Direct Interspecies Electron Transfer Research Articles

Transcriptomic evidence for an energetically advantageous relationship between Syntrophomonas wolfei and Methanothrix soehngenii.

Syntrophic interactions are key in anaerobic food chains, facilitating the conversion of complex organic matter into methane. A typical example involves acetogenic bacteria converting fatty acids (e.g., butyrate and propionate), a process thermodynamically reliant on H2 consumption by microorganisms such as methanogens. While most studies focus on H2-interspecies transfer between these groups, knowledge on acetate cross-feeding in anaerobic systems is lacking. This study investigated butyrate oxidation by co-cultures of Syntrophomonas wolfei and Methanospirillum hungatei, both with and without the addition of the acetate scavenger Methanothrix soehngenii. Growth and gene expression patterns of S. wolfei and M. hungatei were followed in the two conditions. Although butyrate consumption rates remained constant, genes in the butyrate degradation pathway of S. wolfei were less expressed in the presence of M. soehngenii, including genes involved in reverse electron transport. Higher expression of a type IV-pili operon in S. wolfei hints to the potential for direct interspecies electron transfer between S. wolfei and M. soehngenii and an energetically advantageous relationship between the two microorganisms. Overall, the presence of the acetate scavenger M. soehngenii positively influenced the energy metabolism of S. wolfei and highlighted the relevance of including acetate scavengers when investigating syntrophic fatty acid degradation.

Unnatural Direct Interspecies Electron Transfer Enabled by Living Cell-Cell Click Chemistry.

Direct interspecies electron transfer (DIET) is essential for maintaining the function and stability of anaerobic microbial consortia. However, only limited natural DIET modes have been identified and DIET engineering remains highly challenging. Here, an unnatural DIET between Shewanella oneidensis MR-1 (SO, electron donating partner) and Rhodopseudomonas palustris (RP, electron accepting partner) was artificially established by a facile living cell-cell click chemistry strategy. By introducing alkyne- or azide-modified monosaccharides onto the cell outer surface of the target species, precise covalent connections between different species in high proximity were realized via a fast click chemistry reaction. Remarkably, upon covalent connection, outer cell surface C-type cytochromes mediated DIET between SO and RP was achieved and identified, although this was never realized naturally. Moreover, this connection directly shifted the natural H2 mediated interspecies electron transfer (MIET) to DIET between SO and RP, which delivered superior interspecies electron exchange efficiency. Therefore, this work demonstrated a naturally unachievable DIET and an unprecedented MIET shift to DIET accomplished by cell-cell distance engineering, offering an efficient and versatile solution for DIET engineering, which would extend our understanding of DIET and open up new avenue for DIET exploration and applications.

Unnatural Direct Interspecies Electron Transfer Enabled by Living Cell‐Cell Click Chemistry

Direct interspecies electron transfer (DIET) is essential for maintaining the function and stability of anaerobic microbial consortia. However, only limited natural DIET modes have been identified and DIET engineering remains highly challenging. Here, an unnatural DIET between Shewanella oneidensis MR‐1 (SO, electron donating partner) and Rhodopseudomonas palustris (RP, electron accepting partner) was artificially established by a facile living cell‐cell click chemistry strategy. By introducing alkyne‐ or azide‐modified monosaccharides onto the cell outer surface of the target species, precise covalent connections between different species in high proximity were realized via a fast click chemistry reaction. Remarkably, upon covalent connection, outer cell surface C‐type cytochromes mediated DIET between SO and RP was achieved and identified, although this was never realized naturally. Moreover, this connection directly shifted the natural H2 mediated interspecies electron transfer (MIET) to DIET between SO and RP, which delivered superior interspecies electron exchange efficiency. Therefore, this work demonstrated a naturally unachievable DIET and an unprecedented MIET shift to DIET accomplished by cell‐cell distance engineering, offering an efficient and versatile solution for DIET engineering, which would extend our understanding of DIET and open up new avenue for DIET exploration and applications.

Improving anaerobic digestion performance after severe acidification: Unveiling the impacts of Fe3O4-bentonite composites in co-digestion of waste activated sludge and food waste.

