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Microplastics as emerging pollutants have been heavily accumulated in the waste activated sludge (WAS) during biological wastewater treatment, which showed significantly diverse impacts on the subsequent anaerobic sludge digestion for methane production. However, a robust modeling approach for predicting and unveiling the complex effects of accumulated microplastics within WAS on methane production is still missing. In this study, four automated machine learning (AutoML) approach was applied to model the effects of microplastics on anaerobic digestion processes, and integrated explainable analysis was explored to reveal the relationships between key variables (e.g., concentration, type, and size of microplastics) and methane production. The results showed that the gradient boosting machine had better prediction performance (mean squared error (MSE) = 17.0) than common neural networks models (MSE = 58.0), demonstrating that the AutoML algorithms succeeded in predicting the methane production and could select the best machine learning model without human intervention. Explainable analysis results indicated that the variable of microplastic types was more important than the variable of microplastic diameter and concentration. The existence of polystyrene was associated with higher methane production, whereas increasing microplastic diameter and concentration both inhibited methane production. This work also provided a novel modeling approach for comprehensively understanding the complex effects of microplastics on methane production, which revealed the dependence relationships between methane production and key variables and may be served as a reference for optimizing operational adjustments in anaerobic digestion processes.

This work comparatively studied the different stress responses of anaerobic hydrogen-producing granular sludge (HPG) to several typical MPs in wastewater, i.e., polyethylene (PE), polyethylene terephthalate (PET), and polyvinyl chloride (PVC) MPs. A new approach to mitigating the inhibition caused by MPs based on biochar was then proposed. The results displayed that microbe in HPG had diverse tolerances to PE-MPs, PET-MPs and PVC-MPs, with the hydrogen production downgraded to 82.0 ± 3.2 %, 72.3 ± 2.5 % and 66.6 ± 2.3 % (p

The extensive application of anaerobic granular sludge (AGS) to wastewater treatment for methane recovery has drawn considerable attention to the system performances affected by the presence of emerging contaminants in wastewater such as nanoplastics. However, effective strategies on how to mitigate the inhibition caused by nanoplastics remained unavailable. In this study, a novel strategy using biochar to mitigate the inhibition on the AGS performances caused by polyethylene nanoplastics (PE-NPs) was proposed and the corresponding mitigating mechanisms involved were explored. The PE-NPs solely decreased the level of methane recovery of AGS to 71.3 ± 2.7% of control, which was subsequently increased to 85.6 ± 0.8% of control with the presences of both biochar and PE-NPs, although biochar solely showed no obvious effect on methane production. The addition of biochar also elevated the granule size of AGS, along with AGS integrity based on the morphological observation. Moreover, the distributions of live cells and functional microbes related to acidification and methanation increased with biochar addition compared to sole PE-NPs exposure. More extracellular polymeric substance (EPS) was secreted when biochar was involved in AGS systems, with more protein being detected to maintain the granule structure of AGS. Evaluation of adsorption tests indicated that biochar possessed stronger affinity for PE-NPs than AGS, thus capturing the PE-NPs that would originally contact AGS and posing less toxicity to microorganisms.

Micro(nano)plastics is an emerging contaminant in wastewater that has showed significant impacts on various biological treatment processes. Nevertheless, the underlying effects of micro(nano)plastics with different concentrations and sizes on the anaerobic hydrogen-producing granular sludge (HPG) were still unclear. This work firstly attempted to illustrate the microbial and physicochemical responses of HPG to a shock load of polyethylene microplastics (PE-MPs) with varied concentrations and sizes. The results revealed that the PE-MPs inhibitory effect on hydrogen production by HPG was both concentration- and size-dependent. Specifically, the increase of PE-MPs concentration and the decline of PE-MPs size to nano-sized plastics (NPs) significantly decreased the hydrogen yield, downgraded to 79.9 ± 2.6% and 63.0 ± 3.9% (p = 0.001, and 0.0002) of control, respectively, at higher MPs concentration and the smaller MPs size (i.e., NPs). The higher PE-MPs concentration and PE-NPs also suppressed extracellular polymeric substances (EPS) generation more severely. The critical bio-processes involved in hydrogen production were disturbed by PE-MPs, with the extent of negative impacts depending on the dosage and size of PE-MPs. These adverse impacts further manifested as granule disintegration and loss of cellular activity. Mechanism analysis highlighted the roles of oxidative stress, leachate released from PE-MPs, interaction between PE-NPs and granules inducing physical crushing of HPG that led to possible direct contact between cells and toxic substances.

