Fan Lü

Predominant Contribution of Syntrophic Acetate Oxidation to Thermophilic Methane Formation at High Acetate Concentrations

To quantify the contribution of syntrophic acetate oxidation to thermophilic anaerobic methanogenesis under the stressed condition induced by acidification, the methanogenic conversion process of 100 mmol/L acetate was monitored simultaneously by using isotopic tracing and selective inhibition techniques, supplemented with the analysis of unculturable microorganisms. Both quantitative methods demonstrated that, in the presence of aceticlastic and hydrogenotrophic methanogens, a large percentage of methane (up to 89%) was initially derived from CO(2) reduction, indicating the predominant contribution of the syntrophic acetate oxidation pathway to acetate degradation at high acid concentrations. A temporal decrease of the fraction of hydrogenotrophic methanogenesis from more than 60% to less than 40% reflected the gradual prevalence of the aceticlastic methanogenesis pathway along with the reduction of acetate. This apparent discrimination of acetate methanization pathways highlighted the importance of the syntrophic acetate-oxidizing bacteria to initialize methanogenesis from high organic loadings.

Dissolved organic matter with multi-peak fluorophores in landfill leachate.

Dissolved organic matter (DOM) sampled from municipal landfill leachate of different ages with/without anoxic or aerobic treatment, was intensively fractionated via size exclusion chromatography (SEC) and hydrophobic resins, and was studied with fluorescence excitation and emission matrix (EEM). Six fluorophores with multiple EEM peaks (fluorophore A-F) were identified based on the collected EEM spectra and validated by bi-variate analysis, principal component analysis, and parallel factor analysis, as follows (excitation wavelength Ex and emission wavelength Em): (Ex 240, 310, 360 nm, Em 460 nm), (Ex 220, 280 nm, Em 340 nm), (Ex 220, 270 nm, Em 300 nm), (Ex 220, 280 nm, Em 360 nm), (Ex 230, 320 nm, Em 420 nm) and (Ex 220, 310 nm, Em 400 nm). The spectral characteristics of these fluorophores were discussed using fractional EEM and apparent molecular weight (AMW) data obtained via SEC analysis. The triple peak flurophore A was pointed at a hydrophobic acid or hydrophobic neutral compound with a pyrenyl functional group of AMW 2500-3500 Da, which displayed an excitation wavelength at 360 nm and a fluorescence intensity ratio of 6.70(+/-1.79):1.70(+/-0.41):1 (fluorescent intensities of Ex 240:Ex 310:Ex 360 nm at Ex 460 nm). This compound is observed to be refractory in landfilling or in anoxic/aerobic treatments, and is specific to this leachate contamination. This paper revealed that the coupling of SEC and EEM can be useful to track the fluorescent DOM fraction in landfill leachate.

Metaproteomics of cellulose methanisation under thermophilic conditions reveals a surprisingly high proteolytic activity.

Cellulose is the most abundant biopolymer on Earth. Optimising energy recovery from this renewable but recalcitrant material is a key issue. The metaproteome expressed by thermophilic communities during cellulose anaerobic digestion was investigated in microcosms. By multiplying the analytical replicates (65 protein fractions analysed by MS/MS) and relying solely on public protein databases, more than 500 non-redundant protein functions were identified. The taxonomic community structure as inferred from the metaproteomic data set was in good overall agreement with 16S rRNA gene tag pyrosequencing and fluorescent in situ hybridisation analyses. Numerous functions related to cellulose and hemicellulose hydrolysis and fermentation catalysed by bacteria related to Caldicellulosiruptor spp. and Clostridium thermocellum were retrieved, indicating their key role in the cellulose-degradation process and also suggesting their complementary action. Despite the abundance of acetate as a major fermentation product, key methanogenesis enzymes from the acetoclastic pathway were not detected. In contrast, enzymes from the hydrogenotrophic pathway affiliated to Methanothermobacter were almost exclusively identified for methanogenesis, suggesting a syntrophic acetate oxidation process coupled to hydrogenotrophic methanogenesis. Isotopic analyses confirmed the high dominance of the hydrogenotrophic methanogenesis. Very surprising was the identification of an abundant proteolytic activity from Coprothermobacter proteolyticus strains, probably acting as scavenger and/or predator performing proteolysis and fermentation. Metaproteomics thus appeared as an efficient tool to unravel and characterise metabolic networks as well as ecological interactions during methanisation bioprocesses. More generally, metaproteomics provides direct functional insights at a limited cost, and its attractiveness should increase in the future as sequence databases are growing exponentially.

The use of biochar-amended composting to improve the humification and degradation of sewage sludge

Wood biochar (6%, 12% and 18% of fresh sludge weight) adding to a sludge-and-straw composting system was investigated to assess the potential of biochar as a composting amendment. Organic degradation efficiency, temporal humification profile of the water-extractable organic fraction and solid organic matter, through spectroscopic, microscopic and elementary analysis were monitored. Fluorescent excitation and emission matrix indicated that concentrations of aqueous fulvic-acid-like and humic-acid-like compounds were, respectively, 13-26% and 15-30% higher in the biochar-amended treatments, than those in the control without biochar-amended. On the first day of sludge aerobic incubation, the presence of biochar resulted in increased oxygen uptake rates of 21-37% due to its higher nano-porosity and surface area. SEM indicated that, in the biochar-amended sludge, the dense microstructure on the sludge surface disintegrated into fragments with organic fraction degraded and water lost. Results indicated that 12-18%w/w addition of wood biochar to sludge composting was recommended.

