Viral and Bacterial Community Dynamics in Food Waste and Digestate from Full-Scale Biogas Plants.

Anaerobic digestion (AD) is commonly used in food waste treatment. Prokaryotic microbial communities in AD of food waste have been comprehensively studied. The role of viruses, known to affect microbial dynamics and metabolism, remains largely unexplored. This study employed metagenomic analysis and recovered 967 high-quality viral bins within food waste and digestate derived from 8 full-scale biogas plants. The diversity of viral communities was higher in digestate. In silico predictions linked 20.8% of viruses to microbial host populations, highlighting possible virus predators of key functional microbes. Lineage-specific virus-host ratio varied, indicating that viral infection dynamics might differentially affect microbial responses to the varying process parameters. Evidence for virus-mediated gene transfer was identified, emphasizing the potential role of viruses in controlling the microbiome. AD altered the specific process parameters, potentially promoting a shift in viral lifestyle from lysogenic to lytic. Viruses encoding auxiliary metabolic genes (AMGs) were involved in microbial carbon and nutrient cycling, and most AMGs were transcriptionally expressed in digestate, meaning that viruses with active functional states were likely actively involved in AD. These findings provided a comprehensive profile of viral and bacterial communities and expanded knowledge of the interactions between viruses and hosts in food waste and digestate.

Treatment of azo dye-containing wastewater in a combined UASB-EMBR system: Performance evaluation and membrane fouling study.

This work investigated the treatment of azo dye-containing wastewater in an upflow anaerobic sludge blanket (UASB) reactor combined with an electro-membrane bioreactor (EMBR). Current densities of 20 A m-2 and electric current exposure mode of 6'ON/30'OFF were applied to compare the performance of the EMBR to a conventional membrane bioreactor (MBR). The results showed that dye (Drimaren Red CL-7B) removal occurred predominantly in the UASB reactor, which accounted for 57% of the total dye removal achieved by the combined system. When the MBR was assisted by electrocoagulation, the overall azo dye removal efficiency increased from 60.5 to 67.1%. Electrocoagulation batch tests revealed that higher decolorization rates could be obtained with a current density of 50 A m-2. Over the entire experimental period, the combined UASB-EMBR system exhibited excellent performance in terms of chemical oxygen demand (COD) and NH4+-N removal, with average efficiencies above 97%, while PO43--P was only consistently removed when the electrocoagulation was used. Likewise, a consistent reduction in the absorption spectrum of aromatic amines was observed when the MBR was electrochemically assisted. In addition to improving the pollutants removal, the use of electrocoagulation reduced the membrane fouling rate by 68% (0.25-0.08 kPa d-1), while requiring additional energy consumption and operational costs of 1.12 kWh m-3 and 0.32 USD m-3, respectively. Based on the results, it can be concluded that the combined UASB-EMBR system emerges as a promising technological approach for textile wastewater treatment.

Silver nanoparticles incorporated with superior silica nanoparticles-based rice straw to maximize biogas production from anaerobic digestion of landfill leachate.

Treating hazardous landfill leachate poses significant environmental challenges due to its complex nature. In this study, we propose a novel approach for enhancing the anaerobic digestion of landfill leachate using silver nanoparticles (Ag NPs) conjugated with eco-friendly green silica nanoparticles (Si NPs). The synthesized Si NPs and Ag@Si NPs were characterized using various analytical techniques, including transmission electron microscopy, X-ray diffraction, and Fourier-transform infrared spectroscopy. The anaerobic digestion performance of Si NPs and Ag@Si NPs was tested by treating landfill leachate samples with 50 mg/L of each NP. The results demonstrated an enhancement in the biogas production rate compared to the control phase without the nanocomposite, as the biogas production increased by 14% and 37% using Si NPs and Ag@Si NPs. Ag@Si NPs effectively promoted the degradation of organic pollutants in the leachate, regarding chemical oxygen demand (COD) and volatile solids (VS) by 58% and 65%. Furthermore, microbial analysis revealed that Ag@Si NPs enhanced the activity of microbial species responsible for the methanogenic process. Overall, incorporating AgNPs conjugated with eco-friendly green Si NPs represents a sustainable and efficient approach for enhancing the anaerobic digestion of landfill leachate.