Acidification recovery in anaerobic digestion of food waste is challenging. This study explored its in-situ recovery using a co-substrate of food waste and waste activated sludge. Fe3O4 and bentonite were used as conductor and carrier, respectively, to enhance AD performance under severe acidification. The application of Fe3O4-bentonite resulted in a 152% increase in cumulative methane in the Fe3O4-bentonite 10 digester, demonstrating its effectiveness in restoring the acidified AD system. In acidified systems, bentonite enhanced the diversity and richness of microbial communities due to its buffering capacity. The excessive non-conductive polysaccharides excreted by bacteria in extracellular polymeric substances reduced the possibility of electron transfer by Fe3O4. However, in the synergistic application of Fe3O4 and bentonite, this resistance was alleviated, increasing the possibility of direct interspecies electron transfer, and accelerating the consumption of volatile fatty acids. This approach of integrating carrier and conductive materials is significant for in-situ restoration of acidified systems.

Plasticizers inhibit food waste anaerobic digestion performance by affecting microbial succession and metabolism

The widely existed plastic additive plasticizers in organic wastes possibly pose negative influence on anaerobic digestion (AD) performance, the direct evidence about the effects of plasticizers on AD performance is still lacking. This study evaluated the influencing mechanism of two typical plasticizers bisphenol A (BPA) and dioctyl phthalate on the whole AD process. Results indicated that plasticizers addition inhibited methane production, and the inhibiting effects were reinforced with the increase of concentration. By contrast, 50mg/L BPA exhibited the strongest inhibition on methane production. Physicochemical analysis showed plasticizers inhibited the metabolism efficiency of soluble polysaccharide and volatile fatty acids. Microbial communities analyses suggested that plasticizers inhibited the direct interspecies electron transfer participators of methanogenic archaea (especially Methanosarcina) and syntrophic bacteria. Furthermore, plasticizers inhibited the methane metabolisms, key coenzymes (CoB, CoM, CoF420 and methanofuran) biosynthesis and the metabolisms of major organic matters. This study shed light on the effects of plasticizers on AD performance and provided new insights for assessing the influences of plasticizers or plastic additives on the disposal of organic wastes.

Magnetite particles accelerate methanogenic degradation of highly concentrated acetic acid in anaerobic digestion process

The anaerobic digestion (AD) process has become significant for its capability to convert organic wastewater into biogas, a valuable energy source. Excessive acetic acid accumulation in the anaerobic digester can inhibit methanogens, ultimately leading to the deterioration of process performance. Herein, the effect of magnetite particles (MP) as an enhancer on the methanogenic degradation of highly-concentrated acetate (6 g COD/L) was examined through long-term sequential AD batch tests. Bioreactors with (AM) and without (AO) MP were compared. AO experienced inhibition and its methane production rate (qm) converged to 0.45 L CH4/g VSS/d after 10 sequential batches (AO10, the 10th batch in a series of the sequential batch tests conducted using bioreactors without MP addition). In contrast, AM achieved 3–425% higher qm through the sequential batches, indicating that MP could counteract the inhibition caused by the highly-concentrated acetate. MP addition to inhibited bioreactors (AO10) successfully restored them, achieving qm of 1.53 L CH4/g VSS/d, 3.4 times increase from AO10 after 8 days lag time, validating its potential as a recovery strategy for inhibited digesters with acetate accumulation. AM exhibited higher microbial populations (1.8–3.8 times) and intracellular activity (9.3 times) compared to AO. MP enriched Methanosaeta, Peptoclostridium, Paraclostridium, OPB41, and genes related to direct interspecies electron transfer and acetate oxidation, potentially driving the improvement of qm through MP-mediated methanogenesis. These findings demonstrated the potential of MP supplementation as an effective strategy to accelerate acetate-utilizing methanogenesis and restore an inhibited anaerobic digester with high acetate accumulation.