Anaerobic hydrogen-producing granule (AHPG) has been successfully applied in hydrogen production from wastewater. While various types of microplastics in large amounts are readily detected in both municipal and industrial wastewaters, however, to date the response of AHPG to multiple coexisting microplastics in wastewater is unknown yet. Herein, this study provided a first insight into the acute exposure-response relationship between multiple coexisting microplastics and the AHPG during biological hydrogen production from wastewater. Fluorescence tagging found that many microplastics accumulated and covered on the surface of the whole granule. Morphology and particle size of microplastics-bearing AHPG were characterized by microscopic observation, showing that the shock load of microplastics in the wastewater at the studied concentrations (40 and 80 mg/L) made the granule loose and even break down with the decreased particle size. The visualization of extracellular polymeric substances (EPS) structure revealed that microplastics decreased EPS production by 8.8-16.7%. Microbial community analysis demonstrated that the acute exposure of microplastics did not drive the change in the microbial community diversity and composition. However, toxic leachates and upgraded oxidative stress induced by microplastics increased cell death up to 14.7% and decreased hydrogen production by 18.7%, when the AHPG exposed to 80 mg/L of microplastics. This work gained a new insight into the response of anaerobic microorganisms to coexisting microplastics in the real environment.

Excess sludge management is a restrictive factor for the development of municipal wastewater treatment plants. The addition of metabolic uncouplers has been proven to be effective in sludge reduction. However, the long-term effect of metabolic uncoupler o-chlorophenol (oCP) on the biological wastewater treatment system operated in anaerobic-oxic mode is still unclear. To this end, two parallel reactors operated in anaerobic-oxic mode with and without 10 mg/L of oCP addition were investigated for 91 days. The results showed that 56.1 ± 2.3% of sludge reduction was achieved in the oCP-added system, and the nitrogen and phosphorus removal ability were negatively affected. Dosing oCP stimulated the formation of microbial products and increased the DNA concentration, but resulted in a decrease in the electronic transport activity of activated sludge. Microbial community analysis further demonstrated that a significant reduction of bacterial richness and diversity occurred after oCP dosing. However, after stopping oCP addition, the pollutant removal ability of activated sludge was gradually increased, but the sludge yield, as well as species richness and diversity, did not recover to the previous level. This study will provide insightful guidance on the long-term application of metabolic uncouplers in the activated sludge system.

Polycyclic aromatic hydrocarbons (PAHs) are a group of common recalcitrant pollutant in industrial saline wastewater that raised significant concerns, whereas traditional activated sludge (AS) has limited tolerance to high salinity and PAHs toxicity, restricting its capacity to degrade PAHs. It is therefore urgent to develop a bioaugmented sludge (BS) system to aid in the effective degradation of these types of compounds under saline condition. In this study, a novel bioaugmentation strategy was developed by using halophilic Martelella sp. AD-3 for effectively augmented phenanthrene (PHE) degradation under 3% salinity. It was found that a 0.5∼1.5% (w/w) ratio of strain AD-3 to activated sludge was optimal for achieving high PHE degradation activity of the BS system with degradation rates reaching 2.2 mg⋅gVSS-1⋅h-1, nearly 25 times that of the AS system. Although 1-hydroxy-2-naphthoic acid (1H2N) was accumulated obviously, the mineralization of PHE was more complete in the BS system. Reads-based metagenomic coupled metatranscriptomic analysis revealed that the expression values of ndoB, encoding a dioxygenase associated with PHE ring-cleavage, was 5600-fold higher in the BS system than in the AS system. Metagenome assembly showed the members of the Corynebacterium and Alcaligenes genera were abundant in the strain AD-3 bioaugmented BS system with expression of 10.3±1.8% and 1.9±0.26%, respectively. Moreover, phdI and nahG accused for metabolism of 1H2N have been annotated in both above two genera. Degradation assays of intermediates of PHE confirmed that the activated sludge actually possessed considerable degradation capacity for downstream intermediates of PHE including 1H2N. The degradation capacity ratio of 1H2N to PHE was 87% in BS system, while it was 26% in strain AD-3. These results indicated that strain AD-3 contributed mainly in transforming PHE to 1H2N in BS system, while species in activated sludge utilized 1H2N as substrate to grow, thus establishing a syntrophic interaction with strain AD-3 and achieving the complete mineralization of PHE. Long-term continuous experiment confirmed a stable PHE removal efficiency of 93% and few 1H2N accumulation in BS SBR system. This study demonstrated an effective bioaugmented strategy for the bioremediation of saline wastewater containing PAHs.