Ultrasonication as anaerobic digestion pretreatment to improve sewage sludge methane production: Performance and microbial characterization.

A large amount of sludge is inevitably produced during sewage treatment. Ultrasonication (US) as anaerobic digestion (AD) pretreatment was implemented on different sludges and its effects on batch and semi-continuous AD performance were investigated. US was effective in sludge SCOD increase, size decrease, and CH4 production in the subsequent AD, and these effects were enhanced with an elevated specific energy input. As indicated by semi-continuous AD experiments, the mean daily CH4 production of US-pretreated A2O-, A2O-MBR-, and AO-AO-sludge were 176.9, 119.8, and 141.7 NmL/g-VSadded, which were 35.1%, 32.1% and 78.2% higher than methane production of their respective raw sludge. The US of A2O-sludge achieved preferable US effects and CH4 production due to its high organic content and weak sludge structure stability. In response to US-pretreated sludge, a more diverse microbial community was observed in AD. The US-AD system showed negative net energy; however, it exhibited other positive effects, e.g., lower required sludge retention time and less residual total solids for disposal. US is a feasible option prior to AD to improve anaerobic bioconversion and CH4 yield although further studies are necessary to advance it in practice.

An in-situ biochar-enhanced anaerobic membrane bioreactor for swine wastewater treatment under various organic loading rates.

A biochar-assisted anaerobic membrane bioreactor (BC-AnMBR) was conducted to evaluate the performance in treating swine wastewater with different organic loading rates (OLR) ranging from 0.38 to 1.13 kg-COD/(m3.d). Results indicated that adding spent coffee grounds biochar (SCG-BC) improved the organic removal efficiency compared to the conventional AnMBR, with an overall COD removal rate of > 95.01%. Meanwhile, methane production of up to 0.22 LCH4/gCOD with an improvement of 45.45% was achieved under a high OLR of 1.13 kg-COD/(m3.d). Furthermore, the transmembrane pressure (TMP) in the BC-AnMBR system was stable at 4.5 kPa, and no irreversible membrane fouling occurred within 125 days. Microbial community analysis revealed that the addition of SCG-BC increased the relative abundance of autotrophic methanogenic archaea, particularly Methanosarcina (from 0.11% to 11.16%) and Methanothrix (from 16.34% to 24.05%). More importantly, Desulfobacterota and Firmicutes phylum with direct interspecific electron transfer (DIET) capabilities were also enriched with autotrophic methanogens. Analysis of the electron transfer pathway showed that the concentration of c-type cytochromes increased by 38.60% in the presence of SCG-BC, and thus facilitated the establishment of DIET and maintained high activity of the electron transfer system even at high OLR. In short, the BC-AnMBR system performs well under various OLR conditions and is stable in the recovery energy system for swine wastewater.

Greenhouse gas emission potential of tropical coastal sediments in densely urbanized area inferred from metabarcoding microbial community data.

Although benthic microbial community offers crucial insights into ecosystem services, they are underestimated for coastal sediment monitoring. Sepetiba Bay (SB) in Rio de Janeiro, Brazil, holds long-term metal pollution. Currently, SB pollution is majorly driven by domestic effluents discharge. Here, functional prediction analysis inferred from 16S rRNA gene metabarcoding data reveals the energy metabolism profiles of benthic microbial assemblages along the metal pollution gradient. Methanogenesis, denitrification, and N2 fixation emerge as dominant pathways in the eutrophic/polluted internal sector (Spearman; p < 0.05). These metabolisms act in the natural attenuation of sedimentary pollutants. The methane (CH4) emission (mcr genes) potential was found more abundant in the internal sector, while the external sector exhibited higher CH4 consumption (pmo + mmo genes) potential. Methanofastidiosales and Exiguobacterium, possibly involved in CH4 emission and associated with CH4 consumers respectively, are the main taxa detected in SB. Furthermore, SB exhibits higher nitrous oxide (N2O) emission potential since the norB/C gene proportions surpass nosZ up to 4 times. Blastopirellula was identified as the main responsible for N2O emissions. This study reveals fundamental contributions of the prokaryotic community to functions involved in greenhouse gas emissions, unveiling their possible use as sentinels for ecosystem monitoring.