Research progress and perspective on sludge anaerobic digestion technology: A bibliometric analysis

Abstract Rational disposal of sludge is an ongoing concern. This work is the first attempt for in-depth statistical analysis of anaerobic digestion (AD) research in recent three decades (1986–2022) using both quantitative and qualitative approaches in bibliometrics to investigate the research progress, trends and hot spots. All publications in the Web of Science Core Collection database from 1986 to April 4, 2022 were analyzed. Results showed that the research on AD started in 1999 and the number of papers significantly increased since 2012. The research about the disposal of sewage sludge mainly focuses on energy recovery (e.g. methane and short chain volatile organic acids) by AD. Besides, different pretreatment technologies were studied in this study to eliminate the negative effects on the disposal of sludge caused by hydrolysis (rate-limiting step of AD), water content (increasing the costs) and heavy metal (toxic to the environment) of sludge. Of those, the treatment technologies related to direct interspecies electron transfer were worth further studied in the future. Towards that end, iron conductive material, iron-based advanced oxidation and biological treatment were concluded as the prospective technologies and worth to further study.

Role of exogenous hydrogen in methane yield during anaerobic digestion of waste fat, oil, and grease by coupling carbon cloth with exogenous hydrogen

Carbon cloth (CC) coupled with exogenous hydrogen (EH2) was used to promote direct interspecies electron transfer (DIET) during anaerobic digestion of waste fat, oil, and grease (FOG). However, whether the change of EH2 injection pattern would further enhance DIET and FOG conversion for methane production is unknown, and the underlying mechanism is not fully understood. In this study, three EH2 injection pressures (0.2/0.4/0.6 atm) and frequencies (once/every 8 days/every 4 days) were chosen to collaborate with CC for investigating the role of EH2 in methane yield during FOG digestion. The results show that, combined with CC, increased EH2 injection pressure/frequency resulted in gradually enhanced DIET to increase FOG conversion rate (from 69.9% to 96.7%), leading to the highest cumulative methane production (1204.9 mL/gVSadded) 58.9% higher than that of Control. During the digestion, the secretion of extracellular polymeric substances was increased by EH2 to promote bioaggregation and electron transfer between cells for enhanced biological DIET in the digestate. The influence of EH2 on the microbial communities on CC surface was bigger than that in suspended sludge, leading to enhanced syntrophic acetate oxidation-hydrogenotrophic methanogenesis (SAO-HM) for acetate conversion on CC. As the amount of EH2 increased, the acetate conversion by SAO-HM via DIET was improved, which simultaneously promoted the syntrophic metabolisms between acidogens/acetogens and methanogens to achieve more efficient FOG conversion. Together, this work provides new insight into the role of EH2 in the enhancement of DIET and methane yield during FOG digestion.

Microbial synergies drive simultaneous biodegradation of ethoxy and alkyl chains of Nonylphenol Ethoxylate in fluidized bed reactors

The widely-used surfactant Nonylphenol Ethoxylate (NPEO) produces endocrine-disrupting compounds during biodegradation, with these byproducts being more harmful than untreated NPEO. This study investigates the effectiveness of a Fluidized Bed Reactor (FBR) in reducing the production of 4-Nonylphenol (4-NP) during the biodegradation of NPEO. Two identical FBR filled with sand were used to assess the NPEO degradation and to enhance the microbial consortia capable of breaking down the complex byproducts, ethanol and fumarate were introduced as co-substrates. Our findings demonstrate the significant potential of the FBR, especially when coupled with fumarate, for enhancing the surfactant degradation. It outperforms the efficiency achieved with ethanol as the primary electron donor, albeit with a higher rate of byproduct production. Microbial community taxonomy and metabolic prediction revealed the high abundance of Geobacter (1.51–31.71%) and Methanobacterium (1.08–13.81%) in non-conductive sand. This may hint a new metabolic interaction and expand our understanding of Direct Interspecies Electron Transfer (DIET) in bioreactors applied to micropollutants degradation. Such an intricate relationship between facultative and anaerobes working together to simultaneously biodegrade the ethoxy and alkyl chains presents a new perspective on NPEO degradation and can potentially be extended to other micropollutants.

