Methane yield response to pretreatment is dependent on substrate chemical composition: a meta-analysis on anaerobic digestion systems.

Proper pretreatment of organic residues prior to anaerobic digestion (AD) can maximize global biogas production from varying sources without increasing the amount of digestate, contributing to global decarbonization goals. However, the efficiency of pretreatments applied on varying organic streams is poorly assessed. Thus, we performed a meta-analysis on AD studies to evaluate the efficiencies of pretreatments with respect to biogas production measured as methane yield. Based on 1374 observations our analysis shows that pretreatment efficiency is dependent on substrate chemical dominance. Grouping substrates by chemical composition e.g., lignocellulosic-, protein- and lipid-rich dominance helps to highlight the appropriate choice of pretreatment that supports maximum substrate degradation and more efficient conversion to biogas. Methane yield can undergo an impactful increase compared to untreated controls if proper pretreatment of substrates of a given chemical dominance is applied. Non-significant or even adverse effects on AD are, however, observed when the substrate chemical dominance is disregarded.

Deciphering the contribution of microbial biomass to the properties of dissolved and particulate organic matter in anaerobic digestates.

The recalcitrant structures either from substrate or microbial biomass contained in digestates after anaerobic digestion (AD) highly influence digestate valorization. To properly assess the microbial biomass contribution to the digested organic matter (OM), a combination of characterization methods and the use of various substrate types in anaerobic continuous reactors was required. The use of totally biodegradable substrates allowed detecting soluble microbial products via fluorescence spectroscopy at emission wavelengths of 420 and 460 nm while the protein-like signature was enhanced by the whey protein. During reactors' operation, a transfer of complex compounds to the dissolved OM from the particulate OM was observed through fluorescence applied on biochemical fractionation. Consequently, the fluorescence complexity index of the dissolved OM increased from 0.59-0.60 to 1.06-1.07, whereas it decreased inversely for the extractable soluble from the particulate OM from 1.16-1.19 to 0.42-0.54. Accordingly, fluorescence regional integration showed differences among reactors based on visual inspection and orthogonal partial latent structures (OPLS) analysis. Similarly, the impact of the substrate type and operation time on the particulate OM was revealed by 13C nuclear magnetic resonance using OPLS, providing a good model (R2X = 0.93 and Q2 = 0.8) with a clear time-trend. A high signal resonated at ∼30 ppm attributed to CH2-groups in the aliphatic chain of lipid-like structure besides carbohydrates intensities at 60-110 ppm distinguished the reactor fed with whey protein from the other, which was mostly biomass related. Indeed, this latter displayed a higher presence of peptidoglycan (δH/C: 1.6-2.0/20-25 ppm) derived from microbial biomass by 1H-13C heteronuclear single-quantum coherence (HSQC) nuclear magnetic resonance. Interestingly, the sample distribution obtained by non-metric multidimensional scaling of bacterial communities resembled the attained using 13C NMR properties, opening new research perspectives. Overall, this study discloses the microbial biomass contribution to digestates composition to improve the OM transformation mechanism knowledge.

Absence of oxygen effect on microbial structure and methane production during drying and rewetting events.

Natural environments with frequent drainage experience drying and rewetting events that impose fluctuations in water availability and oxygen exposure. These relatively dramatic cycles profoundly impact microbial activity in the environment and subsequent emissions of methane and carbon dioxide. In this study, we mimicked drying and rewetting events by submitting methanogenic communities from strictly anaerobic environments (anaerobic digestors) with different phylogenetic structures to consecutive desiccation events under aerobic (air) and anaerobic (nitrogen) conditions followed by rewetting. We showed that methane production quickly recovered after each rewetting, and surprisingly, no significant difference was observed between the effects of the aerobic or anaerobic desiccation events. There was a slight change in the microbial community structure and a decrease in methane production rates after consecutive drying and rewetting, which can be attributed to a depletion of the pool of available organic matter or the inhibition of the methanogenic communities. These observations indicate that in comparison to the drying and rewetting events or oxygen exposure, the initial phylogenetic structure and the organic matter quantity and quality exhibited a stronger influence on the methanogenic communities and overall microbial community responses. These results change the current paradigm of the sensitivity of strict anaerobic microorganisms to oxygen exposure.

