Regulating microbial redox reactions towards enhanced removal of refractory organic nitrogen from wastewater.

Biotechnology for wastewater treatment is mainstream and effective depending upon microbial redox reactions to eliminate diverse contaminants and ensure aquatic ecological health. However, refractory organic nitrogen compounds (RONCs, e.g., nitro-, azo-, amide-, and N-heterocyclic compounds) with complex structures and high toxicity inhibit microbial metabolic activity and limit the transformation of organic nitrogen to inorganic nitrogen. This will eventually result in non-compliance with nitrogen discharge standards. Numerous efforts suggested that applying exogenous electron donors or acceptors, such as solid electrodes (electrostimulation) and limited oxygen (micro-aeration), could potentially regulate microbial redox reactions and catabolic pathways, and facilitate the biotransformation of RONCs. This review provides comprehensive insights into the microbial regulation mechanisms and applications of electrostimulation and micro-aeration strategies to accelerate the biotransformation of RONCs to organic amine (amination) and inorganic ammonia (ammonification), respectively. Furthermore, a promising approach involving in-situ hybrid anaerobic biological units, coupled with electrostimulation and micro-aeration, is proposed towards engineering applications. Finally, employing cutting-edge methods including multi-omics analysis, data science driven machine learning, technology-economic analysis, and life-cycle assessment would contribute to optimizing the process design and engineering implementation. This review offers a fundamental understanding and inspiration for novel research in the enhanced biotechnology towards RONCs elimination.

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.

Machine learning framework for intelligent aeration control in wastewater treatment plants: Automatic feature engineering based on variation sliding layer.

Intelligent control of wastewater treatment plants (WWTPs) has the potential to reduce energy consumption and greenhouse gas emissions significantly. Machine learning (ML) provides a promising solution to handle the increasing amount and complexity of generated data. However, relationships between the features of wastewater datasets are generally inconspicuous, which hinders the application of artificial intelligence (AI) in WWTPs intelligent control. In this study, we develop an automatic framework of feature engineering based on variation sliding layer (VSL) to control the air demand precisely. Results demonstrated that using VSL in classic machine learning, deep learning, and ensemble learning could significantly improve the efficiency of aeration intelligent control in WWTPs. Bayesian regression and ensemble learning achieved the highest accuracy for predicting air demand. The developed models with VSL-ML models were also successfully implemented under the full-scale wastewater treatment plant, showing a 16.12 % reduction in demand compared to conventional aeration control of preset dissolved oxygen (DO) and feedback to the blower. The VSL-ML models showed great potential to be applied for the precision air demand prediction and control. The package as a tripartite library of Python is called wwtpai, which is freely accessible on GitHub and CSDN to remove technical barriers to the application of AI technology in WWTPs.

Synergistic between autotrophic and heterotrophic microorganisms for denitrification using bio-S as electron donor.

In recent years, biological sulfur (bio-S) was employed in sulfur autotrophic denitrification (SAD) in which autotrophic Thiobacillus denitrificans and heterotrophic Stenotrophomonas maltophilia played a key role. The growth pattern of T.denitrificans and S.maltophilia exhibited a linear relationship between OD600 and CFU when OD600 < 0.06 and <0.1, respectively. When S.maltophilia has applied alone, the NorBC and NosZ were undetected, and denitrification was incomplete. The DsrA of S.maltophilia could produce sulfide as an alternative electron donor for T.denitrificans. Even though T.denitrificans had complete denitrification genes, its efficiency was low when used alone. The interaction of T.denitrificans and S.maltophilia reduced nitrite accumulation, leading to complete denitrification. A sufficient quantity of S.maltophilia may trigger the autotrophic denitrification activity of T.denitrificans. When the colony-forming units (CFU) ratio of S.maltophilia to T.denitrificans was reached at 2:1, the highest denitrification performance was achieved at 2.56 and 12.59 times higher than applied alone. This research provides a good understanding of the optimal microbial matching for the future application of bio-S.