Understanding of the potential role of carbon dots in promoting interspecies electron transfer process in anaerobic co-digestion under magnetic field: Focusing on methane and hydrogen production

Anaerobic digestion shows great potential for biomass energy recovery, however, the low electron transfer efficiency between microorganisms limits overall anaerobic digestion performance. In this work, aloe peel-derived carbon dots (carbon quantum dots, CQDs) were designed and investigated as accelerants for anaerobic digestion systems under an applied magnetic field to promote the interspecific electron transfer (IET) efficiency in methane and hydrogen production processes. Compared with the control group (188.9 mL/g VS, 4.2 mL/g VS, 34.7 %, and 26.3 %), the applied magnetic field (5 mT) significantly increased methane yield (256.1 mL/g VS), hydrogen yield (7.0 mL/g VS), total chemical oxygen demand reduction (41.7 %), and energy conversion efficiency (35.7 %). The introduction of the CQDs (0.18–0.45 wt.%) under a magnetic field (5 mT) has further positive effects on digestion performance. The experimental group incorporating 0.36 wt.% CQDs under a 5 mT magnetic field obtained the highest methane yield (348.9 mL/g VS), hydrogen yield (9.9 mL/g VS), and total chemical oxygen demand reduction (58.4 %), and thus the energy recovery efficiency was increased by 85 % compared with control group. The digestate incorporating 0.36 wt.% CQDs exhibited excellent thermal stability (59.71 %) and total nutrient content (38.17 g/kg), and has great potential as a main ingredient in compound fertilizer. The potential role of the magnetic field and CQDs in enhancing hydrogen interspecies electron transfer (HIET) and direct interspecies electron transfer (DIET) for methane and hydrogen production is further illustrated, providing a feasible solution for improving energy recovery from biomass.

A new strategy for accelerating recovery of anaerobic granular sludge after low-temperature shock: In situ regulation of quorum sensing microorganisms embedded in polyvinyl alcohol sodium alginate

Low-temperature could inhibit the performance of anaerobic granular sludge (AnGS). Quorum sensing (QS), as a communication mode between microorganisms, can effectively regulate AnGS. In this study, a kind of embedded particles (PVA/SA@Serratia) based on signal molecule secreting bacteria was prepared by microbial immobilization technology based on polyvinyl alcohol and sodium alginate to accelerate the recovery of AnGS system after low temperature. Low-temperature shock experiment verified the positive effect of PVA/SA@Serratia on restoring the COD removal rate and methanogenesis capacity of AnGS. Further analysis by metagenomics analysis showed that PVA/SA@Serratia stimulated higher QS activity and promoted the secretion of extracellular polymeric substance (EPS) in AnGS. The rapid construction of EPS protective layer effectively accelerated the establishment of a robust microbial community structure. PVA/SA@Serratia also enhanced multiple methanogenic pathways, including direct interspecies electron transfer. In conclusion, this study demonstrated that PVA/SA@Serratia could effectively strengthen AnGS after low-temperature shock.

Coupling of biogas residue biochar and low-magnitude electric fields promotes anaerobic co-digestion of sewage sludge and food waste.

Biochar-assisted anaerobic digestion (AD) remains constrained due to the inefficient decomposition of complex organics, even with the direct interspecies electron transfer (DIET) pathway. The coupling of electrochemistry with the anaerobic biological treatment could shorten lengthy retention time in co-digestion by improving electron transfer rates and inducing functional microbial acclimation. Thus, this work investigated the potential of improving the performance of AD by coupling low-magnitude electric fields with biochar derived from the anaerobically digested biogas residue. Different voltages (0.3, 0.6, and 0.9 V) were applied at various stages to assess the impact on biochar-assisted AD. The results indicate that an external voltage of 0.3 V, coupled with 5 g/L of biochar, elevates CH4 yield by 45.5% compared to biogas residue biochar alone, and the coupled approach increased biogas production by up to 143% within 10 days. This finding may be partly explained by the enhanced utilization of substrates and the increased amounts of specific methanogens such as Methanobacterium and Methanosarcina. The abundance of the former increased from 4.0 to 11.3%, which enhances the DIET between microorganisms. Furthermore, the coupling method shows better potential for enhancing AD compared to preparing iron-based biochar, and these results present potential avenues for its broader applications.

