Enhanced thermophilic high-solids anaerobic digestion of organic fraction of municipal solid waste with spatial separation from conductive materials in a single reactor volume.

Despite benefits such as lower water and working volume requirements, thermophilic high solids anaerobic digestion (THSAD) often fails due to the rapid build-up of volatile fatty acids (VFAs) and the associated drop in pH. Use of conductive materials (CM) can promote THSAD through stimulation of direct interspecies electron transfer (DIET), while the need for their constant dosing due to poor separation from effluent impairs economic feasibility. This study used an approach of spatially separating magnetite and granular activated carbon (GAC) from the organic fraction of municipal solid waste (OFMSW) in a single reactor for THSAD. GAC and magnetite addition could both mitigate the severe inhibition of methanogenesis after VFAs build-up to ∼28-30 g/L, while negligible methane production was observed in the control group. The highest methane yield (286 mL CH4/g volatile solids (VS)) was achieved in magnetite-added reactors, while the highest maximum CH4 production rates (26.38 mL CH4/g VS/d) and lowest lag-phase (2.83 days) were obtained in GAC-added reactors. The enrichment of GAC and magnetite biofilms with various syntrophic and potentially electroactive microbial groups (Ruminiclostridium 1, Clostridia MBA03, Defluviitoga, Lentimicrobiaceae) in different relative abundances indicates the existence of specific preferences of these groups for the nature of CM. According to predicted basic metabolic functions, CM can enhance cellular processes and signals, lipid transport and metabolism, and methane metabolism, resulting in improved methane production. Rearrangement of metabolic pathways, formation of pili-like structures, enrichment of biofilms with electroactive groups and a significant improvement in THSAD performance was attributed to the enhancement of the DIET pathway. Promising results obtained in this work due to the spatial separation of the bulk OFMSW and CM can be useful for modeling larger-scale THSAD systems with better recovery of CM and cost-effectiveness.

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.

Utilization of agricultural residues for energy and resource recovery towards a sustainable environment.

Fungal pre-treatment using Pleurotus ostreatus (PO) was carried out on individual and combinations of agro-waste wheat straw (WS), rice straw (RS), and pearl millet straw (PMS) with the addition of biochar (5%,7.5% and 10%) to reduce the pre-treatment duration. Further remaining substrate known as spent mushroom substrate (SMS) was used in anaerobic digestor (AD) for estimation enhanced biomethane yield. Equal ratios of RS + WS, WS + PMS, PMS + RS, and RS + PMS + WS and biochar addition were taken for enhancing pre-treatment, PO growth and AD process. The extent of pre-treatment was recorded with the maximum lignin removal of 40.4% for RS + PMS + WS as compared to untreated counterparts and 0.5%, 2.2%, and 3.3% times more lignin removal from individual PMS, RS, and WS respectively. Addition of biochar to the substrates reduced the total pre-treatment duration by days as compared to the non-biochar substrates. Biological efficiency (BE) used for the analysis of mushroom growth varied from 51-92%. Further, the average bio-methane yield was 187 ml/gVS for SMS of PMS + WS + RS with 10% biochar indicating an increment of 83.33% from untreated SMS of PMS + WS + RS. This, higher biomethane yield was 9.35%, 22.22% and 57.14% times higher than individual SMS of PMS, RS, and WS respectively. The current study shows that biochar not only enhances the bio-methane yield but also reduces the biological pre-treatment duration and removes the dependency on one lignocellulosic biomass for energy (bio-methane) and food (mushroom) production.

Methane in West Siberia terrestrial seeps: Origin, transport, and metabolic pathways of production.

The expansive plains of West Siberia contain globally significant carbon stocks, with Earth's most extensive peatland complex overlying the world's largest-known hydrocarbon basin. Numerous terrestrial methane seeps have recently been discovered on this landscape, located along the floodplains of the Ob and Irtysh Rivers in hotspots covering more than 2500 km2 . We articulated three hypotheses to explain the origin and migration pathways of methane within these seeps: (H1) uplift of Cretaceous-aged methane from deep petroleum reservoirs along faults and fractures, (H2) release of Oligocene-aged methane capped or trapped by degrading permafrost, and (H3) horizontal migration of Holocene-aged methane from surrounding peatlands. We tested these hypotheses using a range of geochemical tools on gas and water samples extracted from seeps, peatlands, and aquifers across the 120,000 km2 study area. Seep-gas composition, radiocarbon age, and stable isotope fingerprints favor the peatland hypothesis of seep-methane origin (H3). Organic matter in raised bogs is the primary source of seep methane, but observed variability in stable isotope composition and concentration suggest production in two divergent biogeochemical settings that support distinct metabolic pathways of methanogenesis. Comparison of these parameters in raised bogs and seeps indicates that the first is bogs, via CO2 reduction methanogenesis. The second setting is likely groundwater, where dissolved organic carbon from bogs is degraded via chemolithotrophic acetogenesis followed by acetate fermentation methanogenesis. Our findings highlight the importance of methane lateral migration in West Siberia's bog-dominated landscapes via intimate groundwater connections. The same phenomenon could occur in similar landscapes across the boreal-taiga biome, thereby making groundwater-fed rivers and springs potent methane sources.