Hybrid bioreactor built-in with fixed bio-carriers for denitrification with low C/N ratio: Hydrodynamic optimization and microbial divergence.

Hydrodynamics played an important role in the design and operation of bioreactors for wastewater treatment. In this work, an up-flow anaerobic hybrid bioreactor built-in with fixed bio-carriers was designed and optimized using computational fluid dynamics (CFD) simulation. The results indicated that the flow regime involving with vortex and dead zone was greatly affected by the positions of water inlet and bio-carrier modules. The ideal hydraulic features were obtained when the water inlet and bio-carrier modules located 9 cm and 60 cm above the bottom of reactor. Using the optimum hybrid system for nitrogen removal from wastewater with low carbon-to-nitrogen ratio (C/N = 3), the denitrification efficiency could reach 80.9 ± 0.4%. Illumina sequencing of 16S rRNA gene amplicons revealed that the microbial community divergence occurred among the biofilm on bio-carrier, the suspended sludge phase and the inoculum. Especially, the relative abundance of denitrifying genera Denitratisoma in the biofilm of bio-carrier reaches 5.73%, 6.2 times higher than that in the suspended sludge, implying the imbedded bio-carrier was conductive to enrich the specific denitrifiers to polish the denitrification performance with low carbon source. This work provided an effective method for the design optimization of bioreactor based on CFD simulation, and developed a hybrid reactor with fixed bio-carrier for nitrogen removal from wastewater with low C/N ratio.

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.

Enhancement of methane production from waste activated sludge using hybrid microbial electrolysis cells-anaerobic digestion (MEC-AD) process - A review.

Hybrid microbial electrolysis cells-anaerobic digestion (MEC-AD) was proved to increase methane productivity and methane yield of waste activated sludge (WAS) by establishing direct interspecies electron transfer method and enriching functional microorganisms. This review first summarized the pretreatment methods of WAS for MEC-AD and then reviewed the reactor configurations, operation parameters, and the economic benefit of MEC-AD. Furthermore, the enhancement mechanisms of MEC-AD were reviewed based on the analysis of thermodynamics and microbial community. It was found that the decrease of hydrogen partial pressure due to the hydrogenotrophic methanogens enriched in cathodic biofilm and direct interspecies electron transfer between exoelectrogens and anode were the core mechanisms for improving acidogenesis, acetogenesis, and methanogenesis. Finally, the potentially technological issues that need to be addressed to increase energy efficiency in large-scale MEC-AD processes were discussed.

Model-based mid-infrared spectroscopy for on-line monitoring of volatile fatty acids in the anaerobic digester.

The performance of anaerobic digestion is significantly governed by the concentration of volatile fatty acids (VFAs). Though the titration and near-infrared spectroscopy have been used to measure the VFAs in the digester, there is still lack of the establishment of on-line monitoring of VFAs in practical application. An effective quantification method based on mid-infrared (MIR) spectroscopy was developed, and used to measure the concentrations of VFAs in the anaerobic bioreactor nondestructively in parallel. The wavelet denoising (WD) spectra were used as the spectral preprocessing option. Compared with other pretreatment methods, the established calibration model built by WD spectra showed satisfactory results. Further, the model was verified using high performance liquid chromatography (HPLC), and predictions were made using real reactor effluent samples. Based on this theoretical work, a set of equipment for the in-situ online monitoring of VFAs was designed, which has high feasibility and effectively solves the problems with the current VFAs online monitoring process. These results provide a new solution for on-line monitoring of the anaerobic digestion, and have great potential for practical application.

Stepwise freezing-thawing treatment promotes short-chain fatty acids production from waste activated sludge.

