket reactor

Poly (butylene adipate-co-terephthalate) (PBAT) wastewater is a highly concentrated, acidic, and toxic wastewater generated from biodegradable plastics production. Large amounts of alkali would be consumed when treating PBAT wastewater by anaerobic digestion due to the low pH value. This study employed acidic anaerobic digestion to treat PBAT wastewater and compared to neutral anaerobic digestion. The results indicated that the COD removal rates in the acidic upflow anaerobic sludge blanket (UASB) reactor were 65.5% and 59.9%, respectively at influent pH 6.0 and 5.0 with the COD concentration of around 11,000 mg L−1, and the methane conversion efficiency were 172.5 and 183.8 mLCH4/gCODr (gCODr: COD removed amount), respectively. Correspondingly, the average COD removal and methane conversion efficiency in the neutral UASB reactor were 63.2% and 188.0 mLCH4/gCODr, respectively. The treatment efficiency of acidic and neutral UASB reactors for PBAT wastewater was similar. The hydrogenotrophic methanogenic activity was further enhanced in the acidic UASB reactor compared to the neutral one. The increase of alkalinity in the acidic UASB reactor (2.4 mmol L−1) was higher than the neutral (2.0 mmol L−1). A higher level of syntrophic acetate oxidation bacteria and hydrogenotrophic methanogen was enriched in the acidic UASB reactor, ensuring efficient treatment and saving costs.

Presence and role of viruses in anaerobic digestion of food waste under environmental variability

BackgroundThe interaction among microorganisms in the anaerobic digestion of food waste (ADFW) reactors lead to the degradation of organics and the recycling of energy. Viruses are an important component of the microorganisms involved in ADFW, but are rarely investigated. Furthermore, little is known about how viruses affect methanogenesis.ResultsThousands of viral sequences were recovered from five full-scale ADFW reactors. Gene-sharing networks indicated that the ADFW samples contained substantial numbers of unexplored anaerobic-specific viruses. Moreover, the viral communities in five full-scale reactors exhibited both commonalities and heterogeneities. The lab-scale dynamic analysis of typical ADFW scenarios suggested that the viruses had similar kinetic characteristics to their prokaryotic hosts. By associating with putative hosts, a majority of the bacteria and archaea phyla were found to be infected by viruses. Viruses may influence prokaryotic ecological niches, and thus methanogenesis, by infecting key functional microorganisms, such as sulfate-reducing bacteria (SRB), syntrophic acetate-oxidizing bacteria (SAOB), and methanogens. Metabolic predictions for the viruses suggested that they may collaborate with hosts at key steps of sulfur and long-chain fatty acid (LCFA) metabolism and could be involved in typical methanogenesis pathways to participate in methane production.ConclusionsOur results expanded the diversity of viruses in ADFW systems and suggested two ways that viral manipulated ADFW biochemical processes.3DQb7S-2dM-gGjfqAhcADyVideo

Complexity of temperature dependence in methanogenic microbial environments.

There is virtually no environmental process that is not dependent on temperature. This includes the microbial processes that result in the production of CH4, an important greenhouse gas. Microbial CH4 production is the result of a combination of many different microorganisms and microbial processes, which together achieve the mineralization of organic matter to CO2 and CH4. Temperature dependence applies to each individual step and each individual microbe. This review will discuss the different aspects of temperature dependence including temperature affecting the kinetics and thermodynamics of the various microbial processes, affecting the pathways of organic matter degradation and CH4 production, and affecting the composition of the microbial communities involved. For example, it was found that increasing temperature results in a change of the methanogenic pathway with increasing contribution from mainly acetate to mainly H2/CO2 as immediate CH4 precursor, and with replacement of aceticlastic methanogenic archaea by thermophilic syntrophic acetate-oxidizing bacteria plus thermophilic hydrogenotrophic methanogenic archaea. This shift is consistent with reaction energetics, but it is not obligatory, since high temperature environments exist in which acetate is consumed by thermophilic aceticlastic archaea. Many studies have shown that CH4 production rates increase with temperature displaying a temperature optimum and a characteristic apparent activation energy (Ea). Interestingly, CH4 release from defined microbial cultures, from environmental samples and from wetland field sites all show similar Ea values around 100 kJ mol-1 indicating that CH4 production rates are limited by the methanogenic archaea rather than by hydrolysis of organic matter. Hence, the final rather than the initial step controls the methanogenic degradation of organic matter, which apparently is rarely in steady state.

