Pathway For Methane Production Research Articles

Lipase pretreatment enhances biochar‐assisted anaerobic digestion of food waste

AbstractAnaerobic digestion is a promising technology for the treatment of food waste (FW) but it requires optimization to improve its performance. This study investigated whether combining the addition of biochar with lipase pretreatment could improve the anaerobic digestion of FW. The results showed that biochar stimulated the release of soluble substances, resulting in an increase in the methane yield of groups with added biochar by 16.24% to 19.36% in comparison with the group without biochar. Lipase pretreatment substantially shortened the fermentation time required to complete the anaerobic digestion, from 15 to 9 days. Lipase pretreatment further improved microbial abundance and promoted the enrichment of functional microbes in the anaerobic digestion of FW mediated by biochar. It also changed the methane production pathway from acetotrophic to hydrogenotrophic, thereby ensuring the stability of the anaerobic digestion system in the presence of a high ammonia nitrogen concentration.

A meta-analytical approach for estimating methane suppression from dietary additives in ruminants

IntroductionMethane production inhibitors included in feed additives are increasingly used to suppress methane production in the rumen. Most products interact with the rumen microbiome and/or component biochemical pathways that scavenge free hydrogen ions (H+) to make methane (CH4 ). The capacity of these agents to inhibit rumen methane production is determined by the ability of the chemical to block methane production pathways, the amount of agent delivered to the rumen (i.e. the dose) and the absorption, distribution, metabolism and excretion (i.e. the pharmacokinetic) characteristics of the chemical that contribute to removal from the site of action (the rumen). The intrinsic inhibitory capacity of an agent determines the maximum rate of methane suppression. This maximal rate may reduce according to pharmacokinetic effects arising from dose rate and frequency. Most studies of additive methane reduction efficacy use total mixed ration (TMR; with the additive included into the ration) feeding systems and estimate methane reduction (absolute or relative) across a 24-hour period. Few studies report critical pharmacokinetic parameters, making it difficult to extrapolate findings into non-TMR systems (such as grazing) where differing doses and dose frequencies apply.MethodsWe consider the likely behaviour of a rumen-acting oral additive to reduce methane production applying basic pharmacokinetic principles to propose an analytical approach to data from multiple field studies employing different dose rates and dose frequencies to estimate methane suppression responses. This is based upon a logistic transformation of relative efficacy (percentage reduction in methane comparing treatment with control groups) as the dependent variable and includes total dose, dose frequency, quadratic and interaction terms between total dose and dose frequency as independent variables that potentially capture any pharmacokinetic effects on performance. The model was tested using simulation and verified against real data (cattle 3-nitrooxypropanol (3-NOP) methane-reduction studies).ResultsGood fit between predicted and observed methane suppression was obtained.DiscussionWe propose use of the general form of this logistic model with dose and dose frequency components included that address basic pharmacological impacts on methane suppression as the standard approach to meta-analysis.

Illuminating bioprocess responses to metal-based nanoparticles addition along hydrogen and methane production pathways: A review.

Recent research has discussed the positive impacts of metal-based nanoparticles (NPs) on bioprocesses producing either hydrogen (H2) or methane (CH4). The enhancement has been explained by mechanisms such as direct interspecies electron transfer (DIET), metal corrosion, and dissimilatory reduction. Such interactions could induce further benefits, such as controlling oxidation-reduction potential (ORP), mitigating toxicants, promoting enzymatic activity, and altering the microbiome, which have not yet been comprehensively discussed. Factors like metal type, oxidation state, and size of NPs are crucial for their reactivity and corresponding responses. This review discusses how different redox potentials of metals can regulate metabolic pathways and how NPs and their reactive ions can eliminate toxicants (e.g., sulfate) and enhance the activity of intra- and extracellular enzymes. The enrichment of responsive microorganisms in correlation with NPs is further discussed. A better understanding of the multifaceted role of metal-based NPs can guide potential new incorporation strategies to improve bioprocesses.