Biochar alleviates inhibition effects of humic acid on anaerobic digestion: Insights to performances and mechanisms.

To recover methane from waste activated sludge through anaerobic digestion (AD) is one promising alternative to achieve carbon neutrality for wastewater treatment plants. However, humic acids (HAs) are one of the major compositions in waste activated sludge, and their accumulation performs inhibition effects on AD. This study investigated the potentials of biochar (BC) in alleviating inhibition effects of HAs on AD. Results showed that although the accumulated HAs reduced methane yield by 9.37% compared to control, the highest methane yield, 132.6 mL CH4/g VSS, was obtained after adding BC, which was 45.9% higher than that in HA group. Mechanism analysis showed that BC promoted the activities of hydrolase such as protease and α-glucosidase, which were 69.7% and 29.7% higher than those in HA group, respectively. The conversion of short-chain fatty acids was accelerated. In addition, the evolution of electroactive microorganisms like Clostridium_sensu_stricto_13 and Methanosaeta were consistent with the activitiy of electron transfer and the content of cytochrome c. Furthermore, parts of HAs rather than all of them were adsorbed by BC, and the remaining free HAs and BC formed synergistic effects on methanogenesis, then both CO2 reduction and acetoclastic methanogenesis pathways were improved. The findings may provide some solutions to alleviate inhibition effects of HAs on AD.

Pilot-scale evaluation of cascade anaerobic digestion of mixed municipal wastewater treatment sludges.

This work assessed the performance of a pilot-scale cascade anaerobic digestion (AD) system when treating mixed municipal wastewater treatment sludges. The cascade system was compared with a conventional continuous stirred tank reactor (CSTR) digester (control) in terms of process performance, stability, and digestate quality. The results showed that the cascade system achieved higher volatile solids removal (VSR) efficiencies (28-48%) than that of the reference (25-41%) when operated at the same solids residence time (SRT) in the range of 11-15 days. When the SRT of the cascade system was reduced to 8 days the VSR (32-36%) was only slightly less than that of the reference digester that was operated at a 15-day SRT (39-43%). Specific hydrolysis rates in the first stage of the cascade system were 66-152% higher than those of the reference. Additionally, the cascade system exhibited relatively stable effluent concentrations of volatile fatty acids (VFAs: 100-120 mg/l), while the corresponding concentrations in the control effluent demonstrated greater fluctuations (100-160 mg/l). The cascade system's effluent pH and VFA/alkalinity ratios were consistently maintained within the optimal range. During a dynamic test when the feed total solids concentration was doubled, total VFA concentrations (85-120 mg/l) in the cascade system were noticeably less than those (100-170 mg/l) of the control, while the pH and VFA/alkalinity levels remained in a stable range. The cascade system achieved higher total solids (TS) content in the dewatered digestate (19.4-26.8%) than the control (17.4-22.1%), and E. coli log reductions (2.0-4.1 log MPN/g TS) were considerably higher (p < 0.05) than those in the control (1.3-2.9 log MPN/g TS). Overall, operating multiple CSTRs in cascade mode at typical SRTs and mixed sludge ratios enhanced the performance, stability digesters, and digestate quality of AD. PRACTITIONER POINTS: Enhanced digestion of mixed sludge digestion with cascade system. Increased hydrolysis rates in the cascade system compared to a reference CSTR. More stable conditions for methanogen growth at both steady and dynamic states. Improved dewaterability and E. coli reduction of digestate from the cascade system.

National Greenhouse Gas Emission Reduction Potential from Adopting Anaerobic Digestion on Large-Scale Dairy Farms in the United States.

Waste-to-energy systems can provide a functional demonstration of the economic and environmental benefits of circularity, innovation, and reimagining existing systems. This study offers a robust quantification of the greenhouse gas (GHG) emission reduction potential of the adoption of anaerobic digestion (AD) technology on applicable large-scale dairy farms in the contiguous United States. GHG reduction estimates were developed through a robust life cycle modeling framework paired with sensitivity and uncertainty analyses. Twenty dairy configurations were modeled to capture important differences in housing and manure management practices, applicable AD technologies, regional climates, storage cleanout schedules, and methods of land application. Monte Carlo results for the 90% confidence interval illustrate the potential for AD adoption to reduce GHG emissions from the large-scale dairy industry by 2.45-3.52 MMT of CO2-eq per year considering biogas use only in renewable natural gas programs and as much as 4.53-6.46 MMT of CO2-eq per year with combined heat and power as an additional biogas use case. At the farm level, AD technology may reduce GHG emissions from manure management systems by 58.1-79.8% depending on the region. Discussion focuses on regional differences in GHG emissions from manure management strategies and the challenges and opportunities surrounding AD adoption.

