Search Results (340)

The organic loading rate (OLR) is a crucial parameter in the anaerobic digestion of lignocellulosic biomass. Optimizing the OLR ensures a balanced substrate release for gradual hydrolysis, thereby preventing the accumulation of inhibitors that can disrupt methanogenesis. Its significance lies in its direct impact on the stability, efficiency, and overall performance of the digestion process. This study investigated the long-term anaerobic co-digestion of lignocellulosic biomass (Sida hermaphrodita) and cattle manure under varying organic loading rates (S1: 2 kgVS/m³·d, S2: 3 kgVS/m³·d, and S3: 4 kgVS/m³·d). Methane production, microbial community dynamics, and volatile fatty acid (VFA) profiles were analyzed. During S1 and S2, methane production was stable, achieving 446.3 ± 153.7 NL/kgVS and 773.4 ± 107.8 NL/kgVS, respectively. However, at S3, methane production declined, accompanied by a pH drop from 7.68 to 6.11, an increase in the FOS/TAC ratio from 0.272 to 0.35, and the accumulation of acetic and propionic acids at the end of the digestion cycle. Microbial analysis revealed that the abundance of Firmicutes increased with higher OLRs, reaching 93.6% in S3, while the Bacteroidota abundance decreased, reaching 3.0% in S3. During S1, methane production occurred through both acetoclastic and hydrogenotrophic pathways. Full article

The pervasive presence of microplastics (MPs) in agroecosystems poses a significant threat to soil health and plant growth. This study investigates the effects of varying concentrations and sizes of polystyrene microplastics (PS-MPs) on the Solanum lycopersicum L.’s height, dry weight, antioxidant enzyme activities, soil physicochemical properties, and rhizosphere microbial communities. The results showed that the PS0510 treatment significantly increased plant height (93.70 cm, +40.83%) and dry weight (2.98 g, +100%). Additionally, antioxidant enzyme activities improved across treatments for S. lycopersicum L. roots. Physicochemical analyses revealed enhanced soil organic matter and nutrient levels, including ammonium nitrogen, phosphorus, and effective potassium. Using 16S rRNA sequencing and molecular ecological network techniques, we found that PS-MPs altered the structure and function of the rhizosphere microbial community associated with S. lycopersicum L. The PS1005 treatment notably increased microbial diversity and displayed the most complex ecological network, while PS1010 led to reduced network complexity and more negative interactions. Linear discriminant analysis effect size (LEfSe) analysis identified biomarkers at various taxonomic levels, reflecting the impact of PS-MPs on microbial community structure. Mantel tests indicated positive correlations between microbial diversity and soil antioxidant enzyme activity, as well as relationships between soil physicochemical properties and enzyme activity. Predictions of gene function revealed that PS-MP treatments modified carbon and nitrogen cycling pathways, with PS1005 enhancing methanogenesis genes (mcrABG) and PS1010 negatively affecting denitrification genes (nirK, nirS). This study provides evidence of the complex effects of PS-MPs on soil health and agroecosystem functioning, highlighting their potential to alter soil properties and microbial communities, thereby affecting plant growth. Full article

Hydrogen is a pivotal energy carrier for achieving sustainability and stability, but safe and efficient geological underground hydrogen storage (UHS) is critical for its large-scale application. This study investigates the impacts of geochemical and biochemical reactions on UHS, addressing challenges that threaten storage efficiency and safety. Geochemical reactions in saline aquifers, particularly the generation of hydrogen sulfide (H2S), were analyzed using advanced compositional and geochemical modeling calibrated with experimental kinetic data. The results indicate that geochemical reactions have a minimal effect on hydrogen consumption. However, by year 10 of storage operations, H₂S levels could reach 12–13 ppm, necessitating desulfurization to maintain storage performance and safety. The study also examines the methanogenesis reaction, where microorganisms consume hydrogen and carbon dioxide to produce methane. Numerical simulations reveal that microbial activity under suitable conditions can reduce in situ hydrogen volume by up to 50%, presenting a critical hurdle to UHS feasibility. These findings highlight the necessity of conducting microbial analyses of reservoir brines during the screening phase to mitigate hydrogen losses. The novelty of this work lies in its comprehensive field-scale analysis of impurity-induced geochemical and microbial reactions and their implications for underground hydrogen storage. By integrating kinetic parameters derived from experimental data with advanced computational modeling, this study uncovers the mechanisms driving these reactions and highlights their impact on storage efficiency, and safety. By offering a detailed field-scale perspective, the findings provide a pivotal framework for advancing future hydrogen storage projects and ensuring their practical viability. Full article

