Anaerobic Fermentative Bacteria Research Articles

Alkaline Pre-Fermentation Promotes Anaerobic Digestion of Enhanced Membrane Coagulation (EMC) Sludge: Performance and Microbial Community Response

Concentrating organic matter in sludge and converting it into methane through anaerobic bioconversion can improve resource recovery from domestic wastewater. Enhanced membrane coagulation (EMC) is highly efficient at concentrating organic matter, but residual coagulants (aluminum salts) can obstruct bioconversion by blocking microbial access. Limited research exists on evaluating EMC sludge bioconversion performance and addressing coagulant inhibition. This study proposes alkaline pre-fermentation to break down HO-Al-P backbones in coagulated sludge flocs, thereby improving hydrolysis and organic acid production for anaerobic digestion. Among the tested alkaline conditions (pH 9, pH 10, pH 11), pre-fermentation at pH 11 released the most organic matter (4710.0 mg/L SCOD), 20.4 times higher than without alkaline treatment. At pH 11, phosphate (61 mg/L PO43−–P) and organic acid production (2728.1 mg COD/L, with nearly 50% acetic acid) peaked, resulting in superior volatile solids removal (65.2%) and methane production (185.8 mL/g VS) during anaerobic digestion. Alkaline pre-fermentation favored alkali-tolerant bacteria such as Firmicutes and Actinobacteria, especially at pH 11, while neutrophilic Proteobacteria were suppressed. Trichococcus and Bifidobacterium, known acid producers, dominated under all conditions, with their abundance increasing at higher pH levels. Anaerobic digestion enriched fermentative bacteria like Chloroflexi and Synergistota (e.g., Thermovirga), especially in high pH reactors. Methanothrix, an acetoclastic methanogen, became the dominant methanogenic archaeon, indicating that methane production from EMC sludge primarily followed the acetoclastic methanogenesis pathway. Our findings demonstrate that alkaline pre-fermentation at pH 11 significantly enhances the hydrolysis efficiency of EMC sludge for methane recovery.

Efficient phosphate and hydrogen recovery from sludge fermentation liquid by sacrificial iron anode in electro-fermentation system

Electro-fermentation (EF) has been extensively studied for recovering hydrogen and phosphorus from waste activated sludge (WAS), while was limited for the further application due to the low hydrogen yield and phosphorus recovery efficiency. This study proposed an efficient strategy for hydrogen and vivianite recovery from the simulated sludge fermentation liquid by sacrificial iron anode in EF. The optimum hydrogen productivity and the utilization efficiency of short chain fatty acids (SCFAs) reached 45.2 mmol/g COD and 77.6% at 5 d in pH 8. Phosphate removal efficiency achieved at 90.8% at 2 d and the high crystallinity and weight percentage of vivianite (84.8%) was obtained. The functional microbes, i.e., anaerobic fermentative bacteria, electrochemical active bacteria, homo-acetogens and iron-reducing bacteria were highly enriched and the inherent interaction between the microbial consortia and environmental variables was thoroughly explored. This work may provide a theoretical basis for energy/resource recovery from WAS in the further implementation.

Effects of nutrient injection on the Xinjiang oil field microbial community studied in a long core flooding simulation device.

Microbial Enhanced Oil Recovery (MEOR) is an option for recovering oil from depleted reservoirs. Numerous field trials of MEOR have confirmed distinct microbial community structure in diverse production wells within the same block. The variance in the reservoir microbial communities, however, remains ambiguously documented. In this study, an 8 m long core microbial flooding simulation device was built on a laboratory scale to study the dynamic changes of the indigenous microbial community structure in the Qizhong Block, Xinjiang oil field. During the MEOR, there was an approximate 34% upswing in oil extraction. Based on the 16S rRNA gene high-throughput sequencing, our results indicated that nutrition was one of the factors affecting the microbial communities in oil reservoirs. After the introduction of nutrients, hydrocarbon oxidizing bacteria became active, followed by the sequential activation of facultative anaerobes and anaerobic fermenting bacteria. This was consistent with the hypothesized succession of a microbial ecological "food chain" in the reservoir, which preliminarily supported the two-step activation theory for reservoir microbes transitioning from aerobic to anaerobic states. Furthermore, metagenomic results indicated that reservoir microorganisms had potential functions of hydrocarbon degradation, gas production and surfactant production. Understanding reservoir microbial communities and improving oil recovery are both aided by this work.