Microbial transformation of lignite into methane: Insights from anaerobic-activated sludge systems

Lignite pretreated by H2O2 was degraded by microorganisms in anaerobic activated sludge to produce methane, and the microorganisms were analyzed by high-throughput techniques. Samples fermented for 60 days were analyzed by high performance liquid chromatography (HPLC) and gas chromatography–mass spectrometry (GC–MS). Methane production was found to reach a maximum rate of 2.25 mL/g·d−1 at pH 7 and 35 °C, which was synchronized with the efficient consumption of organic acids, with a peak organic acid production of 202.13 COD/mg·L−1. A total of 37 compounds including alkanes, ketones, esters and polycyclic aromatic hydrocarbons were detected in the fermentation broth, and the species compositions of anaerobic sludge microbiota were fungal, bacterial and archaeal are Ascomycota, Bacteroidetes and Methanosaeta respectively in the fermentation system. Based on the analysis of above results, it is initially speculated that the biochemical pathway for lignite degradation and methane production by anaerobic sludge microorganisms is the acetate pathway.

The archaeal community in sediments of freshwater lakes of north-east Antarctic Peninsula: Structure and diversity

This research represents the first attempt to study the structure and diversity of the archaeal and methanogenic archaeal community in selected lakes around the Czech polar station J.G. Mendel on James Ross Island (JRI), Antarctica. Sediment samples from a total of 19 of JRI and the nearby Vega Island and Long Island, were analyzed using 16S rRNA and mcrA genes sequencing and real-time qPCR. Contrary to the sequences retrieved by 16S rRNA analysis, many more reads belonging to methanogens were found with mcrA gene sequencing. Generally, archaea represented only a small proportion (0–8.8%) of the total prokaryotic community. With the exception of lakes in the Solorina Valley and Lagoons Mesa area and Lake Esmeralda on Vega Island, methanogenic archaea made up a small proportion of the archaea present in most lakes. The genera Methanothrix and Methanosarcina were identified as the predominant methanogenic representatives in the lake samples. Sequences of representatives belonging to Methanothermobacter sp. and Methanomassiliicoccales and a high proportion of sequences belonging to Methanoperedens-like archaea, methanotrophs that associate anaerobic methane oxidation with denitrification, were recorded for the first time in Antarctica. It is also the first time that the presence of the genus Methanobacterium has been detected to such a large extent. Generally, sequences of the methanogens which might be involved in all three pathways of methane production were found in our samples, indicating the broad metabolic potential of the methanogens present. Individual lakes from one area shared much higher similarity in their methanogenic diversity with the lakes from another area rather than with the lakes within the same area, suggesting that a lake location is probably not the main factor influencing the diversity of the methanogens. Indeed, archaeal and methanogenic community structure and mcrA gene copy numbers varied even within a single lake, suggesting that more sampling within a single lake, preferably at different times of the year, will be necessary in the future for more comprehensive information. Although this is an initial study, our research unambiguously provides evidence that the lakes of the JRI and surrounding islands may be potential sources of new archaeal species or metabolic pathways.

Enhanced anaerobic digestion of food waste by the addition of cobalt iron oxide

In this study, the impact of cobalt iron oxide(CoFe2O4) on the anaerobic digestion (AD) of food waste (FW) was investigated including the variations in pH, VFAs, ORP, Fe2+/Fe3+ concentrations, SCOD, dehydrogenase activity, methane production and microbial community structure. The results showed that CoFe2O4 had a positive effect on the anaerobic digestion of food waste. CoFe2O4 could promote the release of SCOD, increase the dehydrogenase concentration and reduce ORP resulting in an increase of methane production. At a CoFe2O4 concentration of 200 mg/L, the cumulative methane production reached to 414.75 mL/(gVS) which was increased by 79.1% compared to the control group(231.59 mL/(gVS)). Meanwhile, the variation of microbial community showed that CoFe2O4 could increase the relative abundance of Methanobacterium and the methane production pathway in the AD was shifted from acetoclastic to hydrogenotrophic.