Detection of anaerobic and aerobic bacteria from commercial tattoo and permanent makeup inks.

Tattooing and use of permanent makeup (PMU) have dramatically increased over the last decade, with a concomitant increase in ink-related infections. Studies have shown evidence that commercial tattoo and PMU inks are frequently contaminated with pathogenic microorganisms. Considering that tattoo inks are placed into the dermal layer of the skin where anaerobic bacteria can thrive and cause infections in low-oxygen environments, the prevalence of anaerobic and aerobic bacteria should be assessed in tattoo and PMU inks. In this study, we tested 75 tattoo and PMU inks using the analytical methods described in the FDA Bacteriological Analytical Manual Chapter 23 for the detection of both aerobic and anaerobic bacterial contamination, followed by 16S rRNA gene sequencing for microbial identification. Of 75 ink samples, we found 26 contaminated samples with 34 bacterial isolates taxonomically classified into 14 genera and 22 species. Among the 34 bacterial isolates, 19 were identified as possibly pathogenic bacterial strains. Two species, namely Cutibacterium acnes (four strains) and Staphylococcus epidermidis (two strains) were isolated under anaerobic conditions. Two possibly pathogenic bacterial strains, Staphylococcus saprophyticus and C. acnes, were isolated together from the same ink samples (n = 2), indicating that tattoo and PMU inks can contain both aerobic (S. saprophyticus) and anaerobic bacteria (C. acnes). No significant association was found between sterility claims on the ink label and the absence of bacterial contamination. The results indicate that tattoo and PMU inks can also contain anaerobic bacteria. IMPORTANCE: The rising popularity of tattooing and permanent makeup (PMU) has led to increased reports of ink-related infections. This study is the first to investigate the presence of both aerobic and anaerobic bacteria in commercial tattoo and PMU inks under aerobic and anaerobic conditions. Our findings reveal that unopened and sealed tattoo inks can harbor anaerobic bacteria, known to thrive in low-oxygen environments, such as the dermal layer of the skin, alongside aerobic bacteria. This suggests that contaminated tattoo inks could be a source of infection from both types of bacteria. The results emphasize the importance of monitoring these products for both aerobic and anaerobic bacteria, including possibly pathogenic microorganisms.

New insight into enhanced carbon recovery from anaerobic fermentation of waste activated sludge with cation exchange resin coupled with NaCl pretreatment.

Carbon recovery from waste activated sludge has been attracting considerable attention. However, the migration and transformation patterns of carbon sources between the phases have rarely been reported. In this study, a novel strategy using cation exchange resin (CER) coupled with sodium chloride (NaCl) to enhance carbon recovery through anaerobic fermentation (AF) was proposed. The results demonstrated that CER coupled with NaCl destroyed OH and CO stretching in amide I while promoting the formation of ß-sheet and random coil structures, leading to sludge disintegration. This significantly improved the kinetics of endogenous carbon release, resulting in the release of 1146.33 mg/L of carbon from the solid sludge into the liquid phase. Approximately 75.61 % of the initial carbon source was bio-transformed into short-chain fatty acids. Correspondingly, carbon recovery was significantly increased up to 852.23 mg C/L, 4.57 times that of the control. Mechanism exploration revealed that carbon source recovery was significantly elevated by the synergistic effect of CER and NaCl. CER effectively removed high-valence cations from extracellular polymeric substance (EPS), weakening its bridging and adsorption-electro neutralization capabilities, promoting protein deflocculation, and triggering EPS disruption to release extracellular carbon sources. NaCl disrupted the ionic strength and distribution inside and outside microbial cells, creating an osmotic pressure difference that resulted in cell plasmolysis and lysis, ultimately inducing the release of intracellular carbon sources. Economic and carbon emission reduction benefit analyses verified that the CER coupled with NaCl pretreatment is a cost-effective sludge treatment strategy. This study illustrates the carbon source migration and transformation pathways in the CER coupled with NaCl-assisted AF process, providing guidance for sustainable sludge management.