Regulating the hydrolysis of organic wastes by micro-aeration and effluent recirculation

In this study, the effects of micro-aeration and liquid recirculation on the hydrolysis of vegetable and flower wastes during two-phase solid-liquid anaerobic digestion were assessed. To accomplish this, we evaluated the hydrolysis of five batches of waste that were treated under the following conditions: anaerobic, insufficient micro-aeration (aeration for 5 min every 24 h), and sufficient micro-aeration (aeration for 5 min every 12, 4 and 1h). Hydrolysis was found to depend on the level of micro-aeration. Specifically, insufficient micro-aeration led to unstable and decreased performance. Conversely, sufficient micro-aeration promoted the hydrolysis of easily biodegradable carbohydrates and proteins, but the microbial activity was later impaired by liquid recirculation using methanogenic effluent. The hydrolysis efficiency under anaerobic conditions was comparable to the efficiency observed under sufficient micro-aeration, while the cumulative TOC of the anaerobic batch was 1.4-2.4 times higher than that of the micro-aerated batches. In addition, liquid recirculation did not have a negative effect on the development of microbial activity under anaerobic conditions, which resulted in the lignocelluloses having a higher hydrolysis efficiency.

Bacterial bioaugmentation for improving methane and hydrogen production from microalgae

BackgroundThe recalcitrant cell walls of microalgae may limit their digestibility for bioenergy production. Considering that cellulose contributes to the cell wall recalcitrance of the microalgae Chlorella vulgaris, this study investigated bioaugmentation with a cellulolytic and hydrogenogenic bacterium, Clostridium thermocellum, at different inoculum ratios as a possible method to improve CH4 and H2 production of microalgae.ResultsMethane production was found to increase by 17?~?24% with the addition of C. thermocellum, as a result of enhanced cell disruption and excess hydrogen production. Furthermore, addition of C. thermocellum enhanced the bacterial diversity and quantities, leading to higher fermentation efficiency. A two-step process of addition of C. thermocellum first and methanogenic sludge subsequently could recover both hydrogen and methane, with a 9.4% increase in bioenergy yield, when compared with the one-step process of simultaneous addition of C. thermocellum and methanogenic sludge. The fluorescence peaks of excitation-emission matrix spectra associated with chlorophyll can serve as biomarkers for algal cell degradation.ConclusionsBioaugmentation with C. thermocellum improved the degradation of C. vulgaris biomass, producing higher levels of methane and hydrogen. The two-step process, with methanogenic inoculum added after the hydrogen production reached saturation, was found to be an energy-efficiency method for hydrogen and methane production.

Multiscale visualization of the structural and characteristic changes of sewage sludge biochar oriented towards potential agronomic and environmental implication

Sewage sludge biochars were obtained at different pyrolysis temperatures from 300°C to 900°C and their macro- and microscale properties were analyzed. The biochars plant-available nutrients and humus-like substances in the water-extractable phase and fixed nutrients in the solid fraction were evaluated for their potential agronomic implications. FT-IR, Raman, XRD, XPS, and SEM techniques were used to investigate the chemical structure, functional groups, and microcrystal structure on the surface of the biochar. The results revealed minor chemical changes and dramatic mass loss in the biochar obtained at 300–500°C, whereas significant chemical changes in the biochar were obtained at 600–900°C. The concentrations of plant-available nutrients as well as fulvic- and humic-acid-like materials decreased in the biochar samples obtained at higher temperatures. These results implied that the biochar samples pyrolyzed at 300–500°C could be a direct nutrient source and used to neutralize alkaline soil. The surface area and porosity of the biochar samples increased with temperature, which increased their adsorption capacity. Rearrangement occurred at higher temperature 600–900°C, resulting in the biochar becoming increasingly polyaromatic and its graphite-like carbon becoming organized.

Comparing activated carbon of different particle sizes on enhancing methane generation in upflow anaerobic digester.

Two sizes of conductive particles, i.e. 10-20 mesh granulated activated carbon (GAC) and 80-100 mesh powdered activated carbon (PAC) were added into lab-scale upflow anaerobic sludge blanket reactors, respectively, to testify their enhancement on the syntrophic metabolism of alcohols and volatile fatty acids (VFAs) in 95days operation. When OLR increased to more than 5.8gCOD/L/d, the differences between GAC/PAC supplemented reactors and the control reactor became more significant. The introduction of activated carbon could facilitate the enrichment of methanogens and accelerate the startup of methanogenesis, as indicated by enhanced methane yield and substrate degradation. High-throughput pyrosequencing analysis showed that syntrophic bacteria and Methanosarcina sp. with versatile metabolic capability increased in the tightly absorbed fraction on the PAC surface, leading to the promoted syntrophic associations. Thus PAC prevails over than GAC for methanogenic reactor with heavy load.

Bacterial community dynamics and product distribution during pH‐adjusted fermentation of vegetable wastes

Aims:  To estimate the effect of pH on the structures of bacterial community during fermentation of vegetable wastes and to investigate the relationship between bacterial community dynamics and product distribution.

The combined effect of bacteria and Chlorella vulgaris on the treatment of municipal wastewaters

Impacts of Chlorella vulgaris with or without co-existing bacteria on the removal of nitrogen, phosphorus and organic matter from wastewaters were studied by comparing the wastewater treatment effects between an algae-bacteria consortium and a stand-alone algae system. In the algae-bacteria system, C.vulgaris played a dominant role in the removal of nitrogen and phosphorus, while bacteria removed most of the organic matter from the wastewater. When treating unsterilized wastewater, bacteria were found to inhibit the growth of algae at >231 mg/L dissolved organic carbon (DOC). Using the algae-bacteria consortium resulted in the removal of 97% NH4(+), 98% phosphorus and 26% DOC at a total nitrogen (TN) level of 29-174 mg/L. The reaction rate constant (k) values in sterilized and unsterilized wastewaters were 2.17 and 1.92 mg NH4(+)-N/(mg algal cell ·d), respectively.