Performance of anaerobic digestion of phenol using exogenous hydrogen and granular activated carbon and analysis of microbial community.

Anaerobic conversion rate of phenol to methane was low due to its biological toxicity. In this study, the coupling of granular activated carbon (GAC) and exogenous hydrogen (EH) could enhance greatly methane production of phenol anaerobic digestion, and the metagenomic was firstly used to analyze its potential mechanism. The results indicated that a mass of syntrophic acetate-oxidizing bacteria and hydrogen-utilizing methanogens were enriched on the GAC surface, and SAO-HM pathway has become the dominant pathway. The energy transfer analysis implied that the abundance of adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NADH) oxidase increased. Furthermore, direct interspecies electron transfer (DIET) was formed by promoting type IV e-pili between Methanobacterium and Syntrophus, thereby improving the interspecies electron transfer efficiency. The dominant SAO-HM pathway was induced and DIET was formed, which was the internal mechanism of the coupling of GAC and EH to enhance anaerobic biotransformation of phenol.

Syntrophic Butyrate-Oxidizing Consortium Mitigates Acetate Inhibition through a Shift from Acetoclastic to Hydrogenotrophic Methanogenesis and Alleviates VFA Stress in Thermophilic Anaerobic Digestion

In anaerobic digestion (AD), butyrate is degraded by syntrophic consortium, but can accumulate in highly loaded AD systems. The effect of butyrate on the AD process attracts much less attention than propionate or acetate. In this work, an enrichment culture of the thermophilic butyrate-oxidizing syntrophic consortium was obtained by gradually increasing the initial butyrate concentration from 20 to 170 mM. Surprisingly, even the highest butyrate concentration did not significantly inhibit the methanogenic community, and the stage of acetate degradation was the limiting overall rate of the process. At 170 mM butyrate, the bacterial community changed towards the dominance of syntrophic acetate-oxidizing (SAO) bacteria related to Syntrophaceticus (42.9%), Syntrophomonas (26.2%) and Firmicutes (26.2%), while the archaeal community experienced a sharp decrease in the abundance of Methanosarcina thermophila (from 86.0 to 25.0%) and increase in Methanothermobacter thermautotrophicus (from 3.2 to 53.1%) and Methanomassiliicoccus (from 3.2 to 21.9%). Thus, the shift from acetoclastic methanogenesis to SAO coupled to hydrogenotrophic methanogenesis occurred as an adaptive strategy to overcome high acetate (~200 mM) build-up. Bioaugmentation with the obtained enrichment culture was effective in mitigating the butyrate-dominated VFA build-up during the AD of readily biodegradable waste, increasing the methane production rate, methane yield and volatile solids removal by more than 3.5, 6.2 and 2.9 times, respectively. Our study revealed that the thermophilic butyrate-oxidizing consortia as bioaugmented culture could be the potential strategy to alleviate the high organic load and VFA stress of AD.