Microbial community succession and antibiotic resistance gene response during the one-step startup of thermophilic UASB

A thermophilic anaerobic up-flow anaerobic sludge bed (UASB) was rapidly established from a mesophilic reactor in 25 days by directly increasing the operation temperature from 35 to 55℃. The succession of methanogenic microbial community and response of antibiotic resistance genes (ARGs) during the startup period were revealed by 16S rRNA gene amplicon sequencing and metagenomics analysis. After the temperature was increased, thermophilic hydrotropic Methanothermobacter outgrew aceticlastic Methanosaeta and the mesophilic hydrotropic methanogens (Methanobacterium and Methanolinea), with its abundance increasing from 3.19 % on day 7 to 72.33 % on day 10. Meanwhile, Exilispira, an acetate-oxidizing bacterium, proliferated quickly in the fermenter community concurrently with the growth of Methanothermobacter, indicating that the syntrophic relationship between acetate-oxidizing organisms and hydrogenotrophic methanogens is the primary pathway for methane production in thermophilic UASB. As for ARGs, the abundance of the resistome of the anaerobic sludge decreased after the hike in temperature, driven by the succession of the methanogenic community, confirming the benefit of thermophilic treatment for antimicrobial resistance control. The results proved that a thermophilic UASB could be rapidly constructed by a one-step startup strategy due to the fast establishment of the syntrophic acetate oxidation pathway, and maintain better ARG control performance than its mesophilic counterpart.

Performance evaluation and microbial community analysis of a continuous stirred tank reactor–anaerobic ceramic membrane bioreactor system for practical swine wastewater treatment

To treat swine wastewater from a practical piggery, a continuous stirred tank reactor (CSTR)–anaerobic ceramic membrane bioreactor (AnCMBR) was operated for 201 days. A high chemical oxygen demand (COD) removal efficiency of 95 % and biogas production rate of 0.31 L/L·d were achieved at an organic loading rate (OLR) of 1.19 g-COD/L·d. Although the free ammonia concentration reached a maximum of 200 mg/L during the last operational phase, no evident inhibitory effect on AnCMBR performance was observed. Microbial community analyses indicated that six bacterial phyla were responsible for the hydrolysis, acidification, and fermentation of organics and environmental stabilization. Hydrotrophic methanogenic archaea were dominant, indicating that the main methane production pathway involved the use of H2 and CO2 as substrates. The results demonstrated that CSTR–AnCMBR systems exhibit excellent organics removal performance and energy recovery at high organic concentrations and possess strong anti-shock ability. Thus, they represent alternative green and sustainable anaerobic technologies for practical swine wastewater treatment.

Optimization of the anaerobic digestion process and the metabolic pathway of methanogenesis of Jerusalem artichoke straw with the mixed addition of iron filings and biochar

This study explored the feasibility of the mixed addition of iron filings and biochar in the anaerobic digestion of Jerusalem artichoke straw (JAS), using iron filings and biochar as mixed additives for mid-temperature batch anaerobic digestion experiments, and applied response surface methodology to investigate the interactive effects of different ratios of iron filings and biochar (FE/BC), solid content, and initial pH on the cumulative methane production from JAS. The results showed that the optimal conditions for methane production were an FE/BC of 2.45:7.55, an initial pH of 7.59, and a solid content of 8.80 %. Under these conditions, the cumulative methane production was 278.11 mL g−1 volatile solid (VS) contents, which was very close to the predicted value (286.42 mL g−1 VS), with a relative error of less than 5 %, indicating the effectiveness of the model. The optimized conditions with mixed addition (HB) exhibited good synergistic promotion, increasing cumulative methane production by 48.69 % compared to the control without additives and by 22.29 % and 29.57 % compared to the single addition of iron filings and biochar, respectively. Moreover, the HB treatment under optimized conditions achieved good economic benefits. Metagenomic analysis showed that the relative abundances of bacterial taxa including Fermentimonas, DMER64, Bacteroidetes_VadinHA17, unclassified Synergistaceae, Clostridium, Ruminiclostridium, and unclassified Anaerolineaceae were higher in HB samples, and Methanothrix was also enriched. Methane metabolism pathway analysis indicated that the anaerobic digestion of JAS with the mixed addition of iron filings and biochar enhanced the acetoclastic methane production pathway, thereby increasing methane production.

High concentrate diets altered the structure and function of rumen microbiome in goats.