Relationships between environmental factors and N-cycling microbes reveal the indirect effect of further eutrophication on denitrification and DNRA in shallow lakes

Traditional views indicate that eutrophication and subsequent algal blooms favor denitrification and dissimilatory nitrate reduction to ammonium (DNRA) in lake ecosystems. However, lakes tend to show an increasing propensity for inorganic nitrogen (N) limitation as they become more eutrophic. Thus, the influence of further eutrophication on denitrification and DNRA in eutrophic lakes are unclear due to the uncertainty of N availability. To fill this gap, we investigated the genes abundance (AOA, AOB, nirS, nirK and nrfA) and the composition of N-cycling microbes through quantitative PCR and 16S rRNA sequencing analysis, respectively, in 15 shallow eutrophic lakes of the Yangtze-Huaihe River basin, China. The results indicated that denitrification and DNRA rates could be modulated mainly by their functional gene abundances (nirS, nirK and nrfA), followed by the environmental factors (sediment total organic carbon and nitrogen). Denitrification rates significantly increased from slightly to highly eutrophic lakes, but DNRA rates were not. An explanation is that nitrification provided ample nitrate for denitrification, and this cooperative interaction was indicated by the positive correlation of their gene abundances. In addition, Pseudomonas and Anaeromyxobacter was the dominant genus mediated denitrification and DNRA, showing the potential to perform facultative anaerobic and strict anaerobic nitrate reduction, respectively. High level of dissolved oxygen might favor the facultatively aerobic denitrifiers over the obligately anaerobic fermentative DNRA bacteria in these shallow lakes. Chlorophyll a had a weak but positive effect on the gene abundances for nitrification (AOA and AOB). Further eutrophication had an indirect effect on denitrification and DNRA rates through modulating the genes abundances of N-cycling microbes.

A (S)-3-Hydroxybutyrate Dehydrogenase Belonging to the 3-Hydroxyacyl-CoA Dehydrogenase Family Facilitates Hydroxyacid Degradation in Anaerobic Bacteria.

Ketone bodies, including acetoacetate, 3-hydroxybutyrate, and acetone, are produced in the liver of animals during glucose starvation. Enzymes for the metabolism of (R)-3-hydroxybutyrate have been extensively studied, but little is known about the metabolism of its enantiomer (S)-3-hydroxybutyrate. Here, we report the characterization of a novel pathway for the degradation of (S)-3-hydroxybutyrate in anaerobic bacteria. We identify and characterize a stereospecific (S)-3-hydroxylbutyrate dehydrogenase (3SHBDH) from Desulfotomaculum ruminis, which catalyzes the reversible NAD(P)H-dependent reduction of acetoacetate to form (S)-3-hydroxybutyrate. 3SHBDH also catalyzes oxidation of d-threonine (2R, 3S) and l-allo-threonine (2S, 3S), consistent with its specificity for β-(3S)-hydroxy acids. Isothermal calorimetry experiments support a sequential mechanism involving binding of NADH prior to (S)-3-hydroxybutyrate. Homologs of 3SHBDH are present in anaerobic fermenting and sulfite-reducing bacteria, and experiments with Clostridium pasteurianum showed that 3SHBDH, acetate CoA-transferase (YdiF), and (S)-3-hydroxybutyryl-CoA dehydrogenase (Hbd) are involved together in the degradation of (S)-3-hydroxybutyrate as a carbon and energy source for growth. (S)-3-hydroxybutyrate is a human metabolic marker and a chiral precursor for chemical synthesis, suggesting potential applications of 3SHBDH in diagnostics or the chemicals industry. IMPORTANCE (R)-3-hydroxybutyrate is well studied as a component of ketone bodies produced by the liver and of bacterial polyesters. However, the biochemistry of its enantiomer (S)-3-hydroxybutyrate is poorly understood. This study describes the identification and characterization of a stereospecific (S)-3-hydroxylbutyrate dehydrogenase and its function in a metabolic pathway for the degradation of (S)-3-hydroxybutyrate as a carbon and energy source in anaerobic bacteria. (S)-3-hydroxybutyrate is a mammalian metabolic marker and a precursor for chemical synthesis and bioplastics, suggesting potential applications of these enzymes in diagnostics and biotechnology.

