Hydrogenotrophic Methanogenic Activity Research Articles

Impact of additives on syntrophic propionate and acetate enrichments under high-ammonia conditions

High ammonia concentrations in anaerobic degradation systems cause volatile fatty acid accumulation and reduced methane yield, which often derive from restricted activity of syntrophic acid-oxidising bacteria and hydrogenotrophic methanogens. Inclusion of additives that facilitate the electron transfer or increase cell proximity of syntrophic species by flocculation can be a suitable strategy to counteract these problems, but its actual impact on syntrophic interactions has yet to be determined. In this study, microbial cultivation and molecular and microscopic analysis were performed to evaluate the impact of conductive (graphene, iron oxide) and non-conductive (zeolite) additives on the degradation rate of acetate and propionate to methane by highly enriched ammonia-tolerant syntrophic cultures derived from a biogas process. All additives had a low impact on the lag phase but resulted in a higher rate of acetate (except graphene) and propionate degradation. The syntrophic bacteria ‘Candidatus Syntrophopropionicum ammoniitolerans’, Syntrophaceticus schinkii and a novel hydrogenotrophic methanogen were found in higher relative abundance and higher gene copy numbers in flocculating communities than in planktonic communities in the cultures, indicating benefits to syntrophs of living in close proximity to their cooperating partner. Microscopy and element analysis showed precipitation of phosphates and biofilm formation in all batches except on the graphene batches, possibly enhancing the rate of acetate and propionate degradation. Overall, the concordance of responses observed in both acetate- and propionate-fed cultures highlight the suitability of the addition of iron oxide or zeolites to enhance acid conversion to methane in high-ammonia biogas processes.Key points• All additives promoted acetate (except graphene) and propionate degradation.• A preference for floc formation by ammonia-tolerant syntrophs was revealed.• Microbes colonised the surfaces of iron oxide and zeolite, but not graphene.Graphical

The Effect of Bismuth and Tin on Methane and Acetate Production in a Microbial Electrosynthesis Cell Fed with Carbon Dioxide.

This study investigates the impacts of bismuth and tin on the production of CH4 and volatile fatty acids in a microbial electrosynthesis cell with a continuous CO2 supply. First, the impact of several transition metal ions (Ni2+, Fe2+, Cu2+, Sn2+, Mn2+, MoO42-, and Bi3+) on hydrogenotrophic and acetoclastic methanogenic microbial activity was evaluated in a series of batch bottle tests incubated with anaerobic sludge and a pre-defined concentration of dissolved transition metals. While Cu is considered a promising catalyst for the electrocatalytic conversion of CO2 to short chain fatty acids such as acetate, its presence as a Cu2+ ion was demonstrated to significantly inhibit the microbial production of CH4 and acetate. At the same time, CH4 production increased in the presence of Bi3+ (0.1 g L-1) and remained unchanged at the same concentration of Sn2+. Since Sn is of interest due to its catalytic properties in the electrochemical CO2 conversion, Bi and Sn were added to the cathode compartment of a laboratory-scale microbial electrosynthesis cell (MESC) to achieve an initial concentration of 0.1 g L-1. While an initial increase in CH4 (and acetate for Sn2+) production was observed after the first injection of the metal ions, after the second injection, CH4 production declined. Acetate accumulation was indicative of the reduced activity of acetoclastic methanogens, likely due to the high partial pressure of H2. The modification of a carbon-felt electrode by the electrodeposition of Sn metal on its surface prior to cathode inoculation with anaerobic sludge showed a doubling of CH4 production in the MESC and a lower concentration of acetate, while the electrodeposition of Bi resulted in a decreased CH4 production.