Allophane improves anaerobic digestion of chicken manure by alleviating ammonia inhibition and intensifying direct interspecies electron transfer

Synthesized allophane was employed in anaerobic digestion of chicken manure to improve the stability and methane production under ammonia inhibition. Adding 0.5 %, 1.0 % and 1.5 % (w/w) allophane increased the methane production by 261 ∼ 350 % compared with the group without allophane addition. Further investigation indicated that the maximum adsorption capacity of allophane for NH4+-N achieved at 261.9 mg/g; it suggested that allophane adsorption potentially alleviated the ammonia inhibition, which also was reflected by the increase in the activity of the related enzyme, such as coenzyme F420. Moreover, allophane addition also intensified the direct interspecies electron transfer (DIET) in anaerobic digestion; it can be well supported by the increased relative abundance of Methanosaeta and Methanosarcina involved in the DIET. Overall, the improved anaerobic digestion via alleviating ammonia inhibition and intensifying DIET by allophane was elucidated comprehensively, which can contribute to the development of a functional additive for efficient anaerobic digestion in practical application

Different oxygen affinities of methanotrophs and Comammox Nitrospira inform an electrically induced symbiosis for nitrogen loss

Aerobic methanotrophs establish a symbiotic association with denitrifiers to facilitate the process of aerobic methane oxidation coupled with denitrification (AME-D). However, the symbiosis has been frequently observed in hypoxic conditions continuing to pose an enigma. The present study has firstly characterized an electrically induced symbiosis primarily governed by Methylosarcina and Hyphomicrobium for the AME-D process in a hypoxic niche caused by Comammox Nitrospira. The kinetic analysis revealed that Comammox Nitrospira exhibited a higher apparent oxygen affinity compared to Methylosarcina. While the coexistence of comammox and AME-D resulted in an increase in methane oxidation and nitrogen loss rates, from 0.82±0.10 to 1.72±0.09 mmol CH4 d−1 and from 0.59±0.04 to 1.30±0.15 mmol N2 d−1, respectively. Furthermore, the constructed microbial fuel cells demonstrated a pronounced dependence of the biocurrents on AME-D due to oxygen competition, suggesting the involvement of direct interspecies electron transfer in the AME-D process under hypoxic conditions. Metagenomic and metatranscriptomic analysis revealed that Methylosarcina efficiently oxidized methane to formaldehyde, subsequently generating abundant NAD(P)H for nitrate reduction by Hyphomicrobium through the dissimilatory RuMP pathway, leading to CO2 production. This study challenges the conventional understanding of survival mechanism employed by AME-D symbionts, thereby contributing to the characterization responsible for limiting methane emissions and promoting nitrogen removal in hypoxic regions.

Enhancing microbial viability with biochar for increased methane production during the anaerobic digestion of chicken manure

This investigation explores the effects of integrating acid-alkali treated biochar into the anaerobic digestion of chicken manure. Digesters containing chicken manure with biochar addition exhibited an 8.4-fold improvement in methane production compared to untreated digesters. In addition, the porous structure of the biochar promoted microbial acclimation, facilitated direct interspecies electron transfer, and provided anchoring points for immobilisation. Anaerobic digesters containing chicken manure amended with biochar also demonstrated increased chemical oxygen demand removal, up to 81.8 %, and maintained lower total ammonia concentrations (2,478 mg/L) compared to levels in control reactors (4418 mg/L). In terms of the microbial community, the addition of propidium monoazide minimised errors in live microbial population estimation, providing new insights into the capacity of biochar to shield microbes from ammonia stress. These findings highlight biochar's quantitative benefits, emphasising its potential in enhancing methane production, mitigating ammonia stress, and promoting microbial resilience in anaerobic digestion.