Natronogracilivirga saccharolytica gen. nov., sp. nov. and Cyclonatronum proteinivorum gen. nov., sp. nov., haloalkaliphilic organotrophic bacteroidetes from hypersaline soda lakes forming a new family Cyclonatronaceae fam. nov. in the order Balneolales

Two heterotrophic bacteroidetes strains were isolated as satellites from autotrophic enrichments inoculated with samples from hypersaline soda lakes in southwestern Siberia. Strain Z-1702T is an obligate anaerobic fermentative saccharolytic bacterium from an iron-reducing enrichment culture, while Ca. Cyclonatronum proteinivorum OmegaT is an obligate aerobic proteolytic microorganism from a cyanobacterial enrichment. Cells of isolated bacteria are characterized by highly variable morphology. Both strains are chloride-independent moderate salt-tolerant obligate alkaliphiles and mesophiles. Strain Z-1702T ferments glucose, maltose, fructose, mannose, sorbose, galactose, cellobiose, N-acetyl-glucosamine and alpha-glucans, including starch, glycogen, dextrin, and pullulan. Strain OmegaT is strictly proteolytic utilizing a range of proteins and peptones. The main polar lipid fatty acid in both strains is iso-C15:0, while other major components are various C16 and C17 isomers. According to pairwise sequence alignments using BLAST Gracilimonas was the nearest cultured relative to both strains (<90% of 16S rRNA gene sequence identity). Phylogenetic analysis placed strain Z-1702T and strain OmegaT as two different genera in a deep-branching clade of the new family level within the order Balneolales with genus. Based on physiological characteristics and phylogenetic position of strain Z-1702T it was proposed to represent a novel genus and species Natronogracilivirga saccharolityca gen. nov., sp. nov. (= DSMZ 109061T =JCM 32930T =VKM B 3262T). Furthermore, phylogenetic and phenotypic parameters of N. saccharolityca and C. proteinivorum gen. nov., sp. nov., strain OmegaT (=JCM 31662T, =UNIQEM U979T), make it possible to include them into a new family with a proposed designation Cyclonatronaceae fam. nov..

The gut microbiome variability of a butterflyfish increases on severely degraded Caribbean reefs

Environmental degradation has the potential to alter key mutualisms that underlie the structure and function of ecological communities. How microbial communities associated with fishes vary across populations and in relation to habitat characteristics remains largely unknown despite their fundamental roles in host nutrition and immunity. We find significant differences in the gut microbiome composition of a facultative coral-feeding butterflyfish (Chaetodon capistratus) across Caribbean reefs that differ markedly in live coral cover (∼0–30%). Fish gut microbiomes were significantly more variable at degraded reefs, a pattern driven by changes in the relative abundance of the most common taxa potentially associated with stress. We also demonstrate that fish gut microbiomes on severely degraded reefs have a lower abundance of Endozoicomonas and a higher diversity of anaerobic fermentative bacteria, which may suggest a less coral dominated diet. The observed shifts in fish gut bacterial communities across the habitat gradient extend to a small set of potentially beneficial host associated bacteria (i.e., the core microbiome) suggesting essential fish-microbiome interactions may be vulnerable to severe coral degradation.

Characteristics of microbial community in EGSB system treating with oxytetracycline production wastewater

In order to realize the efficient and stable operation of anaerobic digestion for oxytetracycline (OTC) production wastewater which contains high concentration refractory organic matters and antibiotic residues, two laboratory-scale EGSB reactors (the experimental reactor and the control reactor) were constructed for pre-treating OTC production wastewater and the complex characteristics and connections among anaerobic fermentative bacteria, methanogens and fungi were analyzed. The experimental reactor gradually increased OTC doses of 0–200 mg/L by four phases compared with the control reactor which was fed without OTC addition during 280 days’ operation. The average COD removal efficiency of 91.44% with the average OTC removal efficiency of 27.90% was achieved at OTC concentration of 200 mg/L. The addition of OTC did not affect the preponderant methanogen type, and Methanosaeta, a strict aceticlastic methanogen genus, was dominant both in working and controlling reactors on day 280. Redundancy analysis revealed that OTC and VFAs were the main environmental factors affecting the microbial communities and molecular ecological networks analysis indicated that the key genera principally belonged to Methanosaeta, Proteobacteria and Apiotrichum. Additionally, the fungi genus Apiotrichum might be related to the degradation of complex organic contaminants in OTC production wastewater treatment system.