This study demonstrated that Fe3O4 simultaneously improves the total production and formation rate of medium-chain fatty acids (MCFAs) and long-chain alcohols (LCAs) from waste activated sludge (WAS) in anaerobic fermentation. Results revealed that when Fe3O4 increased from 0 to 5 g/L, the maximal MCFA and LCA production increased significantly, and the optimal fermentation time was also remarkably shortened from 24 to 9 days. Moreover, Fe3O4 also enhanced WAS degradation, and the corresponding degradation rate in the fermentation system increased from 43.86 to 72.38% with an increase in Fe3O4 from 0 to 5 g/L. Further analysis showed that Fe3O4 promoted the microbe activities of all the bioprocesses (including hydrolysis, acidogenesis, and chain elongation processes) involved in the MCFA and LCA production from WAS. Microbial community analysis indicated that Fe3O4 increased the abundances of key microbes involved in abovementioned bioprocesses correspondingly. Mechanistic investigations showed that Fe3O4 increased the conductivity of the fermented sludge, providing a better conductive environment for the anaerobic microbes. The redox cycle of Fe(II) and Fe(III) existed in the fermentation system with Fe3O4, which was likely to act as electron shuttles to conduct electron transfer (ET) from the electron donor to the acceptor, thus increasing ET efficiency. This study provides an effective method for enhancing the biotransformation of WAS into high-value products, potentially bringing economic benefits to WAS treatment.

This study describes an integrated granular sludge and fixed-biofilm (iGB) reactor innovatively designed to carry out the anammox/partial-denitrification (A/PD) process for nitrogen removal with mainstream municipal wastewater. The iGB-A/PD reactor consists of anammox granules inoculated in the lower region of reactor and an acclimated fixed-biofilm positioned in the upper region. Compared to the other reported A/PD systems for mainstream wastewater treatment, this iGB-A/PD reactor is notable due to its higher quality effluent with a total inorganic nitrogen (TIN) of ∼3 mg•L-1 and operation at a high nitrogen removal rate (NRR) of 0.8 ± 0.1 kg-N•m-3•d-1. Reads-based metatranscriptomic analysis found that the expression values of hzsA and hdh, key genes associated with anammox, were much higher than other functional genes on nitrogen conversion, confirming the major roles of the anammox bacteria in nitrogen bio-removal. In both regions of the reactor, the nitrate reduction genes (napA/narG) had expression values of 56-99 RPM, which were similar to that of the nitrite reduction genes (nirS/nirK). The expression reads from genes for dissimilatory nitrate reduction to ammonium (DNRA), nrfA and nirB, were unexpectedly high, and were over the half of the levels of reads from genes required for nitrate reduction. Kinetic assays confirmed that the granules had an anammox activity of 16.2 g-NH4+-N•kg-1-VSS•d-1 and a nitrate reduction activity of 4.1 g-N•kg-1-VSS•d-1. While these values were changed to be 4.9 g- NH4+-N•kg-1-VSS•d-1and 4.3 g-N•kg-1-VSS•d-1 respectively in the fixed-biofilm. Mass flux determination found that PD and DNRA was responsible for ∼50% and ∼25% of nitrate reduction, respectively, in the whole reactor, consistent with high effluent quality and treatment efficiency via a nitrite loop. Metagenomic binning analysis revealed that new and unidentified anammox species, affiliated with Candidatus Brocadia, were the dominant anammox organisms. Myxococcota and Planctomycetota were the principal organisms associated with the PD and DNRA processes, respectively.

Methane recovery from waste activated sludge (WAS) through anaerobic digestion is generally restricted by the poor degradability of WAS. Herein, a novel sludge pretreatment technology by using the calcium hypochlorite (Ca(ClO)2) in enhancing the methane production from WAS anaerobic digestion was reported. The solubilization of WAS was significantly increased after 10-240 mg Ca(ClO)2/g VS (VS: volatile solids) pretreatment for 48 h, under which the solubilization was 1.7-3.4 folds (i.e., 0.17-0.34 mg SCOD/mg VS; SCOD: soluble chemical oxygen demand) higher than that without Ca(ClO)2 pretreatment (i.e., 0.1 mg SCOD/mg VS). Correspondingly, the methane production was increased from 250.0 ± 5.3 mL/g VS to 385.1 ± 3.3 mL/g VS with the doses of Ca(ClO)2 increasing from 10 mg/g VS to 240 mg/g VS, resulted in an increasing methane production of 3.6%-59.7% than that without Ca(ClO)2 pretreatment. The microbial community composition results exhibited that the populations of key acidogens (e.g., Longilinea sp.) and methanogens (e.g., Methanosaeta sp.) were both reduced significantly. Moreover, Ca(ClO)2 decreased the cells viability, leading to a 76.2% reduction of living cells fraction. Accordingly, it was further confirmed that high dosage of Ca(ClO)2 could inhibit three microbial-related processes relevant to methane production, i.e., acidification, hydrolysis and methanogenesis.

