Electrical stress and acid orange 7 synergistically clear the blockage of electron flow in the methanogenesis of low-strength wastewater.

Energy recovery from low-strength wastewater through anaerobic methanogenesis is constrained by limited substrate availability. The development of efficient methanogenic communities is critical but challenging. Here we develop a strategy to acclimate methanogenic communities using conductive carrier (CC), electrical stress (ES), and Acid Orange 7 (AO7) in a modified biofilter. The synergistic integration of CC, ES, and AO7 precipitated a remarkable 72-fold surge in methane production rate compared to the baseline. This increase was attributed to an altered methanogenic community function, independent of the continuous presence of AO7 and ES. AO7 acted as an external electron acceptor, accelerating acetogenesis from fermentation intermediates, restructuring the bacterial community, and enriching electroactive bacteria (EAB). Meanwhile, CC and ES orchestrated the assembly of the archaeal community and promoted electrotrophic methanogens, enhancing acetotrophic methanogenesis electron flow via a mechanism distinct from direct electrochemical interactions. The collective application of CC, ES, and AO7 effectively mitigated electron flow impediments in low-strength wastewater methanogenesis, achieving an additional 34% electron recovery from the substrate. This study proposes a new method of amending anaerobic digestion systems with conductive materials to advance wastewater treatment, sustainability, and energy self-sufficiency.

Chronic effects of benzalkonium chlorides on short chain fatty acids and methane production in semi-continuous anaerobic digestion of waste activated sludge.

As an emerging pollutant, benzalkonium chlorides (BACs) potentially enriched in waste activated sludge (WAS). However, the microbial response mechanism under chronic effects of BACs on acidogenesis and methanogenesis in anaerobic digestion (AD) has not been clearly disclosed. This study investigated the AD (by-)products and microbial evolution under low to high BACs concentrations from bioreactor startup to steady running. It was found that BACs can lead to an increase of WAS hydrolysis and fermentation, but a disturbance to acidogenic bacteria also occurred at low BACs concentration. A noticeable inhibition to methanogenesis occurred when BAC concentration was up to 15 mg/g TSS. Metagenomic analysis revealed the key genes involved in acetic acid (HAc) biosynthesis (i.e. phosphate acetyltransferase, PTA), ß-oxidation pathway (acetyl-CoA C-acetyltransferase) and propionic acid (HPr) conversion was slightly promoted compared with control. Furthermore, BACs inhibited the acetotrophic methanogenesis (i.e. acetyl-CoA synthetase), especially BAC concentration was up to 15 mg/g TSS, thereby enhanced short chain fatty acids (SCFAs) accumulation. Overall, chronic stimulation of functional microorganisms with increasing concentrations of BACs impact WAS fermentation.

Efficient methane production from waste activated sludge and Fenton-like pretreated rice straw in an integrated bio-electrochemical system.

Integrated microbial electrolysis cell-anaerobic digestion (MEC-AD) systems have demonstrated potential advantages for methane production in the presence of small amounts of residual inhibitors. In this study, a series of tests were conducted to analyse the acidification and methanogenesis performance of pretreated rice straw (RS) in anaerobic digestion (AD) and MEC-AD systems after the addition of Fenton-like reagents. The results indicated that the short-chain acids (SCFAs) accumulations reached 2284.64 ± 21.57 mg COD/L with a dosage ratio of 1/4 (g RS/g VSS sludge) in the MEC-AD system and that methane production increased by 63.8% compared with that of an individual AD system. In the interim, the net energy output reached 1.09 × 103 J/g TCOD, which was 1.23 times higher than that of the AD system. The residual Fe3+/Fe2+ in the pretreatment reagent was capable of promoting acidification and methanogenesis in sludge and RS fermentation. The RS hydrolysis products could constrain methanogenesis, which can be mitigated by introducing an MEC. The microbiological analyses revealed that the MEC strongly increased the enrichment of hydrogenotrophic methanogens, especially Methanobacterium (61.16%). Meanwhile, the Syntrophomonas and Acetobacterium abundances increased to 2.81% and 2.65%, respectively, which suggested the reinforcement of acetogenesis and methanogenesis. Therefore, the enhanced hydrogenotrophic methanogens might have served as the key for enhancing the efficiency of methanogenesis due to the introduction of an MEC.