Comparison of the diversity of anaerobic-cultured gut bacterial communities on different culture media using 16S rDNA sequencing.

The gut microbiome is a dense and diverse community of different microorganisms that deeply influence human physiology and that have important interactions with pathogens. For the correct antibiotic treatment of infections, with its twin goals of effective inhibition of the pathogen and limitation of collateral damage to the microbiome, the identification of infectious organisms is key. Microbiological culturing is still the mainstay of pathogen identification, and anaerobic species are among the most demanding bacterial communities to culture. This study aimed to evaluate the impact of growth media on the culture of an-aerobic bacteria from human stool samples. Stool samples from eight human subjects were cultured each on a yeast extract cysteine blood agar (HCB) and modified peptone-yeast extract-glucose (MPYG) plate and subjected to Illumina NGS analysis after DNA extraction and amplification. The results showed tight clustering of sequencing samples belonging to the same human subject. Various differences in bacterial richness and evenness could be observed between the two media, with HCB plates supporting the growth of a more diverse microbial community, and MPYG plates improving the growth rates of certain taxa. No statistical significance was observed between the groups. This study highlights the importance of choosing the appropriate growth media for anaerobic bacterial culture and adjusting culture conditions to target specific pathological conditions. HCB plates are suitable for standard microbiological diagnostics, while MPYG plates may be more appropriate for targeting specific conditions. This work emphasizes the role of next-generation sequencing in supporting future research in clinical microbiology.

Recent advancements in wastewater treatment via anaerobic fermentation process: A systematic review.

This manuscript delves into the realm of wastewater treatment, with a particular emphasis on anaerobic fermentation processes, especially dark, photo, and dark-photo fermentation processes, which have not been covered and overviewed previously in the literature regarding the treatment of wastewater. Moreover, the study conducts a bibliometric analysis for the first time to elucidate the research landscape of anaerobic fermentation utilization in wastewater purification. Furthermore, microorganisms, ranging from microalgae to bacteria and fungi, emphasizing the integration of these agents for enhanced efficiency, are all discussed and compared. Various bioreactors, such as dark and photo fermentation bioreactors, including tubular photo bioreactors, are scrutinized for their design and operational intricacies. The results illustrated that using clostridium pasteurianum CH4 and Rhodopseudomonas palustris WP3-5 in a combined dark-photo fermentation process can treat wastewater to a pH of nearly 7 with over 90% COD removal. Also, integrating Chlorella sp and Activated sludge can potentially treat synthetic wastewater to COD, P, and N percentage removal rates of 99%,86%, and 79%, respectively. Finally, the paper extends to discuss the limitations and future prospects of dark-photo fermentation processes, offering insights into the road ahead for researchers and scientists.

The effect of raw material (cow and chicken manure) and reactor type for improving and maximizing biogas production.

Biomass energy is a type of renewable energy and animal waste is one of the main resources for its production. The purpose of this study is to investigate the effect of raw material type (cow and chicken manure) and the type of reactor (digester) on the biogas produced by measuring the amount of methane in the product. Three types of digester (metal, simple PVC, and PVC with leachate rotation) with the same volume (10 L) were prepared. Equipment was installed on the digesters to measure the pH and volume of produced gas. The experiments were carried out in controlled temperature conditions (28-30 °C) and in two stages. The first experiment was to load the digesters with cow excrement, and the second experiment was to load the digesters with chicken excrement. In both experiments, the digesters were fed with 1.5 kg of animal manure and water with a ratio of 1:1. During a period of 60 days, the volume of biogas and methane produced was measured and recorded. The results showed that the amount of biogas produced from chicken waste is more than the amount obtained from cow waste. However, the amount of methane produced using cow excrement was more than that of chicken excrement. Also, the performance of PVC digester with leachate rotation was better than the other two digesters, which could be due to the mixing of raw materials in this type of digester.