Waste activated sludge (WAS), as the byproducts of wastewater treatment plants, has been greatly produced. With high cost and environmental risk of WAS disposal, to explore a low-cost and environment-friendly technology has been a great challenge. Considering that WAS is a collection of organic matters, anaerobic fermentation has been selected as a sustainable way to simultaneously recover resources and reduce environmental pollution. To recover short-chain fatty acids (SCFAs) has gained great concern because of the high value-added application and high-efficiency production process. Considering the temperature in some areas of the world can reach to below 0 °C, this study proposed an efficient strategy, i.e., stepwise freezing and thawing treatment, to promote SCFAs production. The maximal production of SCFAs, i.e., 246 mg COD/g volatile suspended solid, was obtained with the shortened retention time of five days. Mechanistic studies showed that the solubilization of both extracellular polymeric substances (EPSs) and microbial cells could be accelerated, with the EPSs removal of 58.3% for proteins and 59.0% for polysaccharides. Also, the hydrolysis process was promoted to provide more substrates for subsequent acidogenisis, and the functional microorganisms, such as Romboutsia, Paraclostridium, Macellibacteroides and Conexibacter, were greatly enriched, with a total abundance of 26.2%. Moreover, compared to control, methanogenesis was inhibited at a shortened sludge retention time (e.g., five days), which benefited to the accumulation of SCFAs, but the methane production was increased by 25.2% at a longer sludge retention time (e.g., ten days). Thus, these findings of this work may provide some new solutions for the enhanced resource recovery from WAS, and further for carbon-neutral operation of wastewater treatment plants.

Recent advancements in azo dye decolorization in bio-electrochemical systems (BESs): Insights into decolorization mechanism and practical application.

Recent advances in bio-electrochemical systems (BESs) for azo dye removal are gaining momentum due to having electrode biocarrier and electro-active bacteria that could stimulate decolorization via extracellular electron transfer. Enhanced decolorization performance is observed in most laboratory studies, indicating the great potential of BESs as an alternative to the traditional biological processes or serving as a pre-/post-processing unit to improve the performance of biological processes. It is proven more competitive in environmental friendly than physicochemical methods. While, the successful application of BESs to azo dye-containing wastewater remediation requires a deeper evaluation of its performance, mechanism and typical attributes, and a comprehensive potential evaluation of BESs practical application in terms of economic analysis and technical optimizations. This review is organized to address BESs as a practical option for azo dye removal by analyzing the decolorization mechanisms and involved functional microorganisms, followed by the comparisons of device configurations, operational conditions, and economic evaluation. It further highlights the current hurdles and prospects for the abatement of azo dyes via BES related techniques.

Stepwise alkaline treatment coupled with ammonia stripping to enhance short-chain fatty acids production from waste activated sludge.

To recover resource from waste activated sludge (WAS) is of great significance. This study proposed a promising way, i.e., stepwise alkaline treatment coupled with ammonia stripping, to remarkably enhance short-chain fatty acids (SCFAs) production from WAS anaerobic digestion. The maximal production of SCFAs, with the value of 323 mg COD/g volatile suspended solid, was obtained with first initial pH = 10 adjustment followed by second initial pH = 10 adjustment on third day coupled with ammonia stripping. Mechanistic studies showed that solubilization of both extracellular polymeric substances and cells could be accelerated by stepwise initial pH = 10 adjustment. However, without ammonia stripping, the activities of either acidogens or methanogens could be inhibited by free ammonia formed under alkaline conditions; positively, anaerobes related to SCFAs production were enriched with ammonia stripping. Moreover, the proposed strategy can simultaneously achieve nitrogen and carbon recovery, providing some solutions for the carbon-neutral operation of wastewater treatment plants.

Responses of anaerobic digestion of waste activated sludge to long-term stress of benzalkonium chlorides: Insights to extracellular polymeric substances and microbial communities.