Identifying targets for increased biogas production through chemical and organic matter characterization of digestate from full-scale biogas plants: what remains and why?

BACKGROUND: This study examines the destiny of macromolecules in different full-scale biogas processes. From previous studies it is clear that the residual organic matter in outgoing digestates can have significant biogas potential, but the factors dictating the size and composition of this residual fraction and how they correlate with the residual methane potential (RMP) are not fully understood. The aim of this study was to generate additional knowledge of the composition of residual digestate fractions and to understand how they correlate with various operational and chemical parameters. The organic composition of both the substrates and digestates from nine biogas plants operating on food waste, sewage sludge, or agricultural waste was characterized and the residual organic fractions were linked to substrate type, trace metal content, ammonia concentration, operational parameters, RMP, and enzyme activity. RESULTS: Carbohydrates represented the largest fraction of the total VS (32-68%) in most substrates. However, in the digestates protein was instead the most abundant residual macromolecule in almost all plants (3-21 g/kg). The degradation efficiency of proteins generally lower (28-79%) compared to carbohydrates (67-94%) and fats (86-91%). High residual protein content was coupled to recalcitrant protein fractions and microbial biomass, either from the substrate or formed in the degradation process. Co-digesting sewage sludge with fat increased the protein degradation efficiency with 18%, possibly through a priming mechanism where addition of easily degradable substrates also triggers the degradation of more complex fractions. In this study, high residual methane production (> 140 L CH4/kg VS) was firstly coupled to operation at unstable process conditions caused mainly by ammonia inhibition (0.74 mg NH3-N/kg) and/or trace element deficiency and, secondly, to short hydraulic retention time (HRT) (55 days) relative to the slow digestion of agricultural waste and manure. CONCLUSIONS: Operation at unstable conditions was one reason for the high residual macromolecule content and high RMP. The outgoing protein content was relatively high in all digesters and improving the degradation of proteins represents one important way to increase the VS reduction and methane production in biogas plants. Post-treatment or post-digestion of digestates, targeting microbial biomass or recalcitrant protein fractions, is a potential way to achieve increased protein degradation.

Effect of Cobalt, Nickel, and Selenium/Tungsten Deficiency on Mesophilic Anaerobic Digestion of Chemically Defined Soluble Organic Compounds.

Trace elements (TEs) are vital for anaerobic digestion (AD), due to their role as cofactors in many key enzymes. The aim of this study was to evaluate the effects of specific TE deficiencies on mixed microbial communities during AD of soluble polymer-free substrates, thus focusing on AD after hydrolysis. Three mesophilic (37 °C) continuous stirred-tank biogas reactors were depleted either of Co, Ni, or a combination of Se and W, respectively, by discontinuing their supplementation. Ni and Se/W depletion led to changes in methane kinetics, linked to progressive volatile fatty acid (VFA) accumulation, eventually resulting in process failure. No significant changes occurred in the Co-depleted reactor, indicating that the amount of Co present in the substrate in absence of supplementation was sufficient to maintain process stability. Archaeal communities remained fairly stable independent of TE concentrations, while bacterial communities gradually changed with VFA accumulation in Ni- and Se-/W-depleted reactors. Despite this, the communities remained relatively similar between these two reactors, suggesting that the major shifts in composition likely occurred due to the accumulating VFAs. Overall, the results indicate that Ni and Se/W depletion primarily lead to slower metabolic activities of methanogenic archaea and their syntrophic partners, which then has a ripple effect throughout the microbial community due to a gradual accumulation of intermediate fermentation products.

Substrate and operational conditions as regulators of fluid properties in full-scale continuous stirred-tank biogas reactors - implications for rheology-driven power requirements.

Understanding fluid rheology is important for optimal design and operation of continuous stirred-tank biogas reactors (CSTBRs) and is the basis for power requirement estimates. Conflicting results have been reported regarding the applicability of total solid (TS) and/or total volatile solid (TVS) contents of CSTBR fluids as proxies for rheological properties. Thus, the present study investigates relationships between rheological properties of 12 full-scale CSTBR fluids, their substrate profiles, and major operational conditions, including pH, TS and TVS contents, organic loading rate, hydraulic retention time, and temperature. Rheology-driven power requirements based on various fluid characteristics were evaluated for a general biogas reactor setup. The results revealed a significant correlation only between the rheological fluid properties and TS or TVS contents for sewage sludge digesters and thermophilic co-digesters (CD), but not for mesophilic CD. Furthermore, the calculated power requirements for pumping and mixing, based on the various fluid characteristics of the studied CSTBRs, varied broadly irrespective of TS and TVS contents. Thus, this study shows that the TS and/or TVS contents of digester fluid are not reliable estimators of the rheological properties in CSTBRs digesting substrates other than sewage sludge.