Microplastics (MPs) are widely distributed in the environment, and they inevitably enter animal bodies during livestock and poultry farming, leading to their presence in livestock and poultry manure. However, there is limited research on the effects of different types of MPs on the anaerobic digestion (AD) performance of livestock and poultry manure. Herein, we investigated the impact of four types of MPs (polyvinyl chloride (PVC), polypropylene (PP), polyethylene (PE), and polyhydroxyalkanoate (PHA)) on AD performance using cattle manure as a substrate. Results demonstrated that the cumulative methane production in the PE group reached 5568.05 mL, exhibiting an 11.97% increase compared to the control group. Conversely, the cumulative methane production was decreased by 5.52%, 9.69%, and 14.48% in the PP, PVC, and PHA groups, respectively. Physicochemical analyses showed that MPs promoted organic matter hydrolysis on day 4 of AD, leading to the accumulation of volatile fatty acids (VFAs) in the initial stage. Specifically, the acetic acid content of PE was 44.48–92.07 mL/L higher than that of the control during the first 8 days. PE MPs also enriched microorganisms associated with methane production. The abundance of Firmicutes was enhanced by 2.89–17.57%, Methanosaeta by 8.42–12.48%, and Methanospirillum by 10.91–16.89% in comparison to the control; whereas PHA MPs decreased the abundance of Methanosaeta by 8.14–31.40%. Moreover, PHA MPs inhibited methane production by suppressing acetate kinase activity while promoting lactate dehydrogenase release from microorganisms involved in the AD process. Based on changes observed in key enzyme functional gene abundances, PHA MPs reduced acetyl-CoA carboxylase functional gene abundance, negatively affecting the acetone cleavage methanogenesis pathway. Meanwhile, PE MPs significantly increased acetate-CoA ligase abundance, thereby promoting the acetic acid methanogenesis pathway. The results provide novel insights into the influence exerted by MPs on AD performance when applied to livestock manure. Full article

This study explores the impact of incorporating conductive materials and bioelectrochemical systems (BES) on the efficiency of anaerobic digestion (AD) of sewage sludge. The research consists of two phases: biodegradability tests using 3D-printed polylactic acid-based conductive fillers (PLA/Carbon Black and PLA/Graphene) and semi-continuous assays integrating an external BES into the AD process. Results from biodegradability tests indicate that conductive fillers enhance around 50% methane production during the start-up phase, with microbial communities adapting over time to reduce variability in methane yields. Moreover, as the experiment progressed, the methane yields of the digesters with and without fillers became equal. Semi-continuous experiments demonstrate that BES integration improves process stability and methane production by achieving a 5–10% improvement in the amount of methane in the biogas throughout the entire operation, even under high organic loads, by facilitating diverse electron transfer pathways. The challenges of BES operation highlight the need for optimized designs to ensure scalability. Microbial analyses reveal that BES application shifts methanogenic pathways, favouring acetoclastic methanogenesis. Overall, the findings underscore the potential of conductive materials and BES to improve biogas quality and production, contributing to sustainable wastewater management and renewable energy generation. Full article

When vegetable waste (VW) is used as a sole substrate for anaerobic digestion (AD), the rapid accumulation of volatile fatty acids (VFAs) can impede interspecies electron transfer (IET), resulting in a relatively low biogas production rate. In this study, Chinese cabbage and cabbage were selected as the VW substrates, and four continuous stirred tank reactors (CSTRs) were employed. Different concentrations of biochar-loaded nano-Fe₃O₄(Fe₃O₄@BC) (100 mg/L, 200 mg/L, 300 mg/L) were added, and the organic loading rate (OLR) was gradually increased during the AD process. The changes in biogas production rate, VFAs, and microbial community structure in the fermentation tanks were analyzed to identify the optimal dosage of Fe₃O₄@BC and the maximum OLR. The results indicated that at the maximum OLR of 3.715 g (VS)/L·d, the addition of 200 mg/L of Fe₃O₄@BC most effectively promoted an increase in the biogas production rate and reduced the accumulation of VFAs compared to the other treatments. Under these conditions, the biogas production rate reached 0.658 L/g (VS). Furthermore, the addition of Fe₃O₄@BC enhanced both the diversity and abundance of bacteria and archaea. At the genus level, the abundance of Christensenellaceae_R-7_group, Sphaerochaeta, and the archaeal genus Thermovirga was notably increased. Full article