Quaternary ammonium compounds have gained widespread attention due to their extensive enrichment in waste activated sludge (WAS) and potentially adverse effect to anaerobes. This study selected benzalkonium chlorides (BACs) as model to reveal the responses of anaerobic digestion of WAS to long-term stress of BACs. Results showed that the solubilization enhancement of WAS contributed by BACs was the acceleration of cell lysis, rather than the disruption of extracellular polymeric substances, and the accumulation improvement of short chain fatty acids (SCFAs) attributed to hydrolysis improvement and methanogenesis inhibition at either medium -or high level of BACs. In addition, a low level had no significant effect on the production of methane compared to control, with averages of 0.059 and 0.055 m3/(m3·d), respectively, whereas a medium level reduced methane production to 20% of control, and a high level almost completely inhibited methanogenesis. Correspondingly, BACs could shift microbial communities related to SCFAs and methane productions. For the bacterial community, a high level of BACs led to abundance reductions of Firmicutes, Bacteroidetes, Acidobacteria and Chloroflexi, but Synergistetes was increased to 10.5%, which was almost not detected either in control or at a low level of BACs. And for dominant archaeal community, they tended to be shifted from acetotrophic to hydrogenotrophic methanogens with BACs increasing from low to high level. These findings provided some new insights for the role of BACs in anaerobic digestion, as well as resource recovery from WAS.

Occurrence, effect, and fate of residual microplastics in anaerobic digestion of waste activated sludge: A state-of-the-art review.

The plastic products have large consumption over last decades, resulting in a serious microplastics (MPs) pollution. Specially, the main removal way of MPs from wastewater is to transfer MPs from liquid to solid phase, leading to its enrichment in waste activated sludge (WAS). Anaerobic digestion has been served as the most potential technique to achieve both resource recovery and sludge reduction, herein this review provides current information on occurrence, effect, and fate of MPs in anaerobic digestion of WAS. The effects of MPs on WAS anaerobic digestion are greatly related to forms, particles sizes, contents, compositions and leachates of MPs. Also, the presence of MPs not only can change the effects of other pollutants on anaerobic digestion of WAS, but also can affect the fates of them. Besides, the future perspectives focused on the fate, effect and final removal of MPs during WAS anaerobic digestion process are outlined.

Response of anaerobic digestion of waste activated sludge to residual ferric ions.

This study was conducted to investigate the effects of residual ferric ions (FI), released from iron or its oxides for wastewater or waste activated sludge (WAS) treatment, on anaerobic digestion of WAS. Herein it was found that the anaerobic digestion process was greatly affected by FI dosages as well as FI distributions. The responses of performance and microorganism suggested that a low FI (e.g., 0.125 mmol/g volatile suspended solid (VSS)) enhanced methane production by 29.3%, and a medium FI (e.g., 0.3 mmol/g VSS) promoted short chain fatty acids accumulation to reach the maximum of 247 mg chemical oxygen demand /g VSS, conversely, a high FI (e.g., 0.9 mmol/g VSS) led to severe inhibition on acidogenesis and methanogenesis. The findings may provide some new insights for mechanism understanding on anaerobic digestion process influenced by iron or its oxides, as well as the disposal of WAS contained FI.

Total nitrogen removal in biochar amended non-aerated vertical flow constructed wetlands for secondary wastewater effluent with low C/N ratio: Microbial community structure and dissolved organic carbon release conditions.

Biochar was utilized to intensify constructed wetland (CW) for further organic and nitrogen removal from secondary wastewater. Four sets of non-aerated biochar amended vertical flow CW (VFCW) were developed to investigate the synergistic effects of biochar and microbes on pollutant removal. Results showed that the average COD and nitrogen removal efficiencies of VFCW1 (with 1% w/w biochar with microbe and plants) achieved 89.1 ± 5.6% and 90.2 ± 3.1% respectively, and their corresponding removal rates of 10.2 ± 0.8 mg-COD/(m3.d) and 3.57 ± 0.3 mg-TN/(m3.d) which were 35 and 52.3% higher than control. The biochar's dissolved organic carbon release in VFCWs indicated that water and acidic media portray the optimum conditions for nitrogen removal. The 16S RNA gene sequencing analysis indicated that in the biochar-amended VFCWs, bacterial phylum Proteobacteria (24.13-51.95%) followed by Chloroflexi (5.64-25.01%), Planctomycetes (8.48-14.43%), Acidobacteria (2.29-11.65%) were abundantly enhanced. Conclusively, incorporating biochar in non-aerated VFCWs is an efficient technique for enhancing nitrogen removal from secondary effluent.