Microbial rRNA gene expression and co-occurrence profiles associate with biokinetics and elemental composition in full-scale anaerobic digesters.

This study examined whether the abundance and expression of microbial 16S rRNA genes were associated with elemental concentrations and substrate conversion biokinetics in 20 full-scale anaerobic digesters, including seven municipal sewage sludge (SS) digesters and 13 industrial codigesters. SS digester contents had higher methane production rates from acetate, propionate and phenyl acetate compared to industrial codigesters. SS digesters and industrial codigesters were distinctly clustered based on their elemental concentrations, with higher concentrations of NH3 -N, Cl, K and Na observed in codigesters. Amplicon sequencing of 16S rRNA genes and reverse-transcribed 16S rRNA revealed divergent grouping of microbial communities between mesophilic SS digesters, mesophilic codigesters and thermophilic digesters. Higher intradigester distances between Archaea 16S rRNA and rRNA gene profiles were observed in mesophilic codigesters, which also had the lowest acetate utilization biokinetics. Constrained ordination showed that microbial rRNA and rRNA gene profiles were significantly associated with maximum methane production rates from acetate, propionate, oleate and phenyl acetate, as well as concentrations of NH3 -N, Fe, S, Mo and Ni. A co-occurrence network of rRNA gene expression confirmed the three main clusters of anaerobic digester communities based on active populations. Syntrophic and methanogenic taxa were highly represented within the subnetworks, indicating that obligate energy-sharing partnerships play critical roles in stabilizing the digester microbiome. Overall, these results provide new evidence showing that different feed substrates associate with different micronutrient compositions in anaerobic digesters, which in turn may influence microbial abundance, activity and function.

Chemical speciation of sulfur and metals in biogas reactors - Implications for cobalt and nickel bio-uptake processes.

This article deals with the interrelationship between overall chemical speciation of S, Fe, Co, and Ni in relation to metals bio-uptake processes in continuous stirred tank biogas reactors (CSTBR). To address this topic, laboratory CSTBRs digesting sulfur(S)-rich stillage, as well as full-scale CSTBRs treating sewage sludge and various combinations of organic wastes, termed co-digestion, were targeted. Sulfur speciation was evaluated using acid volatile sulfide extraction and X-ray absorption spectroscopy. Metal speciation was evaluated by chemical fractionation, kinetic and thermodynamic analyses. Relative Fe to S content is identified as a critical factor for chemical speciation and bio-uptake of metals. In reactors treating sewage sludge, quantity of Fe exceeds that of S, inducing Fe-dominated conditions, while sulfide dominates in laboratory and co-digestion reactors due to an excess of S over Fe. Under sulfide-dominated conditions, metals availability for microorganisms is restricted due to formation of metal-sulfide precipitates. However, aqueous concentrations of different Co and Ni species were shown to be sufficient to support metal acquisition by microorganisms under sulfidic conditions. Concentrations of free metal ions and labile metal complexes in aqueous phase, which directly participate in bio-uptake processes, are higher under Fe-dominated conditions. This in turn enhances metal adsorption on cell surfaces and bio-uptake rates.

Microbial community adaptation influences long-chain fatty acid conversion during anaerobic codigestion of fats, oils, and grease with municipal sludge.