Soehngenia longivitae sp. nov., a Fermenting Bacterium Isolated from a Petroleum Reservoir in Azerbaijan, and Emended Description of the Genus Soehngenia.

A methanogenic enrichment growing on a medium with methanol was obtained from a petroleum reservoir (Republic of Azerbaijan) and stored for 33 years without transfers to fresh medium. High-throughput sequencing of the V4 region of the 16S rRNA gene revealed members of the genera Desulfovibrio, Soehngenia, Thermovirga, Petrimonas, Methanosarcina, and Methanomethylovorans. A novel gram-positive, rod-shaped, anaerobic fermentative bacterium, strain 1933PT, was isolated from this enrichment and characterized. The strain grew at 13–55 °C (optimum 35 °C), with 0–3.0% (w/v) NaCl (optimum 0–2.0%) and in the pH range of 6.7–8.0 (optimum pH 7.0). The 16S rRNA gene sequence similarity, the average nucleotide identity (ANI) and in silico DNA–DNA hybridization (dDDH) values between strain 1933PT and the type strain of the most closely related species Soehngenia saccharolytica DSM 12858T were 98.5%, 70.5%, and 22.6%, respectively, and were below the threshold accepted for species demarcation. Genome-based phylogenomic analysis and physiological and biochemical characterization of the strain 1933PT (VKM B-3382T = KCTC 15984T) confirmed its affiliation to a novel species of the genus Soehngenia, for which the name Soehngenia longivitae sp. nov. is proposed. Genome analysis suggests that the new strain has potential in the degradation of proteinaceous components.

Long-term enriched methanogenic communities from thermokarst lake sediments show species-specific responses to warming.

Thermokarst lakes are large potential greenhouse gas (GHG) sources in a changing Arctic. In a warming world, an increase in both organic matter availability and temperature is expected to boost methanogenesis and potentially alter the microbial community that controls GHG fluxes. These community shifts are, however, challenging to detect by resolution-limited 16S rRNA gene-based approaches. Here, we applied full metagenome sequencing on long-term thermokarst lake sediment enrichments on acetate and trimethylamine at 4°C and 10°C to unravel species-specific responses to the most likely Arctic climate change scenario. Substrate amendment was used to mimic the increased organic carbon availability upon permafrost thaw. By performing de novo assembly, we reconstructed five high-quality and five medium-quality metagenome-assembled genomes (MAGs) that represented 59% of the aligned metagenome reads. Seven bacterial MAGs belonged to anaerobic fermentative bacteria. Within the Archaea, the enrichment of methanogenic Methanosaetaceae/Methanotrichaceae under acetate amendment and Methanosarcinaceae under trimethylamine (TMA) amendment was not unexpected. Surprisingly, we observed temperature-specific methanogenic (sub)species responses with TMA amendment. These highlighted distinct and potentially functional climate-induced shifts could not be revealed with 16S rRNA gene-based analyses. Unraveling these temperature- and nutrient-controlled species-level responses is essential to better comprehend the mechanisms that underlie GHG production from Arctic lakes in a warming world.