This study used metatranscriptomics to investigate the effects of concentrate diet level on rumen microbiome composition and function in goats. A total of 12 healthy 120-day-old Da'er goats were randomly allotted into two treatments: L group (low dietary concentrate level group, concentrate: forage ratio was 25: 75) and H group (high dietary concentrate level group, concentrate: forage ratio was 80: 20). The study included a 10-day pre-feeding period and a 60-day growth experiment. The results showed that compared with the L group, the average daily gain and the slaughter rate in the H group were increased, while the F/G was decreased; the concentration of lactate and ammonia nitrogen, and the proportion of butyrate and valerate in the rumen of the H group were increased, while the proportion of acetate, and the ratio of acetate to propionate were decreased (p < 0.05). Among rumen bacteria, compared with the L group, the H group significantly decreased the relative abundance of Firmicutes and Fibrobacteria at the phylum level, decreased the relative abundance of Bacteroidetes, Fibrobacter, and Sarcina and increased the relative abundance of Clostridium at the genus level, and decreased the relative abundance of Fibrobacter succinogenes, Sarcina sp. DSM 11001, Oscillibacter sp. KLE 1728, and Ruminococcus flavefaciens and increased the relative abundance of Clostridium sp. ND2 and Firmicutes bacteria CAG: 103 at the species level (p < 0.05). Among rumen fungi, the relative abundance of Basidiomycota, Neocallimastigomycota, Mortierella, Mortierella elongata, and Gonapodyna prolifera was lower in the H group than that in the L group (p < 0.05). Functional annotation results showed that the abundance of Glycoside hydrolases genes in rumen microbiome was significantly decreased in the H group compared to the L group (p < 0.05). The result of KEGG DEGs enrichment analysis showed that the gene expression of cellulose 1,4-β-cellobiosidase, acetyl-CoA hydrolase, lactate dehydrogenase, succinate-semialdehyde dehydrogenase, D-malate dehydrogenase and related genes in methane production pathways of rumen microbiome was decreased in the H group. In summary, feeding high concentrate diets improved the production performance of goats, altered the structure and composition of rumen microbiome and changed the function of rumen microbiome.

Microbial community changes and metabolic pathways analysis during waste activated sludge and meat processing waste anaerobic co-digestion

Waste activated sludge (WAS) and meat processing waste (MPW) were acted as co-substrates in anaerobic co-digestion (AcD), and biochemical methane potential (BMP) test was carried out to investigate the methane production performances. Microbial community structure and metabolic pathways analyses were conducted by 16S rRNA high-throughput sequencing and functional prediction analysis. BMP test results indicated that AcD of 70% WAS+30% MPW and 50% WAS+50% MPW (VS/VS) could significantly improve methane yield to 371.05 mL/g VS and 599.61 mL/g VS, respectively, compared with WAS acting as sole substrate (191.87 mL/g VS). The results of microbial community analysis showed that Syntrophomonas and Petrimonas became the dominant bacteria genera, and Methanomassiliicoccus and Methanobacterium became the dominant archaea genera after MPW addition. 16S functional prediction analysis results indicated that genes expression of key enzymes involved in syntrophic acetate oxidation (SAO), hydrogenotrophic and methylotrophic methanogenesis were up-regulated, and acetoclastic methanogenesis was inhibited after MPW addition. Based on these analyses, it could be inferred that SAO combined with hydrogenotrophic and methylotrophic methanogenesis was the dominant pathway for organics degradation and methane production during AcD. These findings provided systematic insights into the microbial community changes and metabolic pathways during AcD of WAS and MPW.

Riboflavin modified carbon cloth enhances anaerobic digestion treating food waste in a pilot-scale system.

Previous laboratory-scale studies have consistently shown that carbon-based conductive materials can notably improve the anaerobic digestion of food waste, typically employing reactors with regular capacity of 1-20L. Furthermore, incorporating riboflavin-loaded conductive materials can further address the imbalance between fermentation and methanogenesis in anaerobic systems. However, there have been few reports on pilot-scale investigation. In this study, a 10m2 of riboflavin modified carbon cloth was incorporated into a pilot-scale (2m3) food waste anaerobic reactor to improve its treatment efficiency. The study found that the addition of riboflavin-loaded carbon cloth can increase the maximum organic loading rate (OLR) by 40% of the pilot-scale reactor, compared to the system using carbon cloth without riboflavin loading, while ensuring efficient operation of the reaction system, effectively alleviating system acidification, sustaining methanogen activity, and increasing daily methane production by 25%. Analysis of the microbial community structure revealed that riboflavin-loaded carbon cloth enriched the methanogenic archaea in the genera of Methanothrix and Methanobacterium, which are capable of extracellular direct interspecies electron transfer (DIET). And metabolic pathway analysis identified the methane production pathway, highly enriched on the reduction of acetic acid and CO2 at riboflavin-loaded carbon cloth sample. The expression levels of genes related to methane production via DIET pathway were also significantly upregulated. These results can provide important guidance for the practical application of food waste anaerobic digestion engineering.