This work proved an efficient method to significantly increase methane production from anaerobic digestion of WAS. This method is to reflux proper of digestion liquid into waste activated sludge pretreatment unit (pH 9.5 for 24 h). The yield of maximum methane improved between 174.2 ± 7.3 and 282.5 ± 14.1 mL/g VSS with the reflux ratio of digestion liquid increasing from 0% to 20%. It was observed that the biodegradable organics in the digestion liquid did not affect the biological processes related to anaerobic digestion but increased methane production through reutilization. The ammonium in the digestion liquid was the main contributor to the increase in methane production via promoting sludge solubilization, but refractory organics were the major inhibitors to anaerobic digestion. It should be emphasized that the metal ions present in the digestion liquid were beneficial rather than harmful to the biological processes in the anaerobic digestion, which may be connected with the fact that certain metal ions were involved in the expression and activation of key enzymes. In addition, it was found that anaerobes in digestion liquid were another potential contributor to the enhanced anaerobic digestion.

In this work, a rhamnolipid (RL) pretreatment technology was proposed to promote methane production from two-phase anaerobic digestion of waste activated sludge. In the first phase (i.e., acidogenic phase), the WAS hydrolysis and acidogenesis were significantly enhanced after RL pretreatment for 4 day, under which the concentration of soluble protein and the short-chain fatty acids (SCFA) in the presence of RL at 0.04 g/g TSS was respectively 2.50 and 5.02 times higher than that without RL pretreatment. However, methane production was inhibited in the presence of RL. In the second phase (i.e., methanogenic phase), batch biochemical methane potential tests suggested that the addition of RL is effective in promoting anaerobic methane production. With an increase of RL dosage from 0 to 0.04 g/g TSS, the cumulative methane yield increased from 100.42 ± 3.01 to 168.90 ± 5.42 mL. Although the added RL could be utilized to produce methane, it was not the major contributor to the enhancement of methane yield. Further analysis revealed that total cumulative yield from the entire two-phase anaerobic digestion (sum of the yield of the acidogenic phase and methanogenic phase) increased from 113.42 ± 3.56 to 164.18 ± 5.20 mL when RL dosage increased from 0 to 0.03 g/g TSS, indicating that the addition of RL induced positive effect on the methane production of the entire two-phase anaerobic digestion. The enzyme activity analysis showed that although higher dosages of RL still inhibited the microorganisms related to methanogenesis to some extends in the methanogenic phase, the inhibitory effect was significantly weakened compared to the acidogenic phase. Microbial analysis revealed that RL reduced the abundance of Candidatus_Methanofastidiosum sp. while increased the abundance of Methanosaeta sp., causing the major methanogenesis pathway to change from hydrogenotrophic to aceticlastic. Moreover, the community of hydrolytic microbes and acidogens was shifted in the direction that is conducive to hydrolysis-acidogenesis. The findings reported not only expand the application field of RL, but also may provide supports for sustainable operation of wastewater treatment plants (WWTPs).

Due to the environmental risks caused by microplastics, understanding the sources and characteristics of microplastics and cutting off their routes into the environment are crucial. However, so far, studies on microplastics in the landfill leachate system (a major pathway of microplastics into the environment) are still limited, especially for tiny particles

A novel zerovalent iron (ZVI) technique to simultaneously improve the production of medium-chain fatty acids (MCFAs) from waste activated sludge (WAS) and enhance WAS degradation during anaerobic WAS fermentation was proposed in this study. Experimental results showed that the production and selectivity of MCFAs were effectively promoted when ZVI was added at 1-20 g/L. The maximum MCFAs production of 15.4 g COD (Chemical Oxygen Demand)/L and MCFAs selectivity of 71.7% were both achieved at 20 g/L ZVI, being 5.3 and 4.8 times that without ZVI (2.9 g COD/L and 14.9%). Additionally, ZVI also promoted WAS degradation, which increased from 0.61 to 0.96 g COD/g VS when ZVI increased from 0 to 20 g/L. The microbial community analysis revealed that the ZVI increased the populations of key anaerobes related to hydrolysis, acidification, and chain elongation. Correspondingly, the solubilization, hydrolysis, and acidification processes of WAS were revealed to be improved by ZVI, thereby providing more substrates (short-chain fatty acids (SCFAs)) for producing MCFAs. The mechanism studies showed that ZVI declined the oxidation-reduction potential (ORP), creating a more favorable environment for the anaerobic biological processes. More importantly, ZVI with strong conductivity could act as an electron shuttle, contributing to increasing electron transfer efficiency from electron donor to acceptor. This strategy provides a new paradigm of transforming waste sludge into assets by a low-cost waste to bring significant economic benefits to sludge disposal and wastewater treatment.

