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Oxic methane production (OMP) has been reported to significantly contribute to methane emissions from oxic surface waters. Demethylation of organic compounds, photosynthesis-associated methane production, and (bacterio)chlorophyll reduction activity are some of the investigated mechanisms as potential OMP sources related to photosynthetic organisms. Recently, cyanobacteria have often been correlated with methane accumulation and emission in freshwater, marine, and saline systems. The Brazilian Pantanal is the world's largest wetland system, with approximately 10,000 shallow lakes, most of which are highly alkaline and saline extreme environments. We initiated this study with an overall investigation using genetic markers, from which we explored metagenomic and limnological data from the Pantanal soda for five potential OMP pathways. Our results showed a strong positive correlation between dissolved methane concentrations and bloom events. Metagenomic data and nutrients, mainly orthophosphate, nitrogen, iron, and methane concentrations, suggest that the organic phosphorous demethylation pathway has the most potential to drive OMP in lakes with blooms. A specialized bacterial community was identified, including the Cyanobacteria Raphidiopsis, although the bloom does not contain the genes to carry out this process. These data showed enough evidence to infer the occurrence of an OMP pathway at Pantanal soda lakes, including the microbial sources and their relation to the cyanobacterial blooms.

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The anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) has shown to be a viable alternative since it allows energy recovery in the form of methane and generates a residue (digestate) that can be applied effectively as a soil improver or fertiliser. The potential for methane production and the digestates' characteristics depend on the substrate characteristics and the process variables such as temperature, solids retention time, and organic load. This study dealt with OFMSW anaerobic digestion under different organic loading rates and temperatures and the characteristics of the resulting digestates. Three semi-continuous reactors were operated at 20, 35, and 55°C and fed daily with ground, fresh OFMSW from Mexico City. The inoculum was temperature-adapted UASB granular sludge. The main results indicate that the anaerobic digestion was adequate, as the pH values were slightly alkaline, which is sufficient for methanization, and the alkalinity was not a limiting factor. Potassium and PO4-P increased with the organic load, and Kjeldahl nitrogen decreased. At 20°C, total organic carbon (TOC) increased substantially with the organic load; at 35°C, it remained without significant changes; and at 55°C, TOC slightly decreased with the organic load. The C/N ratio changed accordingly to TOC variations. At 20°C, the residual biogas potential increased with the organic load; at 35 and 55°C, it decreased with increasing organic load; the residual biogas potential increased with residual fatty acids concentrations. To comply with international standards for agricultural use, the digestates need only dewatering and supplementing with PO4-P.

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OBJECTIVES: This study evaluated the effect of particle size and dosage of granular activated carbon (GAC) on methane production from the anaerobic digestion of raw effluent (RE) of swine wastewater, and the solid (SF) and liquid (LF) fractions. The effect of temperature using the selected size and dosage of GAC was also evaluated. METHODS: 60 mL of swine wastewater were inoculated with anaerobic granular sludge and GAC at different dosages and particle size. The cultures were incubated at different temperatures at 130 rpm. The kinetic parameters from experimental data were obtained using the Gompertz model. RESULTS: The cultures with the LF and GAC (75-150 µm, 15 g/L) increased 1.87-fold the methane production compared to the control without GAC. The GAC at 75-150 µm showed lower lag phases and higher Rmax than the cultures with GAC at 590-600 µm. The cumulative methane production at 45 °C with the RE + GAC was 7.4-fold higher than the control. Moreover, methane production at 45 °C significantly increased with the cultures LF + GAC (6.0-fold) and SF + GAC (2.0-fold). The highest production of volatile fatty acids and ammonium was obtained at 45 °C regardless of the substrate and the addition of GAC contributed to a higher extent than the cultures lacking GAC. In most cases, the kinetic parameters at 30 °C and 37 °C were also higher with GAC. CONCLUSIONS: GAC contributed to improving the fermentative and methanogenesis stages during the anaerobic digestion of fractions, evidenced by an improvement in the kinetic parameters.