Enhancing energy recovery and microbial stability in blackwater anaerobic digestion with graphite felt

Maximizing energy recovery from blackwater (toilet wastewater) helps the sustainable development of human society. This study introduces a novel approach by integrating graphite felt in anaerobic baffled reactor for enhanced blackwater treatment under room temperature. During the 382-day operation, incorporating graphite felt effectively mitigated hydrolysis and methanogenesis constraints, resulting in a 1.26 to 3.53-fold increase in methane yield compared to the control. The maximum energy recovery was attained at a hydraulic retention time of 1.59 d and an organic loading rate of 2.68 kg COD/m3reactor/d, with a methane production rate of 371 ± 55 NL CH4/m3reactor/d and a methanization rate of 39.44 ± 5.58 % in graphite felt amended reactor. Our findings revealed that graphite felt augmented the efficiency of microbial and enzymatic interactions with the substrate, and enriched hydrolysis fermentation-associated bacteria for efficient solid matter degradation. Further analysis unveiled that graphite felt strengthened the activity of hydrogenotrophic methanogens and fortified the syntrophic relationship between bacteria and archaea. This led to the promotion of a combined syntrophic acetate oxidation and hydrogenotrophic methanogenesis pathway, which guaranteed a stable and efficient methane production process. Overall, this approach offers a cost-effective and efficient strategy to support sustainable development in less-developed, resource-constrained regions.

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.

Diurnal Methane Dynamics in the Cyanobacterial Community of Soda Lake Bitter 1 (Kulunda Steppe, Altai Krai)

Abstract—The diurnal dynamics of methane content in the near-bottom phototrophic biomass of the shallow soda Lake Bitter 1 (Gorchina 1) with a salinity of 30 g/L during the study period was investigated. The community was dominated by filamentous cyanobacteria, although no layered mat was formed. The maximum value of methane content up to 202.4 nmol CH4/cm3 was revealed in the morning hours, which significantly exceeded the night values (28.9–42.8 nmol CH4/cm3). Comparison of data on the content of methane with the relative abundance of cyanobacteria, methanogenic archaea, and methanotrophic bacteria during the twenty-four hours indicated that active processes of the methane cycle in soda lakes occurred not only in the sediments, but also in the near-bottom cyanobacterial communities. Methane content in the biomass of such a community is the result of a balance between the processes of its release by methanogens, consumption by methanotrophic bacteria, and natural degassing. It is assumed that the morning peak of methane content is associated with the release of hydrogen by diazotrophic cyanobacteria, which stimulates the development and activity of hydrogenotrophic methanogens of the genus Methanocalculus.

Feasibility of successive hydrogen and methane production in a single-reactor configuration of batch anaerobic digestion through bioaugmentation and stimulation of hydrogenase activity and direct interspecies electron transfer

Two-stage anaerobic digestion is a promising way to increase the efficiency of decomposition of organic waste. However, spatial separation of stages makes this technology less economically attractive. In this work, we studied a model of two-stage anaerobic digestion with stages separated in time in one reactor. To implement this model, we used a combination of (1) bioaugmentation with a hydrogen-producing Thermoanaerobacterium thermosaccharolyticum SP-H2, resistant to low pH, (2) setting a low initial pH (5.5) to reduce the activity of hydrogenotrophic methanogens, (3) supplementing additives in the form of soluble iron (II) and granular activated carbon (GAC) to stimulate hydrogenase activity and direct interspecies electron transfer (DIET), respectively. For comparison, two methanogenic inoculums with different dominant microbial groups and initial iron concentrations were used. With the simultaneous addition of GAC and iron (II), sequential production of hydrogen and methane was observed for both methanogenic inoculums. Without the addition of GAC, methane formation was practically not observed, which, apparently, indicated the impossibility of syntrophic degradation of acidogenesis products due to the high partial pressure of hydrogen and the absence of conductive material for activation of DIET. Significant differences in hydrogenase activity and the kinetics of hydrogen and methane formation according to the modified Gompertz equation depending on the iron content in the system may indicate the importance of this additive. Further studies should be aimed at determining the optimal initial pH, concentrations of stimulating additives, bioaugmentation and methanogenic cultures, as well as confirming the stability of the proposed single-reactor two-stage anaerobic digestion system in semi-continuous mode.