Codigesting fats, oils, and greases with municipal wastewater sludge can greatly improve biomethane recovery at wastewater treatment facilities. Process loading rates of fats, oils, and greases have been previously tested with little knowledge of the digester microbial community structure, and high transient fat loadings have led to long chain fatty acid (LCFA) accumulation and digester upsets. This study utilized recently-developed quantitative PCR assays for syntrophic LCFA-degrading bacteria along with 16S amplicon sequencing to relate changes in microbial community structure to LCFA accumulation during transient loading increases to an anaerobic codigester receiving waste restaurant oil and municipal wastewater sludge. The 16S rRNA gene concentration of the syntrophic ß-oxidizing genus Syntrophomonas increased to ∼15% of the Bacteria community in the codigester, but stayed below 3% in the control digester that was fed only wastewater sludge. Methanosaeta and Methanospirillum were the dominant methanogenic genera enriched in the codigester, and together comprised over 80% of the Archaea community by the end of the experimental period. Constrained ordination showed that changes in the codigester Bacteria and Archaea community structures were related to measures of digester performance. Notably, the effluent LCFA concentration in the codigester was positively correlated to the specific loading rate of waste oil normalized to the Syntrophomonas 16S rRNA concentration. Specific loading rates of 0-1.5 × 10(-12) g VS oil/16S gene copies-day resulted in LCFA concentrations below 30 mg/g TS, whereas LCFA accumulated up to 104 mg/g TS at higher transient loading rates. Based on the community-dependent loading limitations found, enhanced biomethane production from high loadings of fats, oils and greases can be achieved by promoting a higher biomass of slow-growing syntrophic consortia, such as with longer digester solids retention times. This work also demonstrates the potential for controlling the loading rate of fats, oils, and greases based on the analysis of the codigester community structure, such as with quantitative PCR measurements of syntrophic LCFA-degrading bacteria abundance.

Geochemical control processes and potential sediment toxicity in a mine-impacted lake.

Geochemical parameters and major ion concentrations from sediments of a freshwater lake in the town of Åtvidaberg, southeastern, Sweden, were used to identify the geochemical processes that control the water chemistry. The lake sediments are anoxic, characterized by reduced sulfur and sulfidic minerals. The hypothesis tested is that in sulfidic-anaerobic contaminated sediments, the presence of redox potential changes creates a favorable condition for sulfide oxidation, resulting in the release of potentially toxic metals. The acid volatile sulfide (AVS) contents ranged from 5.5 µmol/g to 16 µmol/g of dry sediment. Comparison of total mine tailing metals (∑mine tailing metals) with simultaneously extracted metals (SEM) in sediments indicates that up to 20% of the ∑mine tailing metals are bound to the solid phase as AVS. Consequently, the AVS and SEM analysis classified all sediment samples as potentially toxic in terms of heavy metal concentrations (i.e., SEM to AVS ratio distribution > 1). Evaluation of hydrogeochemical data suggests that calcite dissolution, iron (III) oxyhydroxysulfate mineral jarosite (H-jarosite) precipitation, hematite precipitation, and siderite precipitation are the most prevailing geochemical processes that control the geochemical interactions between the water column and sediment in a mine-impacted lake. The geochemical processes were verified and quantified using a chemical equilibrium modeling program, Visual MINTEQ, Ver 3.1, beta. The identified geochemical processes create an environment in which the characteristics of sulfate-rich waters and acidic-iron produce the geochemical conditions for acid mine drainage and mobilization of toxic metals.

Characterization of dissolved organic matter in full scale continuous stirred tank biogas reactors using ultrahigh resolution mass spectrometry: a qualitative overview.

Dissolved organic matter (DOM) was characterized in eight full scale continuous stirred tank biogas reactors (CSTBR) using solid-phase extraction and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FT-ICR-MS). An overview of the DOM molecular complexity in the samples from biogas reactors with conventional operational conditions and various substrate profiles is provided by assignments of unambiguous exact molecular formulas for each measured mass peak. Analysis of triplicate samples for each reactor demonstrated the reproducibility of the solid-phase extraction procedure and ESI-FT-ICR-MS which allowed precise evaluation of the DOM molecular differences among the different reactors. Cluster analysis on mass spectrometric data set showed that the biogas reactors treating sewage sludge had distinctly different DOM characteristics compared to the codigesters treating a combination of organic wastes. Furthermore, the samples from thermophilic and mesophilic codigesters had different DOM composition in terms of identified masses and corresponding intensities. Despite the differences, the results demonstrated that compositionally linked organic compounds comprising 28-59% of the total number of assigned formulas for the samples were shared in all the reactors. This suggested that the shared assigned formulas in studied CSTBRs might be related to common biochemical transformation in anaerobic digestion process and therefore, performance of the CSTBRs.