Traditional bioenergy recovery in the form of short chain fatty acids (SCFAs) from waste activated sludge (WAS) is generally limited by economic unattractiveness and complexity of products separation. Herein, a novel biotechnology process of two-stage anaerobic fermentation for converting the WAS into high energy density, easy-separated medium chain fatty acids (MCFAs) and long-chain alcohols (LCAs) was evaluated. In this process, the WAS was first converted to WAS alkaline fermentation liquid (WASAFL), serving as electron acceptors (EAs) and inoculum, then adding ethanol as electron donor (ED) for chain elongation (CE). The co-production of MCFAs and LCAs during CE were studied under three different ED to EA ratios, i.e., 3:1, 4:1 and 5:1. Experimental results demonstrated that when the ratio of ED to EA increased from 3:1 to 5:1, the production of MCFA and LCAs respectively increased from 5.57 ± 0.17 and 2.58 ± 0.18 to7.67 ± 0.48 and 4.21 ± 0.19 g COD/L. A similar observation was made in the total product electron efficiency, increasing from 59.9% to 72.1%. However, the highest total product selectivity (i.e., 68.0%) and highest products production yield (i.e., 59.77%) were not achieved at the ED to EA ratio of 5:1 due to toxicity caused by higher accumulation of n-caproate. The kinetic analysis further confirmed that high ratio of ED to EA induced improvement in product maximum yield, production rate for both MCFAs and LCAs. Microbial community analysis indicated that Clostridium, Caproiciproducens, Acinetobacter, Exilispira, and Oscillibacter were clearly enriched in the CE reactor and had positive correlation with MCFAs and LCAs production.

Polyvinyl chloride microplastics (PVC-MPs) are emerging contaminants affecting biological wastewater treatment processes. However, most of the previous studies mainly focused on their short-term impacts on floc sludge, with little work being conducted to explore their potential effects on more complex anaerobic granular sludge (AGS), which has been widely used for high-strength organic wastewater treatment. In this paper, the long-term effects of PVC-MPs on AGS were investigated via continuous feeding tests that are representative of real wastewater treatment processes. The results of a continuous 264 days test showed that the prolonged exposure of PVC-MPs at 15-150 MPs·L-1 significantly (p = 7.86 × 10-37, 3.44 × 10-43, and 5.29 × 10-46) inhibited the COD removal efficiency of AGS by 13.2%-35.5%, accompanied by a 11.0%-32.3% decreased production of methane and 40.3%-272.7% increased accumulation of short-chain fatty acids (SCFAs). In addition, the PVC-MPs exposure suppressed the secretion of extracellular polymeric substances (EPS), causing AGS and the inside microorganisms to lose the protection of EPS, thereby resulting in granule breakage and decreased cells viability. Aligning with the deteriorated performance, the long-term exposure of PVC-MPs reduced the total microbial populations and the relative abundances of key methanogens and acidogens. A toxicity mechanism assessment revealed that the negative impacts induced by PVC-MPs are mainly attributed to the toxic leachate and excess oxidative stress.

The negative effects of ubiquitous microplastics on wastewater treatment have attracted increasing attention. However, the potential impacts of microplastics on anaerobic granular sludge (AGS) remain unknown. To fill this knowledge gap, this paper investigated the response of AGS to the exposure of model microplastics (polyethylene terephthalate (PET-MPs)) and provided insights into the mechanisms involved. The 84 days' long-term exposure experiments demonstrated that PET-MPs, at relatively low level (15 MP L-1) did not affect AGS performance during anaerobic wastewater treatment, while 75-300 MP L-1 of PET-MPs caused the decreases of COD removal efficiency and methane yields by 17.4-30.4% and 17.2-28.4%, accompanied with the 119.4-227.8% increase in short-chain fatty acid (SCFA) accumulation and particle breakage. Extracellular polymeric substances (EPS) analysis showed that dosage-dependent tolerance of AGS to PET-MPs was attributed to the induced EPS producing protection role, but PET-MPs at higher concentrations (75-300 MP L-1) suppressed EPS generation. Correspondingly, microbial community analysis revealed that the populations of key acidogens (e.g., Levilinea sp.) and methanogens (e.g., Methanosaeta sp.) decreased after long-term exposure to PET-MPs. Assessment of the toxicity of PET-MPs revealed that the leached di-n-butyl phthalate (DBP) and the induced reactive oxygen species (ROS) by PET-MPs were causing toxicity towards AGS, confirmed by the increases in cell mortality and lactate dehydrogenase (LDH) release. These results provide novel insights into the ecological risk assessment of microplastics in anaerobic wastewater treatment system.