Insights into sustainable resource and energy recovery from leachate towards emission mitigation for environmental management: A critical approach.

The exponential generation of municipal solid waste (MSW) and landfill disposal without any treatment has increased the continuous generation of landfill leachate. Improper MSW and leachate management are contributing to environmental degradation and water and soil pollution, which must be treated. Numerous works have been conducted on leachate treatments for energy and resource recovery. This review presents a comprehensive study of leachate management in which different treatment methods are discussed to analyze the suitability of processes that can be employed to treat leachate efficiently. Further, the characteristics of leachate are examined as properties of leachate may be varied depending upon the region. Still, several challenges related to leachate management and its treatments are discussed in this study. An integrated system could be a better option for treating leachate because it contains large amounts of organic and inorganic compounds. Proper leachate management would help to recover energy and value-added products (metals).

Does carbon cloth really improve thermophilic anaerobic digestion performance on a larger scale? focusing on statistical analysis and microbial community dynamics.

Currently, the phenomenon of direct interspecies electron transfer (DIET) is of great interest in the technology of anaerobic digestion (AD) due to potential performance benefits. However, the conditions for the occurrence of DIET and its limits on improving AD under conditions close to real have not been studied enough. This research is concentrated on the effect of conductive carbon cloth (R3), in comparison with a dielectric fiberglass cloth (R2) and control (R1), on the AD performance in large (90 L) thermophilic reactors, fed with a mixture of simulated organic fraction of municipal solid waste and sewage sludge. While organic loading rate (OLR) was gradually increased from 2.4 to 8.66 kg VS/(m3 day), a statistically significant (p < 0.05) difference in biogas production was observed between R1 and both R2 and R3. However, at a maximum OLR of 12.12 kg VS/(m3 day) in R3, an increase in biogas production (p < 0.05) was observed both compared to R1 (by 8.97%) and R2 (by 4.24%). The content of volatile fatty acids in R3 as a whole was the lowest, especially at the maximum OLR. Biofilm on carbon cloth was rich in syntrophic microorganisms of the genera Tepidanaerobacter, as well as Defluviitoga, capable of DIET in mixed cultures with Methanothrix, which was the most abundant methanogen in biofilm. Suspended Bifidobacterium, Fervidobacterium and Anaerobaculum were negatively affected, while Defluviitoga, Methanothermobacter and Methanosarcina, on the contrary, were positively affected by the increase in OLR and showed, respectively, a negative and positive correlation (p < 0.05) with the main AD performance parameters.

Biochemical hydrogen potential assay for predicting the patterns of the kinetics of semi-continuous dark fermentation.

The performance and kinetics response of thermophilic semi-continuous dark fermentation (DF) of simulated complex carbohydrate-rich waste was investigated at various hydraulic retention times (HRT) (2, 2.5, and 3 d) and compared with data obtained from biochemical hydrogen potential assay (BHP). A culture of Thermoanaerobacterium thermosaccharolyticum was used as the inoculum and dominated both in BHP and semi-continuous reactor. Both the modified Gompertz and first-order models described the DF kinetics well (R2 = 0.97-1.00). HRT of 2.5 d was found to be optimal in terms of maximum hydrogen production rate and hydrogen potential, which were 3.97 and 1.26 times higher, respectively, than in BHP. The hydrolysis constant was highest at HRT of 3 d and was closest to the value obtained in the BHP. Overall, BHP has been shown to be a useful tool for predicting H2 potential and the hydrolysis constant, while the maximum H2 production rate is greatly underestimated.

Comparative effect of conductive and dielectric materials on methanogenesis from highly concentrated volatile fatty acids.