Effect of different concentrations of sodium selenite on anaerobic digestion of waste sewage sludge

The effect of sodium selenite on anaerobic digestion of waste sewage sludge (WSS) was studied. Interestingly, methane production was enhanced at 2 mg/L and inhibited at 100 mg/L of sodium selenite during the anaerobic digestion of WSS. Hydrolytic activity increased in correlation with increased concentrations of sodium selenite (0 to 100 mg/L). At the acidogenesis stage, acetic acid accumulated at the concentrations more than 25 mg/L of sodium selenite whereas it was consumed for 10 days at the concentration of 2 mg/L of sodium selenite. Also, the concentration of propionic acid decreased and that of butyric acid increased at 100 mg/L of sodium selenite. Although methanogenic activities were totally inhibited at 100 mg/L of sodium selenite, a high hydrogenotrophic methanogenic activity was observed at 2 mg/L of sodium selenite. The abundance of hydrogenotrophic methanogens ( Methanospirillum and Methanocorpusculum ) increased in the WSS at 2 mg/L of sodium selenite. Moreover, the results of pure-culture experiments using Methanosarcina acetivorans C2A show that the growth promotion or inhibition was observed at 2 mg/L or at 100 mg/L of sodium selenite. In conclusion, sodium selenite has the double-sided effect during the anaerobic digestion of WSS as shown by the results that methane production was promoted at a low concentration and inhibited at a high concentration. • Sodium selenite affects methane production through the anaerobic digestion of sewage sludge. • Methanogenic activities were totally inhibited at 100 mg/L of sodium selenite. • Sodium selenite at 2 mg/L enhanced the activity of hydrogenotrophic methanogenesis. • The growth of Methanosarcina acetivorans C2A was promoted at 2 mg/L and inhibited at 100 mg/L of sodium selenite.

Long-term evaluation of the effect of peracetic acid solution on anaerobic wastewater treatment: Process performance and microbial community structure

Peracetic acid (PAA) has been extensively used in poultry processing as an effective disinfectant. However, PAA-bearing poultry processing wastewater can lead to severe upsets of biological wastewater treatment processes. The objective of this study was to assess the long-term effect of PAA solution (PAA and hydrogen peroxide, H2O2) on the anaerobic treatment of poultry processing wastewater. Five reactors were operated for 177 d, semi-continuously fed with poultry processing wastewater and gradually increased levels of H2O2 or PAA solution, added either to the wastewater before feeding (indirect addition), or to the reactors immediately after feeding (direct addition). Indirect, well pre-decomposed H2O2 and PAA had limited effect on biogas production and COD destruction. Biogas production was completely inhibited with direct addition of H2O2 at 227 mg/L, but recovered in ca. 2 d. Direct addition of PAA solution at 500/100 mg/L PAA/H2O2 completely inhibited biogas production and required ca. 18 d incubation for recovery. Methanogenesis was more impacted than fermentation. Long-term H2O2 or PAA solution addition led to a relatively low shift in both bacterial and archaeal communities. The major methanogenic orders were Methanosarcinales and Methanomicrobiales. Direct PAA solution addition decreased both acetoclastic and hydrogenotrophic methanogenic activity; acetoclastic methanogenesis was affected the most.

Zero-valent iron mediated alleviation of methanogenesis inhibition induced by organoarsenic roxarsone

Zero-valent iron (ZVI) can enhance anaerobic digestion, and has great potential to alleviate/eliminate methanogenesis inhibition. Little is known about the feasibility of utilizing ZVI to alleviate methanogenesis inhibition that is caused by typical animal feed additive roxarsone in livestock wastewater. In this study, the role of ZVI on alleviating roxarsone-induced methanogenic inhibition and its mechanisms were investigated. With the increase of roxarsone concentration from 5 to 50 mg/L, the inhibition of methanogenesis increased from 3.0% to 65.7%. This inhibition was alleviated by 80.7% and 57.2% when 1.0 and 10.0 g/L ZVI were added, respectively. Due to ZVI addition, an efficient arsenic immobilization onto ZVI (45.4–85.8%) was achieved mainly through the formation of FeAsO4 precipitate and adsorption by ZVI. Under the function of ZVI, hydrogenotrophic methanogenic activity was obviously restored. The microbial community analysis indicates that the ZVI-regulated alleviation on the methanogenesis inhibition was attributed to the enrichment of Methanobacterium and Methanosarcina. The findings from this study demonstrate that ZVI addition is an effective way for treatment of organoarsenic-contaminated wastewater.