Extracellular polymeric substance decomposition linked to hydrogen recovery from waste activated sludge: Role of peracetic acid and free nitrous acid co-pretreatment in a prefermentation-bioelectrolysis cascading system

Free nitrous acid (FNA) has been recently reported to be an effective and eco-friendly inactivator for waste activated sludge (WAS), while the limited decomposition of the extracellular polymeric substance (EPS) matrix hampers resource recovery from WAS. This work employed peracetic acid (PAA) to assist FNA and explored the contribution of co-pretreatment to hydrogen recovery in a prefermentation-bioelectrolysis cascading system. The results showed that co-pretreatment led to approximately 8.8% and 20.4% increases in the exfoliation of particulate proteins and carbohydrates, respectively, from tightly bound EPS (TB-EPS) over that of sole FNA pretreatment. Electron paramagnetic resonance analysis verified that the synergistic effect of FNA, PAA and various generated free radicals was the essential process. This effect further promoted the accumulation of volatile fatty acids (VFAs) after 96 h of prefermentation, and the peak concentration in co-pretreated WAS (AD-FPWAS) was approximately 2.5-fold that in sole FNA-pretreated WAS (AD-FWAS). Subsequently, the cascading utilization of organics in the bioelectrolysis step contributed to efficient hydrogen generation. A total of 10.8 ± 0.3 mg H2/g VSS was harvested in microbial electrolysis cells (MECs) fed with AD-FPWAS, while 6.2 ± 0.1 mg H2/g VSS was obtained from AD-FWAS. X-ray photoelectron spectroscopy (XPS) revealed the effective decomposition of the phospholipid bilayer in the cytomembrane and the transformation of macromolecular organics into VFAs and hydrogen in the cascading system. Further microbial community analysis demonstrated that co-pretreatment enhanced the accumulation of functional consortia, including anaerobic fermentative bacteria (AFB, 28.1%), e.g., Macellibacteroides (6.3%) and Sedimentibacter (6.9%), and electrochemically active bacteria (EAB, 57.0%), e.g., Geobacter (39.0%) and Pseudomonas (13.6%), in the prefermentation and MEC steps, respectively. The possible synergetic and competitive relationships among AFB, EAB, homo-acetogens, nitrate-reducing bacteria and methanogens were explored by molecular ecological network analysis. From an environmental and economic perspective, this promising FNA and PAA co-pretreatment approach provides new insight for energy recovery from WAS biorefineries.

A gene cluster for taurine sulfur assimilation in an anaerobic human gut bacterium.

Aminoethylsulfonate (taurine) is widespread in the environment and highly abundant in the human body. Taurine and other aliphatic sulfonates serve as sulfur sources for diverse aerobic bacteria, which carry out cleavage of the inert sulfonate C-S bond through various O2-dependent mechanisms. Taurine also serves as a sulfur source for certain strict anaerobic fermenting bacteria. However, the mechanism of C-S cleavage by these bacteria has long been a mystery. Here we report the biochemical characterization of an anaerobic pathway for taurine sulfur assimilation in a strain of Clostridium butyricum from the human gut. In this pathway, taurine is first converted to hydroxyethylsulfonate (isethionate), followed by C-S cleavage by the O2-sensitive isethionate sulfo-lyase IseG, recently identified in sulfate- and sulfite-reducing bacteria. Homologs of the enzymes described in this study have a sporadic distribution in diverse strict and facultative anaerobic bacteria, from both the environment and the taurine-rich human gut, and may enable sulfonate sulfur acquisition in certain nutrient limiting conditions.

Evaluating indigenous diversity and its potential for microbial methane generation from thermogenic coal bed methane reservoir

Microbial enhanced coal bed methane process has emerged as cheaper, feasible and environment friendly approach for enhancing coal bed methane (CBM) generation worldwide.Despite the fact that India has tremendous potential for CBM, there are many challenges towards microbial translation of CBM. Therefore, to explore the possibility of microbial enhanced CBM generation from the deep thermogenic CBM reservoir, diversity of the microbial community was investigated using the 16S rRNA gene amplification methods. Additionally, a culture dependent analysis from the formation water of the reservoir was performed to access the ability of the indigenous microbes to produce methane from coal when enriched with suitable nutrient supplements. Culture independent data revealed the nature of communities i.e. bacterial and archaeal domain, majorly from thermophilic profile. The bacterial community was dominated by proteobacterial and Firmicutes taxa, while archaeal community was dominated by Euryarchaeota taxa (acetoclastic methanogens). The enrichment cultures derived from the formation water were able to produce methane (1002 µmol/g coal) at 45 °C. The 16S rRNA gene analysis depicted that the enriched microbial culture was dominated with anaerobic thermophilic fermentative bacteria of Clostridiales family and acetoclastic Methanosarcinales methanogens. The present study depicts the biogenic methane generation in the thermogenic-gas containing CBM reservoir could be enhanced by stimulating the indigenous microorganisms through providing suitable nutrient supplements. Present investigation is likewise the prime response to deliberate about the capability of thermogenic CBM reservoir (temperature > 40 °C) in India for biogenic methane generation through culture independent and culture dependent examination.

