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With increased industrial development, vast heavy metals are inevitably discharged into wastewater. Cu 2+ is one of the most hazardous heavy metals in biotreatment. However, the potential effect of Cu 2+ on denitrifying granular sludge is still unknown. This work assesses the response of denitrifying granular sludge to Cu 2+ stress from multiple aspects. The denitrifying granular sludge could tolerate 5 mg L -1 Cu 2+ , while the nitrogen removal efficiency decreased to 48.5% under 10 mg L -1 Cu 2+ . Enzyme activity and carbohydrate metabolism were inhibited, and the denitrifying bacteria were washed out under Cu 2+ stress. The resulting deteriorated state was reversed by phosphate. The nitrogen removal efficiency recovered to 99% after 10 days, and the enzyme activity also recovered to the original level. Membrane transport, transcription and cellular processes were promoted. Overall, the results of this work provide a feasible strategy to rapidly restore the metabolic activity of denitrifying granular sludge under Cu 2+ stress., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Antibiotics are widely found in nitrogen-containing wastewater, which may affect the operation stability of anaerobic ammonium oxidation (anammox)-based biological treatment systems. Extracellular polymeric substances (EPSs) of anammox sludge play a pivotal role in combining with antibiotics; however, the exact role and how the structure of the leading component of EPSs (i.e., extracellular proteins) changes under antibiotic stress remain to be elucidated. Here, the interaction between sulfamethoxazole and the extracellular proteins of anammox sludge was investigated via multiple spectra and molecular simulation. Results showed that sulfamethoxazole statically quenched the fluorescent components of EPSs, and the quenching constant of the aromatic proteins was the largest, with a value of 1.73 × 10 4 M -1 . The overall binding was an enthalpy-driven process, with Δ H = -75.15 kJ mol -1 , Δ S = -0.175 kJ mol -1 K -1 , and Δ G = -21.10 kJ mol -1 at 35 °C. The O-P-O and C═O groups responded first under the disturbance of sulfamethoxazole. Excessive sulfamethoxazole (20 mg L -1 ) would decrease the ratio of α-helix/(β-sheet + random coil) of extracellular proteins, resulting in a loose structure. Molecular docking and dynamic simulation revealed that extracellular proteins would provide abundant sites to bind with sulfamethoxazole, through hydrogen bond and Pi-Akyl hydrophobic interaction forces. Once sulfamethoxazole penetrates into the cell surface and combines with the transmembrane ammonium transport domain, it may inhibit the NH 4 + transport. Our findings enhance the understanding on the interaction of extracellular proteins and sulfamethoxazole, which may be valuable for deciphering the response property of anammox sludge under the antibiotic stress.

The arsenic in livestock wastewater would induce adverse impact on the biological treatment technology such as anaerobic ammonium oxidation (anammox) process. Extracellular polymeric substances (EPS) play an important role in resisting such toxicity. Unfortunately, the role of EPS in protecting anammox from As(III) and the mechanisms underlying the protection still remains unclear. This work comprehensively evaluated the acute toxicity of arsenic on anammox sludge and investigated the binding property and interaction mechanism. The results revealed that the half maximal inhibitory concentration (IC 50 ) of As(III) on anammox sludge was estimated to be 408 mg L -1 , which decreased to 41.97 mg L -1 when EPS was exfoliated. Complexation and hydrophobic interactions were the leading forces in preventing arsenic invasion. Protein was the main component that complexes with As(III), and O-H, -NH, -CO were binding sites. The response sequence of organic component in EPS to As(III) was ordered as hydrocarbons-proteins-polysaccharides-aliphatic amines., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The increase of emerging contaminants, such as surfactants, is one of the major challenges to biological wastewater treatment. However, the potential impact of linear alkylbenzene sulphonates (LAS), a major class of anionic surfactants, on anammox process is unclear. The long-term effects of sodium dodecyl benzene sulfonate (SDBS, as a model LAS) on reactor performance, microbial community and sludge properties were investigated in this study. The presence of 5 mg L -1 SDBS promoted the release of extracellular microbial products from anammox granules and the wash-out of anammox population via effluent. Despite sludge disaggregation, the reactor performance was robust to the exposure of 5 mg L -1 SDBS due to functional redundancy. With the further increase of SDBS to 10 mg L -1 , the metabolic activity of anammox biomass and the transcription and post-translation of hydrazine dehydrogenase were significantly decreased. The potential mechanism might be associated with the damage on cell membrane that induced the leakage of intracellular matrix. These results highlight the need to consider the potential risk of LAS to operation of anammox process in biological wastewater treatment plant., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The widespread use of copper nanoparticles (CuNPs) has attracted increasing concern because of their potential effects on biological wastewater treatment. However, their effect on granule-based denitrification systems is unclear. Hence, the effects of CuNPs on denitrifying granules were investigated during long-term operation. The results showed that 51.9% of nitrogen removal capacity was lost after exposure to 5 mg L -1 CuNPs, with the amount of Cu(II) gradually increasing with elevating CuNP levels. Moreover, the relative abundance of denitrifying bacteria (Castellaniella) and denitrifying functional genes (nirK, napA, narG and nosZ) obviously decreased. Meanwhile, the specific denitrification activity, the content of extracellular polymeric substances and dehydrogenase activity decreased by 44.0%, 15.2% and 99.9%, respectively, compared to their values in the initial sludge. Considering the downtrend in the abundance of copper resistance genes, it was deduced that the toxicity of CuNPs was mainly or at least partially due to the release of Cu(II)., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