Alkaline aided thermophiles pretreatment of waste activated sludge to increase short chain fatty acids production: Microbial community evolution by alkaline on hydrolysis and fermentation.

Adding alkaline into an anaerobic waste activated sludge (WAS) fermentation with thermophilic bacteria pretreatment could efficiently improve short-chain fatty acids (SCFAs) accumulation to 3550 ± 120 mg COD/L. The acidification rate in combined test was 21.2%, while that was 15.6% and 10.7% in sole thermophilic bacteria pretreatment and control tests respectively. Four distinct groups of microbes could be identified with noticeable shifts using the combined pretreatments, and tremendous effects were analyzed on organic content especially of the soluble proteins and SCFAs concentrations. Particularly, alkaline addition would significantly change the functional microbial structures, including the decrease of Caloramator with the function of thermophilic proteolytic and the increase of Acidobacteria TM7 and Petrimonas sp. The results above suggested that alkaline addition could decrease the hydrolytic substances consume by thermotolerance bacteria and final improve SCFAs accumulation in fermentation process.

Enhanced short-chain fatty acids production from waste activated sludge with alkaline followed by potassium ferrate treatment.

This study reported an efficient approach, i.e., alkaline followed by potassium ferrate (PF) pretreatment, to enhance short chain fatty acids (SCFAs) production from waste activated sludge anaerobic fermentation process. The optimum condition was initial pH of 10.0 and PF dosage of 28 mg Fe(VI)/g total suspended solid, with the highest SCFAs production of 382 mg chemical oxygen demand/g volatile suspended solid, which was 2.03 and 2.06 times higher than that of corresponding sole treatments. It was found that the alkaline + PF treatment could provide more soluble substrates for subsequent acidification process by accelerating disruption of both microbial cells and extracellular polymeric substances. And the alkaline + PF treatment also benefited to the activity promotion of specific hydrolases and inhibition of methanogens. Besides, the abundances of microorganisms related to SCFAs production, such as Proteiniclasticum and Macellibacteroides, were increased greatly, whereas the main SCFAs consumer, Proteobacteria, was decreased from 29.1% to 14.4%.

Potassium ferrate addition as an alternative pre-treatment to enhance short-chain fatty acids production from waste activated sludge.

A potentially practical technology based on ferrate (VI), i.e. potassium ferrate (PF), pretreatment integrated into waste activated sludge (WAS) anaerobic fermentation has been presented to greatly enhance short-chain fatty acids (SCFAs) production with a shortened fermentation time. The maximum production of SCFAs, 343mg chemical oxygen demand/g volatile suspended solid with acetic acid proportion of 48.2%, was obtained with PF dosage of 56mg Fe(VI)/g total suspended solid within 5days, which was increased to 5.72times compared to that of control. The mechanism study showed that PF accelerated the release rate of both intracellular and extracellular constituents. And the activities of key hydrolytic enzymes were much improved with PF addition. Moreover, PF positively enriched the abundance of microorganisms responsible for WAS hydrolysis and SCFAs production, especially acetic acid-forming characteristic genera such as Petrimonas, Fusibacter and Acetoanaerobium. Besides, the incubation time of acidogenesis and methanogenesis were separated by PF.

Clarification of phosphorus fractions and phosphorus release enhancement mechanism related to pH during waste activated sludge treatment.