Various conductive materials and their dielectric counterparts were used to get deeper insights into contribution of direct interspecies electron transfer (DIET) in improving methanogenesis from highly concentrated volatile fatty acids (12.5 g/L). Potential CH4 yield, maximum CH4 production rate and lag phase were significantly (up to 1.4, 3.9 and 2.0 times, respectively) improved with addition of stainless-steel mesh (SM) and carbon felt (CF) compared to both control and dielectric counterparts (p < 0.05). kapp increased by 82% for SM and 63% for CF compared to control (p < 0.05). Short thick pili-like structures up to 150 nm in width were formed only in CF and SM biofilms, however, were more abundant for SM. Ureibacillus and Limnochordia specific for SM biofilms, and Coprothermobacter and Ca. Caldatribacterium for CF biofilms, were considered electrogenic. Promotion of DIET by conductive materials is governed by many factors, including specificity of electrogenic groups to material surface.

Characterization of Enrichment Cultures of Anammox, Nitrifying and Denitrifying Bacteria Obtained from a Cold, Heavily Nitrogen-Polluted Aquifer.

Anammox bacteria related to Candidatus Scalindua were recently discovered in a cold (7.5 °C) aquifer near sludge repositories containing solid wastes of uranium and processed polymetallic concentrate. Groundwater has a very high level of nitrate and ammonia pollution (up to 10 and 0.5 g/L, respectively) and a very low content of organic carbon (2.5 mg/L). To assess the potential for bioremediation of polluted groundwater in situ, enrichment cultures of anammox, nitrifying, and denitrifying bacteria were obtained and analyzed. Fed-batch enrichment of anammox bacteria was not successful. Stable removal of ammonium and nitrite (up to 100%) was achieved in a continuous-flow reactor packed with a nonwoven fabric at 15 °C, and enrichment in anammox bacteria was confirmed by FISH and qPCR assays. The relatively low total N removal efficiency (up to 55%) was due to nonstoichiometric nitrate buildup. This phenomenon can be explained by a shift in the metabolism of anammox bacteria towards the production of more nitrates and less N2 at low temperatures compared to the canonical stoichiometry. In addition, the too high an estimate of specific anammox activity suggests that N cycle microbial groups other than anammox bacteria may have contributed significantly to N removal. Stable nitrite production was observed in the denitrifying enrichment culture, while no "conventional" nitrifiers were found in the corresponding enrichment cultures. Xanthomonadaceae was a common taxon for all microbial communities, indicating its exclusive role in this ecosystem. This study opens up new knowledge about the metabolic capabilities of N cycle bacteria and potential approaches for sustainable bioremediation of heavily N-polluted cold ecosystems.

Effect of Mineral Carriers on Biofilm Formation and Nitrogen Removal Activity by an Indigenous Anammox Community from Cold Groundwater Ecosystem Alone and Bioaugmented with Biomass from a "Warm" Anammox Reactor.

The complex pollution of aquifers by reduced and oxidized nitrogen compounds is currently considered one of the urgent environmental problems that require non-standard solutions. This work was a laboratory-scale trial to show the feasibility of using various mineral carriers to create a permeable in situ barrier in cold (10 °C) aquifers with extremely high nitrogen pollution and inhabited by the Candidatus Scalindua-dominated indigenous anammox community. It has been established that for the removal of ammonium and nitrite in situ due to the predominant contribution of the anammox process, quartz, kaolin clays of the Kantatsky and Kamalinsky deposits, bentonite clay of the Berezovsky deposit, and zeolite of the Kholinsky deposit can be used as components of the permeable barrier. Biofouling of natural loams from a contaminated aquifer can also occur under favorable conditions. It has been suggested that the anammox activity is determined by a number of factors, including the presence of the essential trace elements in the carrier and the surface morphology. However, one of the most important factors is competition with other microbial groups that can develop on the surface of the carrier at a faster rate. For this reason, carriers with a high specific surface area and containing the necessary microelements were overgrown with the most rapidly growing microorganisms. Bioaugmentation with a "warm" anammox community from a laboratory reactor dominated by Ca. Kuenenia improved nitrogen removal rates and biofilm formation on most of the mineral carriers, including bentonite clay of the Dinozavrovoye deposit, as well as loamy rock and zeolite-containing tripoli, in addition to carriers that perform best with the indigenous anammox community. The feasibility of coupled partial denitrification-anammox and the adaptation of a "warm" anammox community to low temperatures and hazardous components contained in polluted groundwater prior to bioaugmentation should be the scope of future research to enhance the anammox process in cold, nitrate-rich aquifers.

Brain Natriuretic Peptide (BNP) Affects Growth and Stress Tolerance of Representatives of the Human Microbiome, Micrococcus luteus C01 and Alcaligenes faecalis DOS7.