Insights into palladium nanoparticles produced by Shewanella oneidensis MR-1: Roles of NADH dehydrogenases and hydrogenases.

Biologically synthesized palladium nanoparticles (bio-Pd) have attracted considerable interest as promising green catalysts for environmental remediation. However, the mechanisms by which microorganisms produce bio-Pd remain unclear. In the present study, we investigated the roles of Shewanella oneidensis MR-1 and its NADH dehydrogenases and hydrogenases (HydA and HyaB) in bio-Pd production using formate as the electron donor. The roles of NADH dehydrogenases and hydrogenases were studied by inhibiting NADH dehydrogenases and using hydrogenase mutants (ΔhydA, ΔhyaB, and ΔhydAΔhyaB), respectively. The results showed ~97% reduction of palladium by S. oneidensis MR-1 after 24 h using 250 µM palladium and 500 µM formate. Electron microscopy images showed the presence of bio-Pd on both the outer and cytoplasmic membranes of S. oneidensis MR-1. However, the inhibition of NADH dehydrogenases in S. oneidensis MR-1 resulted in only ~61% reduction of palladium after 24 h, and bio-Pd were not found on the outer membrane. The mutants lacking one or two hydrogenases removed 91-96% of palladium ions after 24 h and showed more cytoplasmic bio-Pd but less periplasmic bio-Pd. To the best of our knowledge, this is the first study to demonstrate the role of NADH dehydrogenases of S. oneidensis MR-1 in the formation of bio-Pd on the outer membrane. It also demonstrates that the hydrogenases (especially HyaB) of S. oneidensis MR-1 contribute to the formation of bio-Pd in the periplasmic space. This study provides mechanistic insights into the production of biogenic metal nanoparticles towards their possible use in industrial and environmental applications.

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.

Bioelectrochemical system for the enhancement of methane production by anaerobic digestion of alkaline pretreated sludge.

An increasing interest is devoted to combined microbial electrolysis cell-anaerobic digestion (MEC-AD) system which could convert waste activated sludge into biogas. In this study series tests were initially conducted to study the effect of alkaline pretreatment on AD system and the results showed that alkaline pretreatment could promote the dissolution of organic matters in the sludge and thus improve the methane production. Then, the methane production in combined MEC-AD system fed with alkaline-pretreated sludge was investigated. The results indicated that the methane productions increased by 37% and 42% when applied voltage was 0.5 V and 0.8 V. The microbial electrochemical system strongly promoted the growth of Euryarchaeota (Methanosaeta and Methanobacterium). Meanwhile, the abundance of Paraclostridium increased from 17.9% to 38.5% when applied voltage was 0.8 V, suggesting an enhanced fermentation and acetogenesis process. The results of energy balance estimation indicated that MEC-AD system at 0.5 V could achieve higher net energy output.

Electrochemistry-stimulated environmental bioremediation: Development of applicable modular electrode and system scale-up.

Bioelectrochemical systems (BESs) have been studied extensively during the past decades owing primarily to their versatility and potential in addressing the water-energy-resource nexus. In stark contrast to the significant advancements that have been made in developing innovative processes for pollution control and bioresource/bioenergy recovery, minimal progress has been achieved in demonstrating the feasibility of BESs in scaled-up applications. This lack of scaled-up demonstration could be ascribed to the absence of suitable electrode modules (EMs) engineered for large-scale application. In this study, we report a scalable composite-engineered EM (total volume of 1 m3), fabricated using graphite-coated stainless steel and carbon felt, that allows integrating BESs into mainstream wastewater treatment technologies. The cost-effectiveness and easy scalability of this EM provides a viable and clear path to facilitate the transition between the success of the lab studies and applications of BESs to solve multiple pressing environmental issues at full-scale.