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Anaerobic digestion (AD) of swine waste allows obtaining renewable energy, biofertilizer and the reduction of environmental impacts. However, the low C:N ratio of pig manure generates high concentrations of ammonia nitrogen in the digestion process, reducing methane production. Zeolite is an effective ammonia adsorbent; thus, in this research the ammonia adsorption capacity of natural Ecuadorian zeolite was studied under different operating conditions. Subsequently, its effect on methane production from swine waste was evaluated using three doses of zeolite, 1.0, 4.0 and 8.0 g, in 1 L batch bioreactors. The results showed that the Ecuadorian natural zeolite has an adsorption capacity of around 19 mgNH3-N gZ-1 when using ammonium chloride solution and, an adsorption capacity between 37 and 65 mgNH3-N gZ-1 using swine waste. On the other hand, the addition of zeolite had a significant effect on methane production (p < 0.01). The zeolite doses that provided the highest methane production were 4.0 and 8.0 g L-1, which led to values of 0.375 and 0.365 Nm3CH4 kgVS-1, compared to the values of 0.350 and 0.343 Nm3CH4 kgVS-1 that were obtained for the treatments without addition of zeolite and using a dose of 1.0 g L-1, respectively. Addition of natural Ecuadorian zeolite meant not only a significant increase on methane production in the AD of swine waste, but also a better quality of the biogas with higher percentages of methane and lower concentrations of H2S.

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Anaerobic co-digestion (AcoD) of sugarcane vinasse and glycerol can be profitable because of the destination of two biofuel wastes produced in large quantities in Brazil (ethanol and biodiesel, respectively) and the complementary properties of these substrates. Thus, the objective of this study was to assess the effect of increasing the organic loading rate (OLR) from 2 to 20 kg COD m-3 d-1 on the AcoD of vinasse and glycerol (50 %:50 % on a COD basis) in a thermophilic (55 °C) anaerobic fluidized bed reactor (AFBR). The highest methane production rate was observed at 20 kg COD m-3 d-1 (8.83 L CH4 d-1 L-1), while the methane yield remained stable at around 265 NmL CH4 g-1 CODrem in all conditions, even when influent vinasse reached 1811 mg SO42- L-1 (10 kg COD m-3 d-1). Sulfate was not detected in the effluent. Bacterial genera related to sulfate removal, such as Desulfovibrio and Desulfomicrobium, were observed by means of shotgun metagenomic sequencing at 10 kg COD m-3 d-1, as well as the acetoclastic archaea Methanosaeta and prevalence of genes encoding enzymes related to acetoclastic methanogenesis. It was concluded that process efficiency and methane production occurred even in higher sulfate concentrations due to glycerol addition.

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Background: Ruminants harbor a complex microbial community within their gastrointestinal tract, which plays major roles in their health and physiology. Brazil is one of the largest producers of beef in the world and more than 90% of the beef cattle herds are composed of pure and crossbred Nelore (Bos indicus). Despite its importance to the Brazilian economy and human feeding, few studies have characterized the Nelore microbiome. Therefore, using shotgun metagenomics, we investigated the impact of diet on the composition and functionality of the Nelore microbiome, and explored the associations between specific microbial taxa and their functionality with feed efficiency and methane emission. Results: The ruminal microbiome exhibited significantly higher microbial diversity, distinctive taxonomic profile and variations in microbial functionality compared to the fecal microbiome, highlighting the distinct contributions of the microbiomes of these environments. Animals subjected to different dietary treatments exhibited significant differences in their microbiomes' archaeal diversity and in the abundance of 89 genera, as well as in the functions associated with the metabolism of components of each diet. Moreover, depending on the diet, feed-efficient animals and low methane emitters displayed higher microbial diversity in their fecal microbiome. Multiple genera were associated with an increase or decrease of the phenotypes. Upon analyzing the functions attributed to these taxa, we observed significant differences on the ruminal taxa associated with feed efficient and inefficient cattle. The ruminal taxa that characterized feed efficient cattle stood out for having significantly more functions related to carbohydrate metabolism, such as monosaccharides, di-/oligosaccharides and amino acids. The taxa associated with methane emission had functions associated with methanogenesis and the production of substrates that may influence methane production, such as hydrogen and formate. Conclusion: Our findings highlight the significant role of diet in shaping Nelore microbiomes and how its composition and functionality may affect production traits such as feed efficiency and methane emission. These insights provide valuable support for the implementation of novel feeding and biotechnological strategies.