This study aimed to clarify phosphorus (P) fractions in waste activated sludge (WAS) and explore release performance and enhancement mechanism of different P species related to pH. Results showed that inorganic P (IP) was the major P fraction in raw sludge (87.86% of total solid P), and non-apatite inorganic P (NAIP), the most labile P forms, occupied 81.30% of IP, suggesting that WAS could be selected as potential substitution of phosphate rock. The optimized acid and alkaline conditions were pH=4 and pH=12 for molybdate reactive P accumulation, increased by 311.20mg/L and 479.18mg/L compared to raw sludge, which were 3.80 and 5.84 times higher than that of control, respectively. The mechanism study demonstrated that high pH promoted NAIP release, and apatite P was sensitive to low pH. Moreover, the releasable and recoverable P depended on both fractions of different P species in sludge and pH adjustment for sludge treatment.

Lignocellulosic saccharification by a newly isolated bacterium, Ruminiclostridium thermocellum M3 and cellular cellulase activities for high ratio of glucose to cellobiose.

BACKGROUND: Lignocellulosic biomass is one of earth's most abundant resources, and it has great potential for biofuel production because it is renewable and has carbon-neutral characteristics. Lignocellulose is mainly composed of carbohydrate polymers (cellulose and hemicellulose), which contain approximately 75 % fermentable sugars for biofuel fermentation. However, saccharification by cellulases is always the main bottleneck for commercialization. Compared with the enzyme systems of fungi, bacteria have evolved distinct systems to directly degrade lignocellulose. However, most reported bacterial saccharification is not efficient enough without help from additional ß-glucosidases. Thus, to enhance the economic feasibility of using lignocellulosic biomass for biofuel production, it will be extremely important to develop a novel bacterial saccharification system that does not require the addition of ß-glucosidases. RESULTS: In this study, a new thermophilic bacterium named Ruminiclostridium thermocellum M3, which could directly saccharify lignocellulosic biomass, was isolated from horse manure. The results showed that R. thermocellum M3 can grow at 60 °C on a variety of carbon polymers, including microcrystalline cellulose, filter paper, and xylan. Upon utilization of these substrates, R. thermocellum M3 achieved an oligosaccharide yield of 481.5 ± 16.0 mg/g Avicel, and a cellular ß-glucosidase activity of up to 0.38 U/mL, which is accompanied by a high proportion (approximately 97 %) of glucose during the saccharification. R. thermocellum M3 also showed potential in degrading natural lignocellulosic biomass, without additional pretreatment, to oligosaccharides, and the oligosaccharide yields using poplar sawdust, corn cobs, rice straw, and cornstalks were 52.7 ± 2.77, 77.8 ± 5.9, 89.4 ± 9.3, and 107.8 ± 5.88 mg/g, respectively. CONCLUSIONS: The newly isolated strain R. thermocellum M3 degraded lignocellulose and accumulated oligosaccharides. R. thermocellum M3 saccharified lignocellulosic feedstock without the need to add ß-glucosidases or control the pH, and the high proportion of glucose production distinguishes it from all other known monocultures of cellulolytic bacteria. R. thermocellum M3 is a potential candidate for lignocellulose saccharification, and it is a valuable choice for the refinement of bioproducts.

Role of extracellular polymeric substances in enhancement of phosphorus release from waste activated sludge by rhamnolipid addition.

This study investigated the role of extracellular polymeric substances (EPSs) in enhanced performance of phosphorus (P) release from waste activated sludge (WAS) by adding rhamnolipid (RL). Results showed that compared to WAS without pretreatment, the released PO4(3-)-P increased with RL addition from 0 to 0.2 g/gTSS (total suspended solid), and increased by 208% under the optimal condition (0.1 g RL/g TSS and 72-h fermentation time). The cumulative PO4(3-)-P was better fitted with pseudo-first-order kinetic model. Moreover, the contents of metal ions increased in liquid but decreased in EPSs linearly with RL addition increasing, and WAS solubilizations were positively correlated with the released metal ions. The enhanced total dissolved P mainly came from cells and others (69.39%, 2.27-fold higher than that from EPSs), and PO4(3-)-P was the main species in both liquid and loosely bound EPSs, but organic P should be non-negligible in tightly bound EPSs.