In this work, Fe(II) catalyzing sodium percarbonate (Fe(II)/SPC) was managed to facilitate waste activated sludge (WAS) dewatering for the first time. The results showed that after WAS was treated by 20 mg/g total suspended solids (TSS) Fe(II) and 50 mg/g TSS SPC, the water content of sludge cake (WCSC) by press filtration and capillary suction time (CST) dropped from 90.8% ± 1.6% and 96.1 ± 4.0 s (the control) to 55.6% ± 1.4% and 30.1 ± 2.5 s, respectively. The mechanism investigations indicated that four intermediates or products (i.e., •OH, H2O2, Fe(II), and Fe(III)) generated in the Fe(II)/SPC process were responsible for the improved WAS dewaterability, and •OH and Fe(III) were the two major contributors. It was found that •OH collapsed and fragmented extracellular polymeric substances, damaged cell wall and permeabilized cytoplasmic membrane, and transformed conformation of the extracellular proteins secondary structure via both affecting the hydrogen bond maintaining α-helix and cracking disulfide bond in cysteine residues while Fe(III), the oxidization product of Fe(II), decreased the surface electronegativity and water-affinity surface areas of WAS flocs. As a result, the bound water release, flocculability, surface hydrophobicity, drain capability, and flowability of WAS flocs were strengthened whereas the compact surface structure, colloidal forces, network strength, gel-like structure, and apparent viscosity of WAS flocs were weakened. In addition, Fe(II)/SPC process also reduced the recalcitrant organics and fecal coliforms in sludge, which facilitated land application of dewatered sludge. The findings acquired in this work not only deepens our understanding of Fe(II)/SPC-involved WAS treatment process but also may guide engineers to develop both effective and promising strategies to better condition WAS for dewatering in the future.

Wastewater treatment plants (WWTPs) have been identified as one of the reservoirs of antibiotics. Although nitrifying bacteria have been reported to be capable of degrading various antibiotics, there are very few studies investigating long-term effects of antibiotics on kinetic and microbial responses of nitrifying bacteria. In this study, cephalexin (CFX) and sulfadiazine (SDZ) were selected to assess chronic impacts on nitrifying sludge with stepwise increasing concentrations in two independent bioreactors. The results showed that CFX and SDZ at an initial concentration of 100 μg/L could be efficiently removed by enriched nitrifying sludge, as evidenced by removal efficiencies of more than 88% and 85%, respectively. Ammonia-oxidizing bacteria (AOB) made a major contribution to the biodegradation of CFX and SDZ via cometabolism, compared to limited contributions from heterotrophic bacteria and nitrite-oxidizing bacteria. Chronic exposure to CFX (≥30 μg/L) could stimulate ammonium oxidation activity in terms of a significant enhancement of ammonium oxidation rate (p

The increasing use of perfluorooctanoic acid (PFOA) raises concerns about its potential toxicity to the environment. However, the interaction between PFOA and aerobic granular sludge has never been documented. This work therefore aims to provide such support through investigating the fate of PFOA at environmentally relevant levels in aerobic granular sludge systems and its impact on aerobic granular sludge. Experimental results showed that 32.0%∼36.4% of wastewater PFOA was removed by aerobic granular sludge in stable operation when PFOA concentration was ranged from 0.1 to 1.0 mg/L. Mass balance analyses and X-ray photoelectron spectroscopy survey scan revealed that the removal of PFOA was dominated by adsorption rather than biodegradation, and sorption kinetic analysis indicated that inhomogeneous multilayer adsorption was responsible for this removal. The adsorbed PFOA deteriorated the settleability of granular sludge and biological nitrogen and phosphorus removal significantly. Experimental results showed that 1.0 mg/L PFOA inhibited anaerobic phosphate release, aerobic phosphate uptake, nitrate reduction, and nitrite reduction processes by 60%, 50%, 13.1%, and 5.8%, respectively. It was observed that PFOA induced large amounts of filamentous villus growing on the surface and increased the extracellular polymeric substances of granular sludge. Fourier-transform infrared spectra and X-ray photoelectron spectroscopy spectrum showed that several function groups in extracellular polymeric substances such as hydroxyl groups, amides and polysaccharides were affected by PFOA. It was also found that PFOA inhibited the cyclic transformations of polyhydroxyalkanoates and glycogen. Microbial community analyses showed that PFOA decreased the abundances of Nitrosomonas, Nitrospira, Accumulibacter, and other function microbes such as Rhodospirillaceae, Thauera, and Azoarcus.