Elucidating the development of cooperative anode-biofilm-structures

Microbial electrochemical systems are a highly versatile platform technology with a particular focus on the interplay of chemical and electrical energy conversion and offer immense potential for a sustainable bioeconomy. The industrial realization of this potential requires a critical focus on biofilm optimization if performance is to be controlled over a long period of time. Moreover, the aspect and influence of cooperativity has to be addressed as many applied anodic bioelectrochemical systems will most likely be operated with a diversity of interacting microbial species. Hence, the aim of this study was to analyze how interspecies dependence and cooperativity of a model community influence the development of anodic biofilms. To investigate biofilm activity in a spatially resolved manner, a microfluidic bioelectrochemical flow cell was developed that can be equipped with user-defined electrode materials and operates under laminar flow conditions. With this infrastructure, the development of single and co-culture biofilms of the two model organisms Shewanella oneidensis and Geobacter sulfurreducens on graphite electrodes was monitored by optical coherence tomography analysis. The interdependence in the co-culture biofilm was achieved by feeding the community with lactate, which is converted by S. oneidensis into acetate, which in turn serves as substrate for G. sulfurreducens. The results show that co-cultivation resulted in the formation of denser biofilms than in single culture. Moreover, we hypothesize that S. oneidensis in return utilizes the conductive biofilm matrix build by G. sulfurreducens for direct interspecies electron transfer (DIET) to the anode. FISH analysis revealed that the biofilms consisted of approximately two-thirds G. sulfurreducens cells, which most likely formed a conductive 3D network throughout the biofilm matrix, in which evenly distributed tubular S. oneidensis colonies were embedded without direct contact to the anode surface. Live/dead staining shows that the outermost biofilm contained almost exclusively dead cells (98 %), layers near the anode contained 45–56 % and the entire biofilm contained 82 % live cells. Our results exemplify how the architecture of the exoelectrogenic biofilm dynamically adapts to the respective process conditions.

Iron-mediated DAMO–anammox process: Revealing the mechanism of electron transfer

The nitrate denitrifying anaerobic methane oxidation–anaerobic ammonia oxidation (DAMO–anammox) can accomplish nitrogen removal and methane (CH4) reduction. This process greatly contributes to carbon emission mitigation and carbon neutrality. In this study, we investigated the electron transfer process of functional microorganisms in the iron-mediated DAMO–anammox system. Fe3+ could be bound to several functional groups (-CH3, COO-, –CH) in extracellular polymeric substance (EPS), and the functional groups bound were different at different iron concentration. Fe3+ underwent reduction reactions to produce Fe2+. Most Fe3+ and Fe2+ react with microorganisms and formed chelates with EPS. Three-dimensional fluorescence spectra showed that Fe3+ affected the secretion of tyrosine and tryptophan, which were essential for cytochrome synthesis. The presence of Fe3+ accelerated c-type cytochrome-mediated extracellular electron transfer (EET), and when more Fe3+ existed, the more cytochrome C expressed. DAMO archaea (M. nitroreducens) in the system exhibited a high positive correlation with the functional genes (resa and ccda) for cytochrome c synthesis. Some denitrifying microorganisms showed positive correlations with the abundance of riboflavin. This finding showed that riboflavin secreted by functional microorganisms acted as an electron shuttle. In addition, DAMO archaea were positively correlated with the hair synthesis gene pily1, which indicated that direct interspecies electron transfer (DIET) may exist in the iron-mediated DAMO–anammox system.

Material and microbial perspectives on understanding the role of biochar in mitigating ammonia inhibition during anaerobic digestion

With the increasing adoption of carbon-based strategies to enhance methanogenic processes, there is a growing concern regarding the correlation between biochar properties and its stimulating effects on anaerobic digestion (AD) under ammonia inhibition. This study delves into the relevant characteristics and potential mechanisms of biochar in the context of AD system under ammonia inhibition. The introduction of optimized biochar, distinguished by rich CO bond, abundant defect density, and high electronic capacity, resulted in a significant reduction in the lag period of anaerobic digestion system under 5.0 g/L ammonia stress, approximately by around 63 % compared to the control one. Biochar helps regulate the community structure, promotes the accumulation of acetate-consuming bacteria, in the AD system under ammonia inhibition. More examinations show that biochar promotes direct interspecies electron transfer in AD system under ammonia inhibition, as evidenced by diminished levels of bound electroactive extracellular polymeric substances, increased abundance of electroactive bacteria, and notably, the up-regulation of direct interspecies electron transfer associated genes, including the conductive pili and Cytochrome C genes, as revealed by meta-transcriptomic analysis. Additionally, gene expression related to proteins associated with ammonium detoxification were found to be up-regulated in systems supplemented with biochar. These findings provide essential evidence and insights for the selection and potential engineering of effective biochar to enhance AD performance under ammonia inhibition.