Enhanced methane production from waste activated sludge by potassium ferrate combined with thermal hydrolysis pretreatment.

Anaerobic digestion of waste activated sludge (WAS) was one of the directions of sludge treatment, but how to effectively improve the production of methane as a resource product of anaerobic digestion of sludge still needs further research. The study examined how the combination of potassium ferrate (PF) and thermal hydrolysis (TH) pretreatment affected methane production from sludge. The results demonstrated a positive synergistic effect on methane production with PF-TH pretreatment. Specifically, by employing a 0.05 g/g TSS (total suspended solids) PF in conjunction with TH at 80 °C for 30 min, the methane yield increased from 170.66 ± 0.92 to 232.73 ± 2.21 mL/g VSS (volatile suspended solids). The co-pretreatment of PF and TH has been substantiated by mechanism studies to effectively enhance the disintegration and biodegradability of sludge. Additionally, the variation of microbial community revealed an enrichment of active microorganisms associated with anaerobic digestion after treated with PF + TH, resulting in a total abundance increase from 11.87 to 20.45% in the PF + TH group.

A training goal-oriented categorization model of high-intensity interval training.

There are various categorization models of high-intensity interval training (HIIT) in the literature that need to be more consistent in definition, terminology, and concept completeness. In this review, we present a training goal-oriented categorization model of HIIT, aiming to find the best possible consensus among the various defined types of HIIT. This categorization concludes with six different types of HIIT derived from the literature, based on the interaction of interval duration, interval intensity and interval:recovery ratio. We discuss the science behind the defined types of HIIT and shed light on the possible effects of the various types of HIIT on aerobic, anaerobic, and neuromuscular systems and possible transfer effects into competition performance. We highlight various research gaps, discrepancies in findings and not yet proved know-how based on a lack of randomized controlled training studies, especially in well-trained to elite athlete cohorts. Our HIIT "toolbox" approach is designed to guide goal-oriented training. It is intended to lay the groundwork for future systematic reviews and serves as foundation for meta-analyses.

The effects of aerobic gymnastics training on performance-related variables in an elite athlete: a 2-year follow-up study.

This study investigates individual performance adaptations on 2 years of training between European Aerobics Championships. An elite, 22-year-old aerobic gymnast performed postural coordination test, Y-Balance test, squat and countermovement jumps, 60 s test of repeated jumps, an isokinetic leg muscle strength test, and the Wingate test. Postural stability and flexibility improved in terms of increased distance achieved in the Y-Balance test in the anterior (by 6.3%), posteromedial (by 2%), and posterolateral (by 4.8%) directions. Lower limb muscular endurance also increased, which can be corroborated by a reduced fatigue index in the 60 s test of repeated jumps (from 42% to 27% after the 1st and to 22% after the 2nd year of training). In addition, mean power increased during dominant (by 23.2% at 60°/s and by 18.5% at 180°/s) and non-dominant leg extension (by 4.9% at 180°/s and by 15.5% at 300°/s), plus dominant leg flexion (by 2.0% at 60°/s and by 6.9% at 300°/s). Similarly, peak torque/body weight ratio increased during dominant (by 24.9% at 60°/s, by 11.5% at 180°/s, and by 2.1% at 300°/s) and non-dominant leg extension (by 0.5% at 60°/s and by 6.4% at 300°/s), plus dominant leg flexion (by 1.7% at 60°/s and by 5.4% at 300°/s). However, 2 years of training failed to show any significant improvements in the explosive power of lower limbs and anaerobic performance. These findings indicate that general aerobic gymnastics training without any specific inputs leads to performance adaptation, namely, in abilities closely related to competition routine (dynamic balance and strength endurance of lower limbs).

A pH-dependent shift of redox cofactor specificity in a benzyl alcohol dehydrogenase of aromatoleum aromaticum EbN1.