Biochar-based additives can enhance the ability to produce methane during anaerobic digestion (AD), and biogas residues (BRs) are solid waste that can cause environmental pollution. Therefore, in this work, BRs were used as raw material to prepare biochar, and Fe³⁺ was used to modify biochar for use in the AD process, generating pig manure water (PMW). The results showed that biogas residue biochar (BRB) showed good pore size and had a “honeycomb structure” on its surface. The commercially available iron–carbon composite material (Fe-C) showed the greatest cumulative methane production (CMP), the greatest removal rates of chemical oxygen demand (COD), and the greatest degradation rate of volatile fatty acids (VFAs), with the order of Fe-C > Fe³⁺-modified biogas residue biochar (FBRB) > BRB > control group. Fe³⁺ and Fe⁰ showed similar effects, where both could enhance the methanogenesis performance of anaerobic digestion by promoting direct interspecific electron transfer, and Fe⁰ was slightly more effective than Fe³⁺. Bacteroidotas and Firmicutes were the predominant phyla, and Clostridium_sensu_stricto_1 was the predominant genus. The addition of biochar and Fe³⁺ promoted the transformation of microorganisms from the conventional metabolic mode into an efficient metabolic mode. Extracellular electron transfer played a crucial role in this. Full article

The impact of macroalgae species on rumen function remains largely unexplored. This present study aimed to identify the biocompounds of the three types of marine macroalgae described: Macrocystis pyrifera (Brown), Ulva spp. (Lettuce), Mazzaella spp. (Red) and their effect on species-specific modulations of the rumen microbiome. The macroalgae were characterized using GC-MS. Twelve Rambouillet lambs were randomly assigned to one of four experimental diets (n = 3 per treatment): (a) control diet (CD); (b) CD + 5 g of Red algae; (c) CD + 5 g of Brown algae; and (d) CD + 5 g of Lettuce algae. After the lambs ended their fattening phase, they donated ruminal fluid for DNA extraction and 16S rRNA gene V3 amplicon sequencing. Results: The tagged 16S rRNA amplicon sequencing and statistical analysis revealed that the dominant ruminal bacteria shared by all four sample groups belonged to phyla Firmicutes and Bacteroidota. However, the relative abundance of these bacterial groups was markedly affected by diet composition. In animals fed with macroalgae, the fibrinolytic and cellulolytic bacteria Selenomonas was found in the highest abundance. The diversity in chemical composition among macroalgae species introduces a range of bioactive compounds, particularly VOCs like anethole, beta-himachalene, and 4-ethylphenol, which demonstrate antimicrobial and fermentation-modulating properties. Full article

Propionate, a critical intermediate in anaerobic digestion, and its syntrophic removal, is sensitive to stress. To our knowledge, this study investigates for the first time the response of a metabolic gene panel to organic loading rate (OLR) stress in propionate-degrading methanogenic consortia in lab-scale upflow anaerobic sludge blanket (UASB) reactors. The experimental phases included stabilisation (1.4–2.8 g COD/L/day), electroactive enrichment, OLR shock (6 g COD/L/day), and early recovery. Quantitative PCR was used to assess the abundance of key functional genes (16SrRNA, mcrA, pilA, and hgtR). During stabilisation, ~200 mLCH₄/h was produced, the mcrA/16SrRNA ratio was 0.78–2.64, and pilA and hgtR abundances were 1.29–2.27 × 10⁵ and 2.12–4.37 × 10⁴ copies/gVS. Following the OLR shock, methane production ceased entirely, accompanied by a sharp decline in the mcrA/16S ratio (0.08–0.24) and significant reductions in pilA (1.43-log) and hgtR (1.34-log) abundance. Partial recovery of pilA and hgtR abundance (1.19 × 10⁵ and 8.57 × 10⁴) was observed in the control reactor after the early recovery phase. The results highlight the utility of mcrA, 16SrRNA, pilA, and associated ratios, as reliable indicators of OLR stress in lab-scale UASB reactors. This study advances the understanding of molecular stress responses in propionate-degrading methanogenic consortia, focusing on direct interspecies electron transfer in process stability and recovery. Full article