Brain natriuretic peptide (BNP) is secreted by the ventricles of the heart during overload to signal heart failure. Slight bilateral skin itching induced by BNP has been associated with response activity of the skin microbiota. In this work, we studied the effect of 25-250,000 pg BNP/mL on the growth, long-term survival, and stress (H2O2, antibiotics, salinity, heat and pH shock) resistance of human symbiont bacteria: Gram-positive Micrococcus luteus C01 and Gram-negative Alcaligenes faecalis DOS7. The effect of BNP turned out to be dose-dependent. Up to 250 pg BNP/mL made bacteria more stress resistant. At 2500 pg BNP/mL (heart failure) the thermosensitivity of the bacteria increased. Almost all considered BNP concentrations increased the resistance of bacteria to the action of tetracycline and ciprofloxacin. Both bacteria survived 1.3-1.7 times better during long-term (up to 4 months) storage. Our findings are important both for clinical medical practice and for practical application in other areas. For example, BNP can be used to obtain stress-resistant bacteria, which is important in the collection of microorganisms, as well as for the production of bacterial preparations and probiotics for cosmetology, agriculture, and waste management.

Didecyldimethylammonium Chloride- and Polyhexamethylene Guanidine-Resistant Bacteria Isolated from Fecal Sludge and Their Potential Use in Biological Products for the Detoxification of Biocide-Contaminated Wastewater Prior to Conventional Biological Treatment.

Toxic shock caused by the discharge of biocide-contaminated fecal sludge (FS) from chemical toilets to conventional wastewater treatment plants (WWTP) can be a major problem in activated sludge operation. It is necessary to develop new environmental approaches to mitigate the toxicity of biocides in order to avoid degrading the performance of WWTP. "Latrina", a chemical toilet additive containing didecyldimethylammonium chloride and polyhexamethylene guanidine, is widely used in environmentally safe toilet complexes (ESTC) on Russian railway trains to deodorize FS and control microbial activity. In this work, seven biocide-resistant bacterial strains were isolated and identified from the FS of ESTC. The values of the minimum inhibitory and bactericidal concentrations of biocides for the isolated strains were 4.5-10 times higher than for the collection microorganisms. The bacterium Alcaligenes faecalis DOS7 was found to be particularly resistant to "Latrina", the minimum inhibitory concentration of which was almost 30 times higher than recommended for ESTC. Biological products based on isolated bacterial strains proved to be effective for FS biodegradation under both aerobic and anaerobic conditions. The results of the biochemical oxygen demand test and the newly developed disk-diffusion bioassay confirmed that isolated strains contribute to reducing toxicity of biocidal agents in FS. Hyper-resistance, non-pathogenicity, and potential plant growth-promoting ability make A. faecalis DOS7 promising for use in various biological products for wastewater treatment and bioremediation of soils contaminated with biocides, as well as in agriculture to increase plant productivity.

Effects of various materials used to promote the direct interspecies electron transfer on anaerobic digestion of low-concentration swine manure.

The activation of direct interspecies electron transfer (DIET) by the supplementation of conductive materials is one of the effective and available methods to enhance anaerobic digestion (AD). Microorganisms that colonize the surface of these materials form biofilms, the study of which could provide new insights into the character of the DIET process and its effect on AD. The present study focused on AD performance, microbial community, as well as morphological and topological features of biofilms on various materials used to promote DIET during AD of low-concentration swine manure. The best AD characteristics were observed in stainless steel mesh (SM)/digested cow manure (CM) and polyester felt (PF)/digested sewage sludge (SS) combinations used as material/inoculum, respectively. Thus, potential methane yields in CM-SM and SS-PF were up to 26.4% and 26.2% higher compared to the corresponding controls. Microbial analysis of biofilms revealed the dominance of putatively syntrophic bacteria of the MBA03 group of the Limnochordia class in CM inoculated reactors, and syntrophic proteolytic bacteria of the genus Coprothermobacter and acetogenic Clostridium sensu stricto 1, known for their ability to carry out DIET, in SS inoculated reactors. Biofilms on non-conductive materials contained pili-like structures, which were observed only in SS inoculated reactors. Polyester felt tended to biofoul better than carbon felt, resulting in up to 2.8, 3.2 and 1.8 higher nucleic acid, extracellular polymeric substances, and total biomass content, respectively, depending on the inoculum. These results provide new insights into the different types of DIET that can occur in low-loaded AD systems with attached growth.

Biocides with Controlled Degradation for Environmentally Friendly and Cost-Effective Fecal Sludge Management.