We characterise a reversible bacterial zinc-containing benzyl alcohol dehydrogenase (BaDH) accepting either NAD+ or NADP+ as a redox cofactor. Remarkably, its redox cofactor specificity is pH-dependent with the phosphorylated cofactors favored at lower and the dephospho-forms at higher pH. BaDH also shows different steady-state kinetic behavior with the two cofactor forms. From a structural model, the pH-dependent shift may affect the charge of a histidine in the 2'-phosphate-binding pocket of the redox cofactor binding site. The enzyme is phylogenetically affiliated to a new subbranch of the Zn-containing alcohol dehydrogenases, which share this conserved residue. BaDH appears to have some specificity for its substrate, but also turns over many substituted benzyl alcohol and benzaldehyde variants, as well as compounds containing a conjugated C=C double bond with the aldehyde carbonyl group. However, compounds with an sp3-hybridised C next to the alcohol/aldehyde group are not or only weakly turned over. The enzyme appears to contain a Zn in its catalytic site and a mixture of Zn and Fe in its structural metal-binding site. Moreover, we demonstrate the use of BaDH in an enzyme cascade reaction with an acid-reducing tungsten enzyme to reduce benzoate to benzyl alcohol. KEY POINTS: •Zn-containing BaDH has activity with either NAD + or NADP+ at different pH optima. •BaDH converts a broad range of substrates. •BaDH is used in a cascade reaction for the reduction of benzoate to benzyl alcohol.

Metatranscriptomics-guided genome-scale metabolic reconstruction reveals the carbon flux and trophic interaction in methanogenic communities.

BACKGROUND: Despite rapid advances in genomic-resolved metagenomics and remarkable explosion of metagenome-assembled genomes (MAGs), the function of uncultivated anaerobic lineages and their interactions in carbon mineralization remain largely uncertain, which has profound implications in biotechnology and biogeochemistry. RESULTS: In this study, we combined long-read sequencing and metatranscriptomics-guided metabolic reconstruction to provide a genome-wide perspective of carbon mineralization flow from polymers to methane in an anaerobic bioreactor. Our results showed that incorporating long reads resulted in a substantial improvement in the quality of metagenomic assemblies, enabling the effective recovery of 132 high-quality genomes meeting stringent criteria of minimum information about a metagenome-assembled genome (MIMAG). In addition, hybrid assembly obtained 51% more prokaryotic genes in comparison to the short-read-only assembly. Metatranscriptomics-guided metabolic reconstruction unveiled the remarkable metabolic flexibility of several novel Bacteroidales-affiliated bacteria and populations from Mesotoga sp. in scavenging amino acids and sugars. In addition to recovering two circular genomes of previously known but fragmented syntrophic bacteria, two newly identified bacteria within Syntrophales were found to be highly engaged in fatty acid oxidation through syntrophic relationships with dominant methanogens Methanoregulaceae bin.74 and Methanothrix sp. bin.206. The activity of bin.206 preferring acetate as substrate exceeded that of bin.74 with increasing loading, reinforcing the substrate determinantal role. CONCLUSION: Overall, our study uncovered some key active anaerobic lineages and their metabolic functions in this complex anaerobic ecosystem, offering a framework for understanding carbon transformations in anaerobic digestion. These findings advance the understanding of metabolic activities and trophic interactions between anaerobic guilds, providing foundational insights into carbon flux within both engineered and natural ecosystems. Video Abstract.

Metagenomic analysis reveals the mechanisms of biochar supported nano zero-valent iron in two-phase anaerobic digestion of food waste: microbial community, CAZmey, functional genes and antibiotic resistance genes.

The mechanisms of biochar supported nano zero-valent iron (BC/nZVI) on two-phase anaerobic digestion of food waste were investigated. Results indicated that the performance of both acidogenic phase and methanogenic phase was effectively facilitated. BC/nZVI with the amount of 120 mg/L increased methane production by 32.21%. In addition, BC/nZVI facilitated direct interspecies electron transfer (DIET) between Geobacter and methanogens. Further analysis showed that BC/nZVI increased the abundance of most CAZymes in acidogenic phase. The study also found that BC/nZVI had positive effects on metabolic pathways and related functional genes. The abundances of acdA and ackA in acidogenic phase were increased by 151.75% and 36.26%, respectively, and the abundances of pilA and TorZ associated with DIET were also increased. Furthermore, BC/nZVI mainly removed IMP-12, CAU-1, cmeB, ErmR, MexW, ErmG, Bla2, vgaD, MuxA, and cpxA from this system, and reduced the antibiotic resistance genes for antibiotic inactivation resistance mechanisms.