Effects of different nitrogen applications and straw return depth on straw microbial and carbon and nitrogen cycles in paddy fields in the cool zone

Straw is an important source of organic fertilizer for soil enrichment, however, the effects of different nitrogen(N) application rates and depths on straw decomposition microorganisms and carbon and nitrogen cycling under full straw return conditions in cool regions of Northeast China are not clear at this stage. In this paper, we applied macro-genome sequencing technology to investigate the effects of different N application rates (110 kg hm−2, 120 kg hm−2, 130 kg hm−2, 140 kg hm−2, 150 kg hm−2) and depths (0–15 cm, 15–30 cm) on straw decomposing microorganisms and N cycling in paddy fields in the cool zone of Northeast China. The results showed that (1) about 150 functional genes are involved in the carbon cycle process of degradation during the degradation of returned straw, of which the largest number of functional genes are involved in the methane production pathway, about 42, the highest abundance of functional genes involved in the citric acid cycle pathway. There are 22 kinds of functional genes involved in the nitrogen cycle degradation process, among which there are more kinds involved in nitrogen fixation, with 4 kinds. (2) High nitrogen application (150 kg hm−2) inhibited the carbon and nitrogen conversion processes, and the abundance of straw-degrading microorganisms and nitrogen-cycling functional genes was relatively high at a nitrogen application rate of 130 kg hm−2. (3) Depth-dependent heterogeneity of the microbial community was reduced throughout the vertical space. At 71 days of straw return, the nitrogen cycling function decreased and some carbon functional genes showed an increasing trend with the increase of straw return depth. The nitrogen cycle function decreased with the increase of straw returning depth. The microbial community structure was best and the abundance of functional genes involved in the nitrogen cycling process was higher under the conditions of 0–15 cm of returning depth and 130 kg hm−2 of nitrogen application.

Anaerobic methane oxidation is quantitatively important in deeper peat layers of boreal peatlands: Evidence from anaerobic incubations, in situ stable isotopes depth profiles, and microbial communities

Boreal peatlands store most of their carbon in layers deeper than 0.5 m under anaerobic conditions, where carbon dioxide and methane are produced as terminal products of organic matter degradation. Since the global warming potential of methane is much greater than that of carbon dioxide, the balance between the production rates of these gases is important for future climate predictions. Herein, we aimed to understand whether anaerobic methane oxidation (AMO) could explain the high CO2/CH4 anaerobic production ratios that are widely observed for the deeper peat layers of boreal peatlands. Furthermore, we quantified the metabolic pathways of methanogenesis to examine whether hydrogenotrophic methanogenesis is a dominant methane production pathway for the presumably recalcitrant deeper peat. To assess the CH4 cycling in deeper peat, we combined laboratory anaerobic incubations with a pathway-specific inhibitor, in situ depth patterns of stable isotopes in CH4, and 16S rRNA gene amplicon sequencing for three representative boreal peatlands in Western Siberia. We found up to a 69 % reduction in CH4 production due to AMO, which largely explained the high CO2/CH4 anaerobic production ratios and the in situ depth-related patterns of δ13C and δD in methane. The absence of acetate accumulation after inhibiting acetotrophic methanogenesis and the presence of sulfate- and nitrate-reducing anaerobic acetate oxidizers in the deeper peat indicated that these microorganisms use SO42− and NO3− as electron acceptors. Acetotrophic methanogenesis dominated net CH4 production in the deeper peat, accounting for 81 ± 13 %. Overall, anaerobic oxidation is quantitatively important for the methane cycle in the deeper layers of boreal peatlands, affecting both methane and its main precursor concentrations.