Coupling granular activated carbon and exogenous hydrogen to enhance anaerobic digestion of phenol via predominant syntrophic acetate oxidation and hydrogenotrophic methanogenesis pathway

Anaerobic digestion is a promising biological method for treating phenol-containing wastewater. However, the low methane yield of phenol due to the biological toxicity limits its potential application. This study presents a novel method to enhance the conversion rate of phenol to methane by coupling of granular activated carbon and exogenous hydrogen (GAC/H2). The cumulative methane production in the GAC/H2, H2, GAC, and control groups were 408.2 ± 16.2, 336.5 ± 5.7, 287.2 ± 26. 2, and 258.1 ± 8.6 mL‬ CH4/g COD, respectively. Compared with the control group, the hydrogenotrophic methanogenic activity and electron transfer activity of GAC/H2 group were increased by 403.9 and 367.4%, respectively. The results of the 16SrRNA analysis indicated GAC enhanced the relative abundances of Syntrophus and Syntrophorhabdus, and hydrogen promoted the relative abundances of Cryptanaerobacter, Aminicenantes, and Methanobacterium. Therefore, the coupling of GAC and exogenous hydrogen promoted a dominate SAO-HM pathway to convert phenol to methane.

Ecogenomics Reveals Microbial Metabolic Networks in a Psychrophilic Methanogenic Bioreactor Treating Soy Sauce Production Wastewater.

An ecogenomic analysis of the methanogenic microbial community in a laboratory-scale up-flow anaerobic sludge blanket (UASB) reactor treating soy sauce-processing wastewater revealed a synergistic metabolic network. Granular sludge samples were collected from the UASB reactor operated under psychrophilic (20°C) conditions with a COD removal rate >75%. A 16S rRNA gene amplicon sequencing-based microbial community analysis classified the major microbial taxa as Methanothrix, Methanobacterium, Pelotomaculaceae, Syntrophomonadaceae, Solidesulfovibrio, and members of the phyla Synergistota and Bacteroidota. Draft genomes of dominant microbial populations were recovered by metagenomic shotgun sequencing. Metagenomic- and metatranscriptomic-assisted metabolic reconstructions indicated that Synergistota- and Bacteroidota-related organisms play major roles in the degradation of amino acids. A metagenomic bin of the uncultured Bacteroidales 4484-276 clade encodes genes for proteins that may function in the catabolism of phenylalanine and tyrosine under microaerobic conditions. Syntrophomonadaceae and Pelotomaculaceae oxidize fatty acid byproducts presumably derived from the degradation of amino acids in syntrophic association with aceticlastic and hydrogenotrophic methanogen populations. Solidesulfovibrio organisms are responsible for the reduction of sulfite and may support the activity of hydrogenotrophic methanogens and other microbial populations by providing hydrogen and ammonia using nitrogen fixation-related proteins. Overall, functionally diverse anaerobic organisms unite to form a metabolic network that performs the complete degradation of amino acids in the psychrophilic methanogenic microbiota.

Responses of anaerobic digestion of food waste to coupling effects of inoculum origins, organic loads and pH control under high load: Process performance and microbial characteristics