Effective sludge treatment with bioenergy production is attracting increasing interests as large quantities of waste activated sludge (WAS) are produced during the wastewater treatment. In this study, a new biotechnical process for converting the WAS alkaline fermentation liquor (WASAFL) into valuable, easy-separated medium chain fatty acids (MCFAs) through chain elongation (CE) was investigated, which may provide a new insight into sludge treatment. In the process, ethanol was served as the electron donor (EDs) and WASAFL were main electron acceptors (EAs). The MCFAs productions were investigated under three different ED to EA ratios (i.e., 1:2, 1:1 and 2:1). The result showed that MCFAs production was increased from 2.88 ± 0.01 to 5.28 ± 0.18 g COD/L with the increase of ED to EA ratio. However, the highest MCFA selectivity was achieved at 72.9% when the ED to EA ratio was 1:1. The decrease in the selectivity at high ED:EA ratio is mainly due to the production of higher alcohol (i.e., n-butanol and n-hexanol). The thermodynamic analysis confirmed all CE processes for MCFAs production from WASAFL were exothermic reactions, with the spontaneity and energy release of the reactions increased with the ethanol level. The microbial community analysis showed that the relative abundances of Clostridium, Oscillibacter, Leptolinea and Exilispira were positively correlated with the MCFAs production. The metagenomic analysis suggested that both the reverse β-oxidization pathway and fatty acid biosynthesis pathway contributed to the CE process in the studied system. The functional enzymes were mainly associated within Clostridium, with Clostridium Kluyveri, Clostridium botulinum and Clostridium magnum being likely the key species responsible for the CE process.

The macrolide antibiotic roxithromycin is widely detected in varying aquatic environments, especially in the wastewater systems, as an emerging contaminant and leads to significant impacts on the microorganisms involved. In this study, the impact of a shock load of roxithromycin on waste activated sludge (WAS) anaerobic digestion was comprehensively investigated. The biochemical methane potential tests showed that the methane production from WAS anaerobic digestion was significantly inhibited by roxithromycin. With the dosage of roxithromycin increasing from 0 to 1000 μg/L, the maximum cumulative methane production decreased from 163.5 ± 2.6 mL/g VS to 150.9 ± 4.5 mL/g VS. In particular, roxithromycin inhibited the acidogenesis and methanogenesis in WAS anaerobic digestion, leading to the decreased methane production. The methanogenic archaea in the studied system mainly belonged to the genera of Methanoseata, Candidatus Methanofastidiosum and Methanolinea and their relative abundances also decreased with roxithromycin addition. The analysis of antibiotic resistance genes (ARGs) in the digested sludge indicated that the abundances of most ARGs detected in this study were increased with roxithromycin exposure, suggesting the potential of growing antibiotic resistance, which was probably caused by enhancing the effect of esterases, methylases and phosphorylases. This work reveals how roxithromycin affects the WAS anaerobic digestion and the change of ARGs in the anaerobic digestion with roxithromycin exposure, and provides useful information for practical operation.

© 2018 Elsevier Ltd Antibiotic wastewater has become a major concern due to the toxicity and recalcitrance of antibiotics. Anaerobic membrane bioreactors (AnMBRs) are considered alternative technology for treating antibiotic wastewater because of their advantages over the conventional anaerobic processes and aerobic MBRs. However, membrane fouling remains the most challenging issue in the AnMBRs’ operation and this limits their application. This review critically discusses: (i) antibiotics removal and antibiotic resistance genes (ARGs) in different types of AnMBRs and the impact of antibiotics on membrane fouling and (ii) the integrated AnMBRs systems for fouling control and removal of antibiotics. The presence of antibiotics in AnMBRs could aggravate membrane fouling by influencing fouling-related factors (i.e., sludge particle size, extracellular polymeric substances (EPS), soluble microbial products (SMP), and fouling-related microbial communities). Conclusively, integrated AnMBR systems can be a practical technology for antibiotic wastewater treatment.