Binary-component anaerobic co-digestion: Synergies and microbial profiles

One of the key issues to create synergistic effect in anaerobic co-digestion (AcoD), is how to make the appropriate choice about co-substrate and determine the optimal mixing ratio. Adjusting organic component, namely carbohydrate (C), protein (P) and lipid (L), is expected to be a dependable strategy for obtaining synergistic effect in anaerobic co-digestion. In this study, the AcoD performances of organic components at different load levels were investigated in terms of methane yield, metabolic transformation, and microbial response. The synergistic interval at different loading levels showed considerable stability, and the significant synergy was achieved at C3P7 to C8P2 except for C6P4, C4L6 to C6L4, and P3L7 to P5L5 (based on mass ratio), respectively. Under high feeding load (4%VS), groups within synergistic interval could significantly reduce the toxic effects of intermediate metabolic inhibitors and improve methane production. However, AcoD of protein and lipid would induce antagonistic effects, and high proportion of protein was more serious. In the face of adversity, bacterial community diversity was weakened and the relative abundance of dominated phylum Firmicutes was strengthened. Furthermore, acetoclastic methanogenesis pathway was transformed to hydrogenotrophic pathway since hydrogenotrophic methanogens replaced acetoclastic methanogens as the dominant methanogenic archaea with the increase of syntrophic acetate oxidation bacteria. Suitable organic components proportion can effectively reduce inhibitor accumulation and maintain a well methanogens structure to ensure high methane production.

Granular activated carbon and exogenous hydrogen enhanced anaerobic digestion of hypersaline phenolic wastewater via syntrophic acetate oxidation and hydrogenotrophic methanogenesis

High salinity in phenolic wastewater inhibited anaerobes' metabolic activity, thereby affecting the anaerobic biotransformation of phenol. In this study, granular activated carbon (GAC) coupled with exogenous hydrogen (H2) was used to enhance the anaerobic digestion of phenol. The GAC/H2 group's accumulative methane production, coenzyme F420 concentration, and interspecies electron transfer system activity increased by 24 %, 53 %, and 16 %, respectively, compared with the control group. In the floc sludge of the GAC/H2 group, the relative abundance of syntrophic acetate-oxidizing bacteria such as Syntrophus and Syntrophorhabdus were 18.7 % and 1.1 % at genus level, respectively, which were around 93.5 and 7.5 times of that of the controlgroup. Moreover, Acinetobacter (77.6 %), Methanobacterium (44.0 %), and Methanosarcina (34.2 %) were significantly enriched on the GAC surface in GAC/H2 group. Therefore, the coupling of GAC and H2 provided a novel attempt at anaerobic digestion of hypersaline phenolic wastewater via syntrophic acetate oxidation and hydrogenotrophic methanogenesis pathway.

Effect of nano-Fe3O4 biochar on anaerobic digestion of chicken manure under high ammonia nitrogen concentration

In order to improve the stability and CH4 production efficiency of the anaerobic digestion (AD) system under high ammonia nitrogen (AN) concentration, this study investigated the effect of biochar loaded with nano-Fe3O4 particles to generate nano-Fe3O4 biochar on AD of chicken manure (CM). The results showed that biochar can be used as an excellent carrier for nano-Fe3O4 particles, solving the problem of easy agglomeration and loss. The inhibition of the AD system caused by high concentrations of AN was greatly mitigated after loading nano-Fe3O4 particles on biochar, and the CH4 yield was substantially increased. The highest CH4 yield was achieved when the amount of nano-Fe3O4 biochar was 15%, which was 62.61% higher compared to the control group. Microbial community analysis showed that the porous structure of biochar was capable of immobilizing and enriching microorganisms, and the relative abundance of Firmicutes and Methanobacterium increased by 28.67–54.44% and 52.62–87.70%, respectively. Firmicutes acted as an important acetate bacterium, contributing to syntrophic acetate oxidation (SAO) with hydrotropic methanogenesis. SAO was an effective alternative pathway for acetate degradation under ammonia stress involving the syntrophic activities of SAO bacteria and hydrogenotrophic archaea. The load of nano-Fe3O4 particles increased the relative abundance of Chloroflexi from 0.52% to 3.51% and the Methanosarcina from 27.50% to 49.30%. Chloroflexi and Methanosarcina could act as potential direct electron transfer (DIET) partners for facilitating methane production by promoting the SAO pathway.