Draft Genome Sequence of a Fermenting Bacterium, "Sphaerochaeta halotolerans" 4-11T, from a Low-Temperature Petroleum Reservoir in Russia.

The draft genome sequence of an anaerobic fermenting bacterium, "Sphaerochaeta halotolerans" strain 4-11T, isolated from formation water of a low-temperature petroleum reservoir in Russia is presented. The genome is annotated to elucidate the taxonomic position of the strain 4-11T and to extend the public genome database.

Progress toward full scale application of the anaerobic side-stream reactor (ASSR) process

In order to reduce the investment costs of the anaerobic side-stream reactor (ASSR) process coupled with an activated sludge system and promote the full scale application, the impact of 1 d anaerobic solid retention time (SRTASSR) and 100% interchange ratio (IR) has been investigated on sludge reduction, carbon and nutrient removal efficiency and microbial community, in a sequencing batch reactor (SBR)-ASSR system. The SBR-ASSR achieved good removal efficiencies in COD (91.5 ± 3.4%), ammonium nitrogen (98.8 ± 0.5%), total nitrogen (87.9 ± 4.9%) and phosphate (92.8 ± 6.7). The sludge yield of the system was 0.1648 g TSS g−1COD; 54% lower compared to a conventional activated sludge (CAS) system. Real time quantitative polymerase chain reaction (q-PCR) showed an abundance of hydrolyzing and fermentative bacteria. Comparison at class and genus level confirmed an abundance of anaerobic hydrolyzing and fermentative bacteria, denitrifying bacteria able to simultaneous perform nitrogen and phosphate removal and phosphate accumulating organisms.

Pleomorphochaeta naphthae sp. nov., a new anaerobic fermentative bacterium isolated from an oil field.

A novel anaerobic fermentative bacterium, strain SEBR 4209T, was isolated from a water sample of a Congolese oil field. Strain SEBR 4209T is phylogenetically related to the genus Pleomorphochaeta, in the family Spirochaetaceae. Its closest relatives are Pleomorphochaeta caudata SEBR 4223T (94.5 % 16S rRNA gene sequence similarity) and Pleomorphochaeta multiformis MO-SPC2T (94.3 % similarity). Like the other members of this genus, cells have a pleomorphic morphology, in particular an annular shape and long stalks. Optimal growth was observed at 37 °C, at pH between 6.8 and 7.0, and with 40 g l-1 NaCl. This strain was only able to grow by fermentation of carbohydrates. The fermentation products from glucose utilization were acetate, ethanol, CO2 and H2. Predominant fatty acids were C14 : 0, C14 : 0 DMA, C16 : 0 and C16 : 1ω7c. The major polar lipids were phosphoglycolipids, phospholipids and glycolipids. The G+C content of the DNA was 29.6 mol%. Based on phenotypic characteristics and phylogenetic traits, strain SEBR 4209T is considered to represent a novel species of the genus Pleomorphochaeta, for which the name Pleomorphochaetanaphthae sp. nov. is proposed. The type strain is SEBR 4209T (=DSM 104684T=JCM 31871T).

Sludge reduction and microbial structures of aerobic, micro-aerobic and anaerobic side-stream reactor coupled membrane bioreactors

An anoxic/oxic membrane bioreactor (MBR) and three side-stream reactor (SSR) coupled membrane bioreactors were operated in parallel to investigate effects of dissolved oxygen (DO) level in SSR on sludge reduction and microbial community structure of SSR-MBRs. The four MBRs were equally efficient in COD and ammonium nitrogen removal. The anaerobic and micro-aerobic SSR favored nitrogen removal through denitrification, simultaneous nitrification and denitrification and autochthonous substrate release as carbon source. The micro-aerobic SSR achieved greatly higher sludge reduction efficiency (61.1%) than anaerobic (37.3%) and aerobic SSR (7.9%). Micro-aerobic SSR obtained the highest endogenous decay constant (0.035 d−1) compared to anaerobic (0.023 d−1) and aerobic SSR (0.015 d−1). High-throughput sequencing results revealed that anaerobic SSR enriched hydrolytic and fermentative bacteria, aerobic environment favored the growth of slow-growing bacteria, and micro-aerobic SSR stimulated biological activities of both anaerobic and aerobic bacteria.