Microbial internal storage processes have been demonstrated to occur and play an important role in activated sludge systems under both aerobic and anoxic conditions when operating under dynamic conditions. High-rate anaerobic reactors are often operated at a high volumetric organic loading and a relatively dynamic profile, with large amounts of fermentable substrates. These dynamic operating conditions and high catabolic energy availability might also facilitate the formation of internal storage polymers by anaerobic microorganisms. However, so far information about storage under anaerobic conditions (e.g., anaerobic fermentation) as well as its consideration in anaerobic process modeling (e.g., IWA Anaerobic Digestion Model No. 1, ADM1) is still sparse. In this work, the accumulation of storage polymers during anaerobic fermentation was evaluated by batch experiments using anaerobic methanogenic sludge and based on mass balance analysis of carbon transformation. A new mathematical model was developed to describe microbial storage in anaerobic systems. The model was calibrated and validated by using independent data sets from two different anaerobic systems, with significant storage observed, and effectively simulated in both systems. The inclusion of the new anaerobic storage processes in the developed model allows for more successful simulation of transients due to lower accumulation of volatile fatty acids (correction for the overestimation of volatile fatty acids), which mitigates pH fluctuations. Current models such as the ADM1 cannot effectively simulate these dynamics due to a lack of anaerobic storage mechanisms.

Dissolved oxygen (DO) is commonly recognized as an important factor influencing nitrous oxide (N2O) production by ammonia-oxidizing bacteria (AOB). However, it has been difficult to separate the true effect of DO from that of nitrite, as DO variation often affects nitrite accumulation. The effect of DO on N2O production by an enriched nitrifying sludge, consisting of both AOB and nitrite-oxidizing bacteria (NOB), was investigated in this study. Nitrite accumulation was minimised by augmenting nitrite oxidation through the addition of an enriched NOB sludge. It was demonstrated that the specific N2O production rate increased from 0 to 1.9 ± 0.09 (n = 3) mg N2O-N/hr/g VSS with an increase of DO concentration from 0 to 3.0 mg O2/L, whereas N2O emission factor (the ratio between N2O nitrogen emitted and the ammonium nitrogen converted) decreased from 10.6 ± 1.7% (n = 3) at DO = 0.2 mg O2/L to 2.4 ± 0.1% (n = 3) at DO = 3.0 mg O2/L. The site preference measurements indicated that both the AOB denitrification and hydroxylamine (NH2OH) oxidation pathways contributed to N2O production, and DO had an important effect on the relative contributions of the two pathways. This finding is supported by analysis of the process data using an N2O model describing both pathways. As DO increased from 0.2 to 3.0 mg O2/L, the contribution of AOB denitrification decreased from 92% - 95%-66% - 73%, accompanied by a corresponding increase in the contribution by the NH2OH oxidation pathway.

Aerobic digestion is considered to be a common process for the stabilization of waste activated sludge (WAS) in the small-sized wastewater treatment systems, while the broad application of titanium dioxide nanoparticles (TiO2 NPs) results in their unavoidable existence in WAS aerobic digestion, with its role in aerobic sludge digestion being never documented. This study set up a series of aerobic sludge digesters to evaluate the previously unknown role of TiO2 NPs on the performance of the digesters. The volatile solids (VS) degradation percentage increased from 21.9 ± 0.6% to 26.9 ± 0.1% − 30.0 ± 0.3% with the different contents of TiO2 NPs (0, 1, 20 and 50 mg/L). Similarly, the total inorganic nitrogen production increased from 23.1 ± 0.3 to 31.0 ± 0.1 mg N/g VS with the rising TiO2 NPs concentrations from 0 to 50 mg/L. The microbial analysis suggested that TiO2 NPs contributed to the accumulation of specific microbes correlated with the degradation of organic substances and the conversion of nitrogen compounds. Model-based analysis showed the higher biodegradability and hydrolysis rate of sludge with TiO2 NPs. Further mechanistic studies indicated that the enhancement of WAS solubilization and the degradation of recalcitrant substances (e.g., humic acid and cellulose) contributed to the better performance of experimental aerobic digesters, which was confirmed by the fourier transformation infrared spectroscopy (FTIR) indicating the converting of these materials into biodegradable substrates for digestion with TiO2 NPs. It could be inferred from this investigation that aerobic digestion rather than anaerobic digestion would be a more suitable treatment method for sludge containing TiO2 NPs.

This paper presents a modelling study aimed at minimizing the environmental foot print of a membrane bioreactor (MBR) for wastewater treatment. Specifically, an integrated model for MBR was employed in view of the management optimization of an MBR biological nutrient removal (BNR) pilot plant in terms of operational costs and direct greenhouse gases emissions. The influence of the operational parameters (OPs) on performance indicators (PIs) was investigated by adopting the Extended-FAST sensitivity analysis method. Further, a multi-objective analysis was performed by applying the Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS). The results show-up that the sludge retention time is the OP mostly affecting all the investigated PIs. By applying the set of optimal OPs, there was a reduction of 48% and 10% of the operational costs and direct emissions, respectively.