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A challenge in the study of gastrointestinal microbiota (GITm) is the validation of the genomic data with metabolic studies of the microbial communities to understand how the microbial networks work during health and sickness. To gain insights into the metabolism of the GITm, feces from healthy and sick rats with cancer were inoculated in a defined synthetic medium directed for anaerobic prokaryote growth (INC-07 medium). Significant differences between cultures of healthy and sick individuals were found: 1) the consumption of the carbon source and the enzyme activity involved in their catabolism (e.g., sucrase, lactase, lipases, aminotransferases, and dehydrogenases); 2) higher excretion of acetic, propionic, isobutyric, butyric, valeric, and isovaleric acids; 3) methane production; 4) ability to form biofilms; and 5) up to 500 amplicon sequencing variants (ASVs) identified showed different diversity and abundance. Moreover, the bowel inflammation induced by cancer triggered oxidative stress, which correlated with deficient antioxidant machinery (e.g., NADPH-producing enzymes) determined in the GITm cultures from sick individuals in comparison with those from control individuals. Altogether, the data suggested that to preserve the microbial network between bacteria and methanogenic archaea, a complete oxidation of the carbon source may be essential for healthy microbiota. The correlation of 16S rRNA gene metabarcoding between cultures and feces, as well as metabolomic data found in cultures, suggest that INC-07 medium may be a useful tool to understand the metabolism of microbiota under gut conditions.

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Carbon-based materials have been shown to enhance anaerobic digestion processes by promoting direct interspecies electron transfer in methanogenic consortia. However, little is known on their effects during the treatment of complex substrates, such as those derived from protein-rich wastewaters. Here, organic xerogels (OX) are tested, for the first time, as accelerators of the methanogenic activity of an anaerobic consortium treating a synthetic protein-rich wastewater. Three OX with distinct pore size distribution (10 and 1000 nm for OX-10 and OX-1000, respectively) and structural conformation (graphene oxide integration into OX-10-GO polymeric matrix) were synthesized. OX-1000 promoted the highest methane production rate (5.21 mL/g*h, 13.5% increase with respect to the control incubated without OX) among the synthesized OX. Additionally, batch bioreactors amended with OX achieved higher chemical oxygen demand (COD) removal (up to 88%) as compared to the control, which only showed 50% of COD removal. Interestingly, amendment of bioreactors with OX also triggered the production of medium-chain fatty acids, including caprylate and caproate. Moreover, OX decreased the accumulation of ammonium, derived from proteins hydrolysis, partly explained by their adsorption capacities, and probably involving their electron-accepting capacity promoting anaerobic ammonium oxidation. This is the first time that OX were successfully applied as methanogenic accelerators for the anaerobic treatment of synthetic protein-rich wastewater, increasing the methane production rate and COD removal as well as triggering the production of medium chain fatty acids and attenuating the accumulation of ammonium. Therefore, OX are proposed as suitable materials to boost the efficiency of anaerobic systems to treat complex industrial wastewaters.

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The aim of this study was to evaluate the theoretical potential of methane production of the food waste generated by a university restaurant, as well as to verify the influence of the fruit and vegetable waste in the feeding composition of an anaerobic bioreactor treating this type of waste. Four feeding compositions combining three fractions of the food waste (fruit and vegetable fraction, soy protein and beans fraction, and rice fraction) at different concentrations were tested in anaerobic processes lasting 10 and 30 days. Additionally, a study of the theoretical potential of methane production from each fraction that composes the food waste was carried out, as well as the evaluation of the specific methanogenic activity of the anaerobic sludge. Despite its low theoretical potential of methane production (0.037 LCH4/g), the presence of the fruit and vegetable mixture in three of the feeding compositions led to greater organic matter degradation (above 69%) and CH4 yields (above 0.20 LCH4/gVS) in both periods tested, in comparison with the achieved by the feeding composition lacking this fraction. The results suggest that the presence of the fruit and vegetable mixture contributed with the supplementation of micro- and macroelements to the anaerobic sludge during the digestion of food waste.

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El Chichón volcano is one of the most active volcanoes in Mexico. Previous studies have described its poly-extreme conditions and its bacterial composition, although the functional features of the complete microbiome have not been characterized yet. By using metabarcoding analysis, metagenomics, metabolomics and enzymology techniques, the microbiome of the crater lake was characterized in this study. New information is provided on the taxonomic and functional diversity of the representative Archaea phyla, Crenarchaeota and Euryarchaeota, as well as those that are representative of Bacteria, Thermotogales and Aquificae. With culture of microbial consortia and with the genetic information collected from the natural environment sampling, metabolic interactions were identified between prokaryotes, which can withstand multiple extreme conditions. The existence of a close relationship between the biogeochemical cycles of carbon and sulfur in an active volcano has been proposed, while the relationship in the energy metabolism of thermoacidophilic bacteria and archaea in this multi-extreme environment was biochemically revealed for the first time. These findings contribute towards understanding microbial metabolism under extreme conditions, and provide potential knowledge pertaining to "microbial dark matter", which can be applied to biotechnological processes and evolutionary studies.

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