Didecyldimethylammonium chloride (DDAC) and polyhexamethylene guanidine (PHMG) exhibit high antimicrobial activity and are widely used as biocidal agents in chemical toilet additives for the management of fecal sludge (FS). Disposal of such biocide-treated FS to a wastewater treatment plant (WWTP) is a major environmental problem. It is possible to reduce environmental damage through the use of biocidal agents, which easily decompose after performing their main biocidal functions. In this work, it is proposed to use the fact of a gradual increase in pH of FS from the initial 7.5 to 9.0-10.0 due to the decomposition of urea. Six biocidal compounds were selected that are capable of rapidly degrading in an alkaline environment and one that naturally degrades upon prolonged incubation. Four of them: bronopol (30 mg/L), DBNPA (500 mg/L), Sharomix (500 mg/L), and sodium percarbonate (6000 mg/L) have shown promise for environmentally friendly management of FS. In selected dosage, they successfully reduced microbial activity under both aerobic and anaerobic conditions and are cost-effective. After 10 days of incubation, degradation of the biocide occurred as measured by biological oxygen demand (BOD5) in biocide-treated FS. Such FS can be discharged to WWTP without severe damage to the activated sludge process, the need for dilution and additional procedures to neutralize toxicity.

Genome of a Novel Bacterium "Candidatus Jettenia ecosi" Reconstructed From the Metagenome of an Anammox Bioreactor.

The microbial community of a laboratory-scale bioreactor based on the anammox process was investigated by using metagenomic approaches and fluorescent in situ hybridization (FISH). The bioreactor was initially inoculated with activated sludge from the denitrifying bioreactor of a municipal wastewater treatment station. By constantly increasing the ammonium and nitrite load, a microbial community containing the novel species of anammox bacteria "Candidatus Jettenia ecosi" developed in the bioreactor after 5 years when the maximal daily nitrogen removal rate reached 8.5 g/L. Sequencing of the metagenome of anammox granules and the binning of the contigs obtained, allowed a high quality draft genome of the dominant anammox bacterium, "Candidatus Jettenia ecosi" to be assembled. Annotation of the 3.9 Mbp long genome revealed 3970 putative protein-coding genes, 45 tRNA genes, and genes for 16S/23S rRNAs. Analysis of the genome of "Candidatus Jettenia ecosi" revealed genes involved in anammox metabolism, including nitrite and ammonium transporters, copper-containing nitrite reductase, a nitrate reductase complex, hydrazine synthase, and hydrazine dehydrogenase. Autotrophic carbon fixation could be accomplished through the Wood Ljungdahl pathway. The composition of the community was investigated through a search of 16S rRNA sequences in the metagenome and FISH analysis of the anammox granules. The presence of the members of Ignavibacteriae, Betaproteobacteria, Chloroflexi and other microbial lineages reflected the complexity of the microbial processes in the studied bioreactor performed by anammox Planctomycetes, fermentative bacteria, and denitrifiers.

Calorithrix insularis gen. nov., sp. nov., a novel representative of the phylum Calditrichaeota.

A moderately thermophilic, anaerobic bacterium designated as strain KRT was isolated from a shallow-water submarine hydrothermal vent (Kunashir Island, Southern Kurils, Russia). Cells of strain KRT were thin (0.2-0.3 µm), flexible, motile, Gram-stain-negative rods of variable length. Optimal growth conditions were pH 6.6, 55 °C and 1-3 % (w/v) NaCl. Strain KRT was able to ferment a wide range of proteinaceous substrates, pyruvate, and mono-, di- and polysaccharides. The best growth occurred with proteinaceous compounds. Nitrate significantly stimulated the growth on proteinaceous substrates decreasing H2 formation, ammonium being the main product of nitrate reduction. Strain KRT did not need the presence of a reducing agent in the medium and tolerated the presence of oxygen in the gas phase up to 3 % (v/v). In the presence of nitrate, aerotolerance of isolate KRT was enhanced up to 6-8 % O2 (v/v). Strain KRT was able to grow chemolithoheterotrophically, oxidizing H2 and reducing nitrate to ammonium. Yeast extract (0.05 g l-1) was required for growth. The G+C content of the genomic DNA of strain KRT was 47.3 mol%. 16S rRNA gene sequence analysis placed isolate KRT in the phylum Calditrichaeota where it represented a novel species of a new genus, for which the name Calorithrix insularis gen. nov., sp. nov. is proposed. The type strain of Calorithrix insularis is KRT (=DSM 101605T=VKM B-3022T).