This study investigated responses of anaerobic digestion (AD) of food waste (FW) with different inocula to varying organic loads and to pH control under high load in terms of process performance and microbial characteristics. Without pH control, digester inoculated by thickened sludge obtained high methane yield of 547.8 ± 27.8 mL/g VS under organic load of 7.5 g VS/L but was inhibited by volatile fatty acids (VFAs) under higher loads (15 and 30 g VS/L). However, digesters inoculated by anaerobic sludge obtained high methane yields of 575.9 ± 34.2, 569.3 ± 24.8 and 531.9 ± 26.2 mL/g VS under organic loads of 7.5, 15 and 30 g VS/L and VFAs inhibition only appeared under extremely high load of 45 g VS/L. Digesters under VFA inhibition with high load were significantly enhanced by controlling single ecological factor pH at 6.5, 7.0 and 7.5, as indicated by shorter lag phases, higher peak values of methane production rate, greater methane yields and fast VFAs degradation. Maximum methane recovery was obtained with pH control at 7.5 under high load. VFA inhibition was accompanied by the degeneration of ecological functions of Syntrophomonadaceae and unidentified Bacteroidales and the dominant growth of unidentified Clostridiales. Under high load and pH control, high stability was strongly associated with obvious growth of Methanosarcina, which enriched methanogenic pathways thus improved system robustness and tolerance to VFAs. Moreover, pH control stimulated the growth of syntrophic Bacteria Syntrophomonadaceae while maintaining the high activity of hydrogenotrophic methanogens therefore sustained efficient syntrophic communities of Bacteria and methanogens and avoided over accumulation of VFAs. pH control promoted adaptive selection of methanogens, leading to obvious decline of archaeal community diversity. This study provided practical guidance on digester configurations of high-load AD of FW and expanded the understanding of responses to coupling effects of inoculum origins, organic loads and pH control under high load concerning process performance and microbial community dynamics.

Blackwater biomethane recovery using a thermophilic upflow anaerobic sludge blanket reactor: Impacts of effluent recirculation on reactor performance

Thermophilic anaerobic digestion is a promising process for high-solid blackwater (BW) treatment due to improved hydrolysis rates, high methanogenesis efficiency, and pathogen removal, when compared with mesophilic treatment. In the present work, the effects of effluent recirculation (i.e., mixing) on thermophilic blackwater treatment were studied. A laboratory-scale thermophilic upflow anaerobic sludge blanket reactor was operated with and without effluent recirculation. The methanogenesis efficiency of the BW treatment increased from 45.0 ± 2.9% when effluent recirculation was applied to 56.7 ± 5.5% without effluent recirculation. Without effluent recirculation, the COD accumulation in the bioreactor was reduced from 17.2 to 3.8% and the effluent volatile fatty acids (VFA) concentration was reduced from 0.64 ± 0.18 to 0.15 ± 0.10 g/L. Further, both acetoclastic and hydrogenotrophic methanogenic activity increased from 101.3 ± 10.8 and 93.9 ± 6.1 to 120.4 ± 9.4 and 118.2 ± 13.2 mg CH4-COD/(gVSS⋅d), respectively, after effluent recirculation was discontinued. The predominant methanogens changed from Methanothermobacter (67%) with effluent recirculation to Methanosarcina (62%) without effluent recirculation. As compared to the effluent recirculation conditions, the enhanced biomethane recovery and treatment performance without effluent recirculation can be attributed to the close proximity of bacteria and archaea groups and the reduced VFA accumulation. Predicted functional gene comparison showed higher prevalence of function for intermediate metabolite transportation (transporters, ATP-binding cassette (ABC) transporters, and two-component system) after discontinuing effluent circulation, which contributed to improved syntrophic propionate oxidation and syntrophic acetate oxidization and enhanced hydrogenotrophic methanogenesis.

Biomethane recovery from source-diverted household blackwater: Impacts from feed sulfate

Biomethane recovery from source-diverted blackwater through anaerobic digestion (AD) offers a sustainable alternative for modern wastewater treatment. Water-wasting conventional toilets consume great amounts of flushing water (9L per flush), which results in blackwater with chemical oxygen demand (COD) of 1006 (±61) mg/L and COD/sulfate ratio of 12.2 (±0.9). In this study, the conventional toilets collected blackwater was treated through an up-flow anaerobic sludge blanket (UASB) reactor at 35°C with a hydraulic retention time (HRT) of 2.2 days, which achieved a COD removal efficiency >80 %. Inhibition in blackwater methanogenesis was observed, which was found associated with the growth of hydrogen utilizing sulfate reducing bacteria (SRB) that competed with hydrogenotrophic methanogens and suppressed the biomethane recovery efficiency. The sludge specific hydrogenotrophic methanogenic activity (SMA [H2&CO2]) increased from 0.37 (±0.02) g CH4-COD/g volatile suspended solids (VSS)/d (treating high sulfate blackwater) to 0.52 (±0.00) g CH4-COD/g VSS/d (treating low sulfate blackwater) when sulfate-free toilet flushing water was adopted (resulting COD/sulfate ratio of 42.9 [±5.0]). This study underlines the importance of considering the impact of sulfate on blackwater methane production when designing future blackwater treatment processes.