Enhancement of hydrogenotrophic methanogenesis for methane production by nano zero-valent iron in soils

Nanoscale zero-valent iron (nZVI) is attracting increasing attention as the most commonly used environmental remediation material. However, given the high surface area and strong reducing capabilities of nZVI, there is a lack of understanding regarding its effects on the complex anaerobic methane production process in flooded soils. To elucidate the mechanism of CH4 production in soil exposed to nZVI, paddy soil was collected and subjected to anaerobic culture under continuous flooding conditions, with various dosages of nZVI applied. The results showed that the introduction of nZVI into anaerobic flooded rice paddy systems promoted microbial utilization of acetate and carbon dioxide as carbon sources for methane production, ultimately leading to increased methane production. Following the introduction of nZVI into the soil, there was a rapid increase in hydrogen levels in the headspace, surpassing that of the control group. The hydrogen levels in both the experimental and control groups were depleted by the 29th day of culture. These findings suggest that nZVI exposure facilitates the enrichment of hydrogenotrophic methanogens, providing them with a favorable environment for growth. Additionally, it affected soil physicochemical properties by increasing pH and electrical conductivity. The metagenomic analysis further indicates that under exposure to nZVI, hydrogenotrophic methanogens, particularly Methanobacteriaceae and Methanocellaceae, were enriched. The relative abundance of genes such as mcrA and mcrB associated with methane production was increased. This study provides important theoretical insights into the response of key microbes, functional genes, and methane production pathways to nZVI during anaerobic methane production in rice paddy soils, offering fundamental insights into the long-term fate and risks associated with the introduction of nZVI into soils.

Can meta-omics revolutionize our understanding of rumen methane emissions?

Abstract The global challenge of methane emissions from enteric fermentation is critical, as it contributes significantly to atmospheric greenhouse gases and represents a loss of energy that could otherwise be utilized by ruminants. With the increasing demand for dairy and meat products, finding effective methods to reduce methane production is essential. This review explores the use of advanced meta-omics techniques – including metagenomics, metatranscriptomics, metaproteomics, and metabolomics – to deepen our understanding of ruminal methane production and identify potential strategies for its mitigation. These high-throughput technologies provide comprehensive insights into the rumen microbial communities and their metabolic functions by analyzing DNA, RNA, proteins, and metabolites directly from environmental samples. Metagenomics and metatranscriptomics offer a detailed view of microbial diversity and gene expression, while metaproteomics can identify specific enzymes and proteins directly involved in methane production pathways, revealing potential targets for mitigation strategies. Integrating these meta-omics approaches allows for a holistic understanding of the microbial processes that drive methane emissions, enabling the development of more precise interventions, such as tailored dietary modifications and the use of specific inhibitors. This review underscores the importance of a multi-omics strategy in characterizing microbial roles and interactions within the rumen, which is crucial for devising effective and sustainable methods to reduce methane emissions without compromising livestock productivity.

Generation of secondary microbial methane of high-rank coals: insights from the microbial community and carbon isotope.

Secondary microbial methane could provide a valuable energy source if it were better understood. Although coal seam is an ideal environment for investigating secondary microbial methane, there are few studies to trace the secondary microbial methane of high-rank coals. Here, we collected co-produced water samples from coalbeds in the Qinshui Basin (China) and analyzed the microbial community structure by 16S ribosomal RNA (16S rRNA) amplicon sequencing analysis. 16S rRNA sequencing demonstrated abundant methanogens in coalbeds including 6 orders (Methanobacteriales, Methanococcales, Methanofastidiosales, Methanomassiliicoccale, Methanomicrobiales, and Methanosarciniales) and 22 genera of methanogens. Superheavy DIC (δ13CDIC ranging from -4.2‰ to 34.8‰) and abundance of methanogenic microbes in co-produced water revealed the generation of secondary biogenic methane in high-rank coal seams in the Qingshui Basin. Hydrogenotrophic methanogenesis is the main pathway for secondary biogenic methane production. In deeply buried coal seams, biogenic methane is dominated by CO2 and H2 reduction methanogenesis, and in shallow buried coal seams, it may be produced synergistically by hydrocarbon degradation and hydrogenotrophic methanogenic microbes. The study discussed here is important for a better understanding of the generation of secondary microbial methane in high-rank coal.