The increasing environmental pollution resulting from plastic waste and the need to reuse agro-industrial wastes as a source of discarding has led to the development of innovative biobased products. In the frame of this context, the use of neat polylactic acid (PLA) and its blend with polybutylene succinate (PBS) with or without cellulose nanocrystals (CNCs) extracted from hemp fibers is explored here. This study aimed to assess the biogas production of different biopolymeric films. In parallel, life cycle assessment (LCA) analysis was performed on the same films, focusing on their production phase and potential end-of-life scenarios, regardless of film durability (i.e., single-use packaging) and barrier performance, to counteract possible soil health threats. Specifically, this study considered three specific systems: PLA, PLA_PBS (PLA/PBS blend 80:20 w/w), and PLA_PBS_3CNC (PLA/PBS blend + 3% CNCs) films. The assessment involved a batch anaerobic digestion (AD) process at 52 °C, using digestate obtained from the anaerobic treatment of municipal waste as the inoculum and cellulose as a reference material. The AD process was monitored over about 30 days, revealing that reactors containing cellulose showed inherent biodegradability and enhanced biogas production. On the other hand, biopolymeric films based on PLA and its blends with PBS and CNCs exhibited an inhibitory effect, likely due to their recalcitrant nature, which can limit or delay microbial activity toward biomass degradation and methanogenesis. LCA analysis was performed taking into consideration the complex environmental implications of both including biopolymers in the production of renewable energy and the use of post-composting digestate as an organic fertilizer. Remarkably, the PLA_PBS_3CNC formulation revealed slightly superior performance in terms of biodegradability and biogas production, mainly correlated to the presence of CNCs in the blend. The observed enhanced biodegradability and biogas yield, coupled with the reduced environmental impact, confirm the key role of optimized biopolymeric formulations in mitigating inhibitory effects on AD processes while maximizing, at the same time, the utilization of naturally derived energy sources. Full article

Recent studies actively debate oxic methane (CH₄) production processes in water and terrestrial ecosystems. This previously unknown source of CH₄ on a regional and global scale has the potential to alter our understanding of climate-driving processes in vulnerable ecosystems, particularly high-latitude ecosystems. Thus, the main objective of this study is to use the incubation approach to explore possible greenhouse gas (GHG) fluxes by the most widely distributed species of epiphytic lichens (ELs; Evernia mesomorpha Nyl. and Bryoria simplicior (Vain.) Brodo et D. Hawksw.) in the permafrost zone of Central Siberia. We observed CH₄ production by hydrated (50%–400% of thallus water content) ELs during 2 h incubation under illumination. Moreover, in agreement with other studies, we found evidence that oxic CH₄ production by Els is linked to the CO₂ photoassimilation process, and the EL thallus water content regulates that relationship. Although the GHG fluxes presented here were obtained under a controlled environment and are probably not representative of actual emissions in the field, more research is needed to fully comprehend ELs’ function in the C cycle. This particular research provides a solid foundation for future studies into the role of ELs in the C cycle of permafrost forest ecosystems under ongoing climate change (as non-methanogenesis processes in oxic environments). Full article

Precise control of operational parameters in anaerobic digestion reactors is crucial to avoid imbalances that could affect biomethane production and alterations in the microbiota. Restoring the methanogenic microbiota after a failure is essential for recovering methane production, yet no published strategies exist for this recovery. In this study, we restored the methanogenic microbiota in an anaerobic SBR reactor that operates with both biofilm and suspended biomass simultaneously, aiming to treat tequila vinasses. Four strategies were evaluated for restoring the methanogenic microbiota: reducing the initial vinasse concentration, increasing the reaction time (RT), adjusting the carbon/nitrogen (C/N) ratio, and progressively increasing the initial vinasse concentration. Among these, adjusting the C/N ratio emerged as a critical parameter for restoring organic matter removal efficiency and reestablishing methanogenic microbiota. The operational conditions under which the methanogenic activity and microbiota were restored were as follows: Operating the A-SBR with an initial vinasse concentration of 60%, an RT of 168 h, a pH of 6.9 ± 0.2, a temperature of 35 ± 2 °C, and a C/N ratio adjusted to 100/1.9 resulted in stable COD removal efficiency of 93 ± 3% over a year and a high percentage of methanogenic microorganisms in both the suspended microbiota (69%) and biofilm (52%). The normalized methane production (0.332 NL CH₄/g CODr) approached the theoretical maximum value (0.35 L CH₄/g CODr) after restoring the population and methanogenic activity within the reactor. Full article