Improving the biomethane yield from food waste by boosting hydrogenotrophic methanogenesis

Anaerobic digestion of food waste is usually impacted by high levels of VFAs, resulting in low pH and inhibited methane production from acetate (acetoclastic methanogenesis); however, this could be harnessed for improving methane production via hydrogenotrophic methanogenesis (biomethanation). In this study, batch anaerobic digestion of food waste was conducted to enhance biomethanation by supplying hydrogen gas (H2), using a gas mixture of 5%-H2 and 95%-N2. The addition of H2 influenced a temporal microbial shift in substrate utilisation from dissolved organic nutrients to H2 and CO2 and was perceived to have enhanced the hydrogenotrophic methanogenic activity. As a result, with the release of hydrogen as degradation progressed (secondary fermentation) hydrogenotrophic methanogenesis was further enriched. This resulted in an enhancement of the upgrading of the biogas, with a 12.1% increase in biomethane (from 417.6 to 468.3 NmL-CH4/gVSadded) and 38.9% reduction in CO2 (from 227.1 to 138.7 NmL-CO2/gVSadded). Furthermore, the availability of hydrogen gas at the start of the process promoted faster propionate degradation, by the enhanced activity of the H2-utilisers, thereby, reducing likely propionate-induced inhibitions. The high level of acidification from VFAs production helped to prevent excessive pH increases from the enhanced hydrogenotrophic methanogenic activity. Therefore, it was found that the addition of hydrogen gas to AD reactors treating food waste showed great potential for enhanced methane yield and biogas upgrade, supported by VFAs-induced pH buffer. This creates the possibility to optimise hydrogenotrophic methanogenesis towards obtaining biogas of the right quality for injection into the gas grid.

Optimal the ex-situ biogas biological upgrading to biomethane and its combined application with the anaerobic digestion stage

ABSTRACT Biomethane upgraded from biogas had been proven as a potential renewable energy. Utilizing the metabolic activity of enriched hydrogenotrophic methanogens was regarded as a new way to upgrade biogas. The optimal reaction pH and mixed gas (H2 and CO2) injection time for enriching hydrogenotrophic methanogens were identified, and the performance of upgrading raw biogas produced from food wastes anaerobic digestion was assessed in this study. In the optimal experiment, the results showed that the maximum methane content reached 61.01% when pH was 8 and injection time was 5 min. In all groups, the concentration of acetic acid showed a trend of increasing followed by decreasing during this process. High-throughput sequencing indicated that Methanobacterium and Methanoculleus belonging to hydrogenotrophic methanogens were the dominated genus. The maximum methane content reached 91.43% in the biogas upgrading stage when it was combined with anaerobic digestion stage. At the end of the experiment, the methane content of the upgrading stage was 36.66% higher than that of the raw biogas produced from anaerobic digestion stage. Microbiological analysis showed that hydrogenotrophic methanogens were also the predominant genus.

Enhanced methanogenic performance and metabolic pathway of high solid anaerobic digestion of chicken manure by Fe2+ and Ni2+ supplementation

High solid anaerobic digestion (AD) is a promising technology for the treatment of organic waste in terms of efficient bioenergy production and digestate volume reduction. However, the high solid AD of chicken manure has been a challenge due to the inhibition effects caused by high ammonia levels. In this study, the addition of 280 mg/L Fe2+ and 2 mg/L Ni2+ has been proven to enhance the microbial activity and to overcome inhibition under the stressed environment. The mesophilic digesters feeding with total solid around 150 g/kg operated at an organic loading rate of 4.8 g/(L d) based on volatile solid and a 20-day hydraulic retention time. The total ammonia-nitrogen reached around 6.8 g/L in control and Fe-Ni digesters. The results from the 147-day continuously-fed experiment provided an increment of 34% methane production and a 29% reduction of volatile fatty acids against control. The acetoclastic and hydrogenotrophic methanogenic activity was increased by 89% and 40% respectively. The high throughout sequencing results showed an increased percentage of Methanosarcina sp., which may have contributed to the shifting of the methanogenic pathway towards acetoclastic methanogenesis. The positive effects of Fe2+ and Ni2+ supplementation obtained in this study lay the foundation for its use in AD of nitrogen rich materials.