Microbial adaptation to high ammonia environment in submerged anaerobic membrane bioreactor under volatile fatty acids and ammonia stresses

Anaerobic membrane bioreactor (AnMBR) is a high-rate bioreactor which is preferred over other configurations due to its potential to completely decouple solid retention time from hydraulic retention time. However, limited information is available on the bioreactor behavior under stressed environment, particularly during the startup period. The objective of this study was to investigate the effects of ammonium (NH4+) and organic loading rate (OLR) on the digestion performance and microbial population of submerged AnMBR. The findings revealed that high NH4+ was positively correlated with high methane concentration while high OLR increased methane production. The microbial analysis showed that both bacteria and archaea were influenced by the change of NH4+ and OLR. Particularly, at high OLR, Bacteroidetes were shown to be more sensitive to high NH4+ than Firmicutes, and Chloroflexi tolerated NH4+ even up to 2.5 g/L. Moreover, NH4+ was possibly the first driving factor in the change of methane production pathways from acetoclastic methanogenesis to hydrogenotrophic and methylotrophic methanogenesis. Membrane biofouling was also caused mainly by bacterial community (Firmicutes and Chloroflexi), while the contribution of archaeal community was minimal (Methanosaetaceae).

Responses of methane production and methanogenic pathways to polystyrene nanoplastics exposure in paddy soil

Nanoplastics (NPs) have attracted increasing attention within terrestrial ecosystems. However, our understanding of their impacts on the intricate anaerobic methanogenesis processes occurring in paddy soils microbial communities remains limited with respect to nanoplastics shape, function, and metabolic effects. Herein, we explored the effects of polystyrene nanoplastics (PS-NPs) and microplastics (PS-MPs) on anaerobic methanogenesis in a typical paddy soil. The results show that PS-NPs delayed methane production and the time to reach peak acetate content in incubation process of paddy soils, and the methanogenic rate increased rapidly after 13 days, with a maximum increase of 87.97%. However, PS-MPs had no marked effect on CH4, CO2 and acetate production. In addition, PS-NPs affected soil physicochemical properties by reducing pH and increasing electrical conductivity. Acetoclastic methanogens were enriched and the relative abundance of the genes ackA, pta, ACSS, cdhC, cdhD and cdhE in the acetoclastic pathways were significantly increased under PS-NPs exposure. In addition, PS-MPs had significant effect on the microbial community structure but no effect on methanogenic pathways of the paddy soils. This study provides important insights into the response of key microorganisms, functional genes and methanogenesis pathways to NPs during anaerobic methanogenesis in paddy soils.

Environmental impact assessment of a novel third-generation biorefinery approach for astaxanthin and biofuel production

A novel third-generation biorefinery approach, including two paths of Ethanol/methane production pathway (EMP) and the direct methane production pathway (DMP), for astaxanthin and ethanol and biogas production from the freshwater microalgae Haematococcus pluvialis was developed previously. To ensure its environmental sustainability, a comprehensive life cycle assessment (LCA) study was conducted based on 1-GJ energy generation from biomethane as the functional unit. Results indicate that the EMP pathway had higher environmental impacts on all categories due to more stages and chemicals/energy consumption (at least five times greater effect). Results showed that while the enzymatic hydrolysis step followed by the fermentation stage was the main contributor to all environmental categories in the EMP route, astaxanthin induction dominated all environmental categories in the DMP route. The results showed that sodium nitrate, phosphate salts, inoculum sludge, acetone, and electricity had considerable environmental impacts. Moreover, despite low enzyme usage in enzymatic hydrolysis, these proteins significantly impacted all environmental categories in this stage. The baseline analysis concluded that to produce 1 GJ energy from methane, about 88 kg and 13 kg CO2 were generated from the EMP and DMP pathways, respectively. A sensitivity analysis was also conducted to compare various ratios of chemicals, such as phosphate salts, with high contributions to enzymatic hydrolysis and astaxanthin induction stages in the EMP and DMP routes, respectively. Finally, the LCA results revealed that the DMP pathway is more environmentally friendly with the same economic value of biomethane and astaxanthin production. This LCA study updated the data related to the environmental assessment of processes to utilize H. pluvialis to produce biofuels and astaxanthin simultaneously.