Exploring nature's filters: Peat-mineral mix for low and high-strength oilfield produced water reclamation

Nature-based solutions are encouraged for treating oilfield produced water from oil and gas extraction, a crucial undertaking that aligns with the Canadian oil sands industry's ambitious goal of zero waste, and the globally recognized Sustainable Development Goals (SDGs) pertaining to water conservation and ecosystem preservation. This study explored the use of peat-mineral mix (PMM), a leftover of inevitable oil sands mining, for treating low and high-strength wastewaters during biofiltration, which contained large molecular weight (44.3 kDa), which include alcohols, aliphatics, aromatics, and ketones, and can impart high toxicity to both fauna and flora (MicroTox: 99 %). The breakthrough curve indicated an effective initial adsorption phase driven by advection within the column dynamics. For complete organics removal and mechanistic insights, the wastewater was re-circulated in a continuous mode for up to 42 days. Here, we found that chemical oxygen demand was reduced from ∼85,000 mg/L to ∼965 mg/L). Kinetics investigations along with physicochemical characterization of PMM and wastewater suggested that chemisorption and anaerobic digestion contributed to the overall removal of contaminants. Chemisorption, led by hydrogen bonding and hydrophobic interactions, was the dominant mechanism, with a limited contribution from physical adsorption (surface area: 2.85 m2/g). The microbial community within the PMM bed was rich/diverse (Shannon > 6.0; Chao1 > 600), with ∼ 50 % unclassified phylotypes representing ‘microbial dark matter’. High electric conductivity (332.1 μS cm−1) of PMM and the presence of Geobacter, syntrophs, and Methanosaeta suggest that direct interspecies electron transfer was likely occurring during anaerobic digestion. Both low and high-strength wastewaters showed effective removal of dissolved organics (e.g., naphthenic acids, acid extractable fraction, oil and grease content), nutrients, and potentially toxic metals. The successful use of PMM in treating oilfield produced water offers promising avenues for embracing nature-based remediation solutions at oil refining sites.

Biochar-mediated synergistic promotion of DIET and IHT: Kinetic and thermodynamic insights into propionate two-step syntrophic methanogenesis

Biochar can effectively enhance the thermophilic anaerobic degradation of propionate. Nevertheless, previous studies frequently ascribed this enhancement to direct interspecies electron transfer (DIET) without considering the underlying kinetics and thermodynamics involved. Additionally, quantitative evaluation of biochar’s impact on interspecies electron transfer (IET) has been limited. In this study, we developed and optimized models for analyzing thermodynamics and electron transfer flux to elucidate the mechanism by which biochar enhances the two-step syntrophic methanogenesis of propionate. Findings revealed that biochar primarily facilitated DIET (1.78 × 10−4 A), leveraging its redox capacity. Additionally, it synergistically enhanced interspecies hydrogen transfer (4.08 × 10−10 A), significantly improving IET efficiency. This synergistic effect led to accelerated oxidative degradation rates of propionate (39.2 %) and intermediate acetate (13.5 %), expanding the thermodynamic windows for their syntrophic methanogenesis (221 % and 103 %, respectively). Accordingly, biochar increased the maximum rate of propionate methanation (71.9 %) and reduced the lag phase time (51.1 %). Moreover, biochar promoted the specific growth rates of acetogenic bacteria and methanogens while significantly increasing the abundance of Pelotomaculum, Methanoculleus, and Methanosarcina. These findings provide new insights and theoretical support for the use of biochar in promoting anaerobic digestion and facilitating IET during the process.