Effect of salinity on diazotrophic activity and microbial composition of phototrophic communities from Bitter-1 soda lake (Kulunda Steppe, Russia).

Bitter-1 is a shallow hypersaline soda lake in Kulunda Steppe (Altai region, Russia). During a study period between 2005 and 2016, the salinity in the littoral area of the lake fluctuated within therange from 85 to 400g/L (in July of each year). Light-dependent nitrogen fixation occurred in this lake up to the salt-saturating conditions. The rates increased with a decrease in salinity, both under environmental conditions and in laboratory simulations. The salinities below 100g/L were favorable for light-dependent nitrogen fixation, while the process was dramatically inhibited above 200g/L salts. The analysis of nifH genes in environmental samples and in enrichment cultures of diazotrophic phototrophs suggested that anaerobic fermenting and sulfate-reducing bacteria could participate in the dark nitrogen fixation process up to soda-saturating conditions. However, we cannot exclude the possibility that haloalkaliphilic nonheterocystous cyanobacteria (Euhalothece sp. and Geitlerinema sp.) and anoxygenic purple sulfur bacteria (Ectothiorhodospira sp.) might also play a role in the process at light conditions. The heterocystous cyanobacterium Nodularia sp. develops at low salinity (below 80g/L) that is not characteristic for Bitter-1 Lake and thus does not make a significant contribution to the nitrogen fixation in this lake.

Combined bioaugmentation with anaerobic ruminal fungi and fermentative bacteria to enhance biogas production from wheat straw and mushroom spent straw

Bioaugmentation with anaerobic ruminal fungi and a pool of hydrogen-producing fermenting bacteria was tested on wheat straw (WS) and mushroom spent straw (MSS) with the aim of improving anaerobic digestion performance. Batch tests were set up to simulate a Bioaugmentation Anaerobic Digestion (BAD) treatment comparing single- (I-BAD) and two-stage (II-BAD) process configurations, at two reactor scales, 120 and 1200 ml (×10). In both cases, higher CH4 cumulative production was obtained in the II-BAD configuration on WS (65.1 ± 8.9 Nml and 922 ± 73.8 Nml respectively). The II-BADx10 tests allowed increasing CH4 production (≃290% and ≃330% on WS and MSS, respectively) when compared to the unaugmented condition. Final results highlighted the achievable advantages of the two stage configuration in terms of CH4 production enhancement. Microbial community investigations confirmed the efficiency of the bioaugmentation treatment and revealed that such a result was mainly related to the Methanosarcinales increase, mostly composed by Methanosaeta.

Enhanced sulfide removal and bioelectricity generation in microbial fuel cells with anodes modified by vertically oriented nanosheets

ABSTRACTAnode materials and structures are of critical importance for microbial fuel cells (MFCs) recovering energy from toxic substrates. Carbon-fiber-felt anodes modified by layers of vertically oriented TiO2 and Fe2O3 nanosheets were applied in the present study. Enhanced sulfide removal efficiencies (both over 90%) were obtained after a 48-h operation, with maximum power densities improved by 1.53 and 1.36 folds compared with MFCs with raw carbon-fiber-felt anode. The modified anodes provided more active sites for microbial adhesion with increasing biomass densities. High-throughput 16S rRNA gene sequencing analysis also indicated the increase in microbial diversities. Bacteroidetes responsible for bioelectricity generation with Thiobacillus and Spirochaeta dominating sulfide removal were found in the MFCs with the modified anodes, with less anaerobic fermentative bacteria as Firmicutes appeared. This indicates that the proposed materials are competitive for applications of MFCs generating bioelectricity from toxic sulfide.