Syntrophic Acetate-Oxidizing Microbial Consortia Enriched from Full-Scale Mesophilic Food Waste Anaerobic Digesters Showing High Biodiversity and Functional Redundancy.

ABSTRACTSyntrophic acetate oxidation (SAO) coupled with hydrogenotrophic methanogenesis (HM) plays a vital role in the anaerobic digestion of protein-rich feedstocks such as food wastes. However, current knowledge of the biodiversity and genetic potential of the involved microbial participants, especially syntrophic acetate-oxidizing bacteria (SAOB), is limited due to the low abundance of these microorganisms and challenges in their isolation. The intent of this study was to enrich and identify potential SAOB. Therefore, we conducted continuous acetate feeding under high ammonia concentrations using two separate inoculum consortia of microorganisms that originated from full-scale mesophilic food waste digesters, which lasted for more than 200 days. Using 16S rRNA gene amplicon and metagenomic analyses, we observed a convergence of the experimental microbial communities during the enrichment regarding taxonomic composition and metabolic functional composition. Stable carbon isotope analyses of biogas indicated that SAO-HM was the dominant methanogenic pathway during the enrichment process. The hydrogenotrophic methanogen Methanoculleus dominated the archaeal community. The enriched SAO community featured high biodiversity and metabolic functional redundancy. By analyzing the metagenome-assembled genomes, the known SAOB Syntrophaceticus schinkii and six uncultured populations were identified to have the genetic potential to perform SAO through the conventional reversed Wood-Ljungdahl pathway, while another six bacteria were found to encode the reversed Wood-Ljungdahl pathway combined with a glycine cleavage system as novel SAOB candidates. These results showed that the food waste anaerobic digesters harbor diverse SAOB and highlighted the importance of the glycine cleavage system for acetate oxidation.IMPORTANCE Syntrophic acetate oxidation to CO2 and H2, together with hydrogenotrophic methanogenesis, contributes to much of the carbon flux in the anaerobic digestion of organic wastes, especially at high ammonia concentrations. A deep understanding of the biodiversity, metabolic genetic potential, and ecology of the SAO community can help to improve biomethane production from wastes for clean energy production. Here, we enriched the SAO-HM functional guild obtained from full-scale food waste anaerobic digesters and recorded dynamic changes in community taxonomic composition and functional profiles. By reconstructing the metabolic pathways, diverse known and novel bacterial members were found to have SAO potential via the reversed Wood-Ljungdahl (WL) pathway alone, or via the reversed WL pathway with a glycine cleavage system (WLP-GCS), and those catalyzing WLP-GCS showed higher microbial abundance. This study revealed the biodiversity and metabolic functional redundancy of SAOB in full-scale anaerobic digester systems and provided inspiration for further genome-centric studies.

Exploring key factors in anaerobic syntrophic interactions: Biomass activity, microbial community, and morphology

The present work evaluated the impacts of microbial communities, biomass activity and sludge morphology on anaerobic syntrophic reactions. Experiments were conducted using mature floc sludge and granular sludge under different food/microbes ratios, and with different sludge types (floc sludge, concentrated floc sludge and granular sludge) and sludge morphology (granules, vortexed granules, and granules with different particle sizes). The results show that the intact granules achieved the most effective syntrophic reaction among all sludge types. The granule structure facilitated the enrichment of syntrophic acetate oxidation bacteria (g_Syner-01 and g_Mesotoga) and methanogens, which corresponds to their superior specific methanogenic activity and high production of communication compounds. Despite the high diffusion and substrate uptake capacities, the disintegrated granules had low H2 consumption rates, which led to poor syntrophic activities. The results underline the importance of sludge spatial structures in promoting excellent syntrophic activities and the development of diverse microbial communities.