Biomethane production via biogas upgrading is regarded as a future renewable gas, further boosting the biogas economy. Moreover, when upgrading is realized by the biogas CO₂ conversion to CH₄ using surplus renewable energy, the process of upgrading becomes a renewable energy storage method. This conversion can be carried out via microorganisms, and has attracted scientific attention, especially under thermophilic conditions. In this study, mesophilic conditions were imposed using a previously developed enriched culture. The enriched culture consisted of the hydrogenotrophic Methanobrevibacter (97% of the Archaea species and 60% of the overall population). Biogas upgrading took place in three lab-scale bioreactors: (a) a 1.2 L bubble reactor (BR), (b) a 2 L trickling bed reactor (TBR) filled with plastic supporting material (TBR-P), and (c) a 1.2 L TBR filled with sintered glass balls (TBR-S). The gas fed into the reactors was a mixture of synthetic biogas and hydrogen, with the H₂ to biogas CO₂ ratio being 3.7:1, lower than the stoichiometric ratio (4:1). Therefore, the feeding gas mixture did not make it possible for the CH₄ content in the biomethane to be more than 97%. The results showed that the BR produced biomethane with a CH₄ content of 91.15 ± 1.01% under a gas retention time (GRT) of 12.7 h, while the TBR-P operation resulted in a CH₄ content of 90.92 ± 2.15% under a GRT of 6 h. The TBR-S operated at a lower GRT (4 h), yielding an effluent gas richer in CH₄ (93.08 ± 0.39%). Lowering the GRT further deteriorated the efficiency but did not influence the metabolic pathway, since no trace of volatile fatty acids was detected. These findings are essential indicators of the process stability under mesophilic conditions. Full article

The first discovery of methanogens led to the formation of a new domain of life known as Archaea. The Archaea domain exhibits properties vastly different from previously known Bacteria and Eucarya domains. However, for a certain multi-step process, a syntrophic relationship between organisms from all domains is needed. This process is called methanogenesis and is defined as the biological production of methane. Different methanogenic pathways prevail depending on substrate availability and the employed order of methanogenic Archaea. Most methanogens reduce carbon dioxide to methane with hydrogen through a hydrogenotrophic pathway. For hydrogen activation, a group of enzymes called hydrogenases is required. Regardless of the methanogenic pathway, electrons are carried between microorganisms by hydrogen. Naturally occurring processes, such as methanogenesis, can be engineered for industrial use. With the growth and emergence of new industries, the amount of produced industrial waste is an ever-growing environmental problem. For successful wastewater remediation, a syntrophic correlation between various microorganisms is needed. The composition of microorganisms depends on wastewater type, organic loading rates, anaerobic reactor design, pH, and temperature. The last step of anaerobic wastewater treatment is production of biomethane by methanogenesis, which is thought to be a cost-effective means of energy production for this renewable biogas. Full article

Anaerobic fermentation is a potentially cost-effective approach to disposing of metal-contaminated biowaste collected during phytoremediation. However, the compound heavy metals contained in the biowaste may limit the efficiency of anaerobic fermentation. In this study, anaerobic fermentation with alfalfa harvested from an iron tailing as the feedstock was set up and further enhanced by granular activated carbon (AC). The results showed that adding AC improved the cumulative biogas yields of alfalfa contaminated with metals (AM) by 2.26 times. At the biogas peak stage, plenty of microbes were observed on the surface of the AC, and the functional groups of AC contributed to better electron transfer, lower heavy metal toxicity and higher CH₄ contents. AC increased the richness and decreased the diversity of bacteria while reducing both the richness and diversity of archaea. The AC addition resulted in higher relative abundance of Prevotella_7, Bacteroides and Ruminiclostridium_1, which enhanced the hydrolysis of substrate and produced more precursors for methanogenesis. Meanwhile, the relative abundances of Methanosarcina and Methanobacterium were remarkably increased together with the metabolism of cofactors and vitamins, indicating the enhancement of both the acetoclastic and hydrotrophic methanogenesis. The present study provided new insights into the microbial responses of the anaerobic fermentation in heavy-metal-contaminated plants and proved the possibility of enhancing the biogas production by AC. Full article