Adaptation of anaerobic culture to bioconversion of carbon dioxide with hydrogen to biomethane

Biogas is composed of energetically useable methane (CH4) ranging from 50 to 70% and carbon dioxide (CO2) taking up 30–50%. To upgrading biogas to biomethane for better exploitation of its energetical potential, many different physico-chemical processes are commercially applied. Besides these methods, biological methods for the conversion of CO2 with electrolytic H2 to biomethane based on the activity of hydrogenotrophic methanogens are also currently being researched. This study deals with the adaptation of anaerobic culture to H2 and CO2 as substrates under thermophilic and mesophilic conditions. Thermophilic cultures initially showed slower adaptation, but CO2 conversion efficiency was finally by 12.7% higher compared to the mesophilic culture under the same conditions of the H2 loading rate. Thermophilic conditions were favourable also for homoacetogens producing higher concentration of acetate from CO2 and H2 under thermophilic condition compared to mesophilic condition in the excess of H2. Maximum capacity of the used thermophilic suspended biomass for bioconversion of CO2 was the H2 loading rate 2 L/(L.d) resulting in CO2 bioconversion efficiency of 84% (median) in contrast with 40% (median) for mesophilic culture. The system was limited by the concentration of biomass round 4 g/L.

Influence of mesophilic and thermophilic conditions on the anaerobic digestion of food waste: Focus on the microbial activity and removal of long chain fatty acids.

The mesophilic reactor (MR) exhibited advantages in biogas production and performance stability over thermophilic reactor (TR) during the long-term anaerobic digestion (AD) of food waste (FW) with stepwise organic loading rate elevating. It was interesting to explore the mechanism causing the divergences in performances between these two reactors. The microbial activity was compared on day 110 when TR began to deteriorate. The results show that MR had significantly higher specific acetoclastic methanogenic activities (SAMA) and specific propionate and butyrate oxidative activities (SPOA and SBOA) than TR. The SAMA, SPOA and SBOA in TR were only 50.3%, 18.6% and 46.4% of those values in MR, respectively. Remarkably, the specific hydrogenotrophic methanogenic activity of 15.5±2.1, 15.7±4.6 mmol CH4·L-1 original slurry·d-1 in MR and TR was comparative with insignificant difference, which indicates that the microbial activity in TR had been inhibited widely apart from the hydrogenotrophic methanogenesis. Additionally, many particles with the diameters of 1-2 mm were observed to form in MR and identified as complexes of calcium and long chain fatty acids (LCFAs). The formation of calcium crystallization might alleviate the inhibition of LCFAs during AD of FW, which further supports the better performance in MR than TR.

Acclimation to extremely high ammonia levels in continuous biomethanation process and the associated microbial community dynamics

Acclimatized anaerobic communities to high ammonia levels can offer a solution to the ammonia toxicity problem in biogas reactors. In the current study, a stepwise acclimation strategy up to 10g NH4+-N L−1, was performed in mesophilic (37±1°C) continuously stirred tank reactors. The reactors were co-digesting (20/80 based on volatile solid) cattle slurry and microalgae, a protein-rich, 3rd generation biomass. Throughout the acclimation period, methane production was stable with more than 95% of the uninhibited yield. Next generation 16S rRNA gene sequencing revealed a dramatic microbiome change throughout the ammonia acclimation process. Clostridium ultunense, a syntrophic acetate oxidizing bacteria, increased significantly alongside with hydrogenotrophic methanogen Methanoculleus spp., indicating strong hydrogenotrophic methanogenic activity at extreme ammonia levels (>7g NH4+-N L−1). Overall, this study demonstrated for the first time that acclimation of methanogenic communities to extreme ammonia levels in continuous AD process is possible, by developing a specialised acclimation AD microbiome.