Serial anaerobic digestion improves protein degradation and biogas production from mixed food waste

Optimization of the biogas generation process is important to achieve efficient degradation and high methane yield, and to reduce methane emissions from the digestate. In this study, serial digester systems with two or three biogas reactors were compared with a single reactor, with the aim of improving degree of degradation and methane yield from food waste and assessing adaptation of microbial communities to different reactor steps. All systems had the same total organic load (2.4 g VS/(L d)) and hydraulic retention time (55 days). Serial systems increased methane yield by >5% compared with the single reactor, with the majority of the methane being obtained from the first-step reactors. Improved protein degradation was also obtained in serial systems, with >20% lower outgoing protein concentration compared with the single reactor and increasing NH4+-N concentration with every reactor step. This resulted in separation of high ammonia (>384 mg NH3-N/L) levels from the main methane production, reducing the risk of methanogen inhibition. Methanosarcina dominated the methanogenic community in all reactors, but increases in the hydrogenotrophic genera Methanoculleus and Methanobacterium were observed at higher ammonia levels. Potential syntrophic acetate-oxidizing bacteria, such as MBA03 and Dethiobacteraceae, followed the same trend as the hydrogenotrophic methanogens. Phylum Bacteroidota family Paludibacteraceae was highly abundant in the first steps and then decreased abruptly, potentially linked to an observed decrease in degradation in the last-step reactors. Nevertheless, the results indicated a trend of increasing relative abundance of the potentially proteolytic genera Proteiniphilum and Fastidiosipila with successive reactor steps.

Diverse acetate-oxidizing syntrophs contributing to biogas production from food waste in full-scale anaerobic digesters in China

Anaerobic digestion is a sustainable biotechnology for treating food waste (FW) with biogas production driven by microorganisms. Generation of FW increased greatly in China, which are mainly treated with anaerobic digestion technology. A survey into nine full-scale mesophilic anaerobic FW digesters was conducted, together with four sewage sludge digesters as a comparison, to investigate the specific physicochemical features and the resulted microbiota. Results showed that FW digesters were characterized with high salt, volatile fatty acid and ammonia concentrations. A unique “core” microbial community indigenous inside FW digesters was identified, including 70 bacterial and 4 archaeal genus-level taxa, which was quite different from that in sludge digesters. Stable carbon isotope signature of biogas revealed that, hydrogenotrophic methanogenesis (HM) played a significant role in methane generation. The hydrogenotrophic Methanobacterium demonstrated strong positive correlations with the syntrophic acetate oxidizing bacteria (SAOB) Syntrophaceticus and other SAOB candidates (Aminobacterium, Gelria, DTU014, Alkaliphilus), forming a unique SAO-HM functional guild which particularly prevailed in digesters receiving household FW. Feedstock composition was the essential factor influencing microbial structure, followed by salinity, ammonia, and VFAs. The high ammonia concentration and prominent contribution of diverse SAO-HM players indicate that, tailored management strategy needs to be developed for treating household FW.

The synergistic effect of rumen cellulolytic bacteria and activated carbon on thermophilic digestion of cornstalk

To explore the bioaugmentation of rumen cellulolytic bacteria (RCB) and activated carbon (AC) on thermophilic digestion of cornstalk, biochemical methane potential tests were carried out. Adding RCB or AC can improve methane production, while simultaneous existence of AC (10 g/L) and RCB (5%) obtained the best performance. The maximum cellulose degradation rate, methane production rate and methane yield were 66.92%, 32.2 L/(kgVS·d), and 144.9 L/kgVS, which increased by 30.23%, 51.17%, and 20.35% compared with control group. The cellulolytic and fermentative bacteria (Hydrogenispora), syntrophic acetate-oxidizing bacteria (norank_o_MBA03), and hydrogenotrophic Methanothermobacter were crucial for thermophilic digestion of cornstalk. The enhancement of AC was due to the enrichment of Hydrogenispora and Methanothermobacter, while RCB can increase the abundance of cellulolytic bacteria (Halocella and norank_o_M55-D21) and mixotrophic Methanosarcina. The synergetic effect of AC and RCB owing to the enriched cellulolytic bacteria, the enhanced syntrophic acetate oxidation and the concentrated carbon metabolic flow to methane.