High-rate denitrification is a popular and efficient process for treatment of nitrate-rich wastewater. Knowing the effect of heavy metals on denitrification is essential for industrial development. In the present study, the long-term impacts of Zn(II) on denitrifying granular sludge were investigated. The suppression threshold of Zn(II) on denitrifying bacteria was 10 mg L -1 for long-term exposure. The nitrogen removal rate was decreased by long-term addition of 10 mg L -1 Zn(II). Castellaniella and Klebsiella were the two dominant genera under Zn(II) stress. The relative abundance of Klebsiella sharply decreased to 4.64% after the addition of 10 mg L -1 Zn(II), whereas Castellaniella was susceptible to 2 mg L -1 Zn(II), revealing that Castellaniella mainly was devoted to denitrification under no or low Zn(II) stress conditions, whereas Klebsiella was effective under high Zn(II) stress., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The increasing use of engineered Ag nanoparticles (NPs) in consumer products raises great concerns about their environmental impacts, but their potential impact on anaerobic ammonium oxidation (anammox) bacteria in wastewater treatment remains unclear. In this study, the response of anammox consortia inhabiting in granules to long-term exposure of AgNPs was investigated. The gradually increased stress of AgNPs (1, 10, 50 mg L -1 ) showed no adverse effects on the nitrogen removal performance and on the relative abundance of Ca. Kuenenia inhabiting in granules. Notably, in response to the escalating burden of AgNPs, anammox consortia tended to secrete more protein but less polysaccharide, and meanwhile increase the relative abundances of Ag + efflux protein SilA and regulatory protein SilR. Overall, these results suggested that the strong resistance of anammox granules to AgNPs might be mainly attributed to the defense role of extracellular polymeric substances and gene-based regulation of ion transport., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Magnetic nanoparticles (NPs) have been widely applied in environmental remediation, biomass immobilization and wastewater treatment, but their potential impact on anaerobic ammonium oxidation (anammox) biomass remains unknown. In this study, the short-term and long-term impacts of maghemite NPs (MHNPs) on the flocculent sludge wasted from a high-rate anammox reactor were investigated. Batch assays showed that the presence of MHNPs up to 200 mg L -1 did not affect anammox activity, reactive oxygen species production, or cell membrane integrity. Moreover, long-term addition of 1-200 mg L -1 MHNPs had no adverse effects on reactor performance. Notably, the specific anammox activity, the abundance of hydrazine synthase structural genes and the content of extracellular polymeric substance were increased with elevated MHNP concentrations. Meanwhile, the community structure was shifted to higher abundance of Candidatus Kuenenia indicated by high-throughput sequencing. Therefore, MHNPs could be applied to enhance anammox flocculent sludge due to their favorable biocompatibility., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Given the increasing use of nanoscale zero-valent iron (NZVI) particles for environmental remediation and wastewater treatment, their potential impact on anaerobic ammonium oxidation (anammox) bacteria was investigated in this study using anammox sludge. Batch assays showed that NZVI concentrations up to 200mgL -1 did not affect anammox activity, reactive oxygen species production, and cell membrane integrity. The nitrogen removal efficiency of the continuous-flow reactor fluctuated in the presence of 20 or 50mgL -1 NZVI, but it could return to normal over time, even at 200mgL -1 NZVI. 16S rDNA-based high-throughput sequencing indicated that although the presence of 10, 20, 50, and 200mgL -1 NZVI to some extent affected microbial composition, the anammox bacteria (Candidatus Kuenenia) never lost its dominance. The abundance of gene families that are related to the assimilation and utilization of iron was down-regulated in response to the stress of high-level NZVI., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The increasing application of engineered nanoparticles (NPs) has posed an emerging challenge to wastewater treatment processes. The short-term impacts of CuNPs, CuONPs, ZnONPs and AgNPs on anaerobic ammonium oxidation (anammox) process were investigated firstly in this study. CuONPs, ZnONPs and AgNPs up to 50mgg -1 suspended solid (SS) did not affect anammox activity, reactive oxygen species (ROS) production or cell membrane integrity. However, 1.25mgg -1 SS CuNPs significantly inhibited the anammox activity and the loads that caused 50% inhibition were 4.64±1.24 and 3.27±0.79mgg -1 SS for anammox granules and flocs, respectively. 5mgg -1 SS CuNPs caused serious accumulation of the toxic intermediate N 2 H 4 . Furthermore, CuNPs interacted with extracellular polymeric substances by specifically bonding to tyrosine or tryptophan-containing groups, C-O-C in polysaccharides and -OH in polymeric compounds. Therefore, this study calls for more attention to the risks of NPs to the anammox-based processes., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, the behavior, distribution and form dynamics of overloaded Cu(II) in anaerobic ammonium oxidation (anammox) granular sludge reactors were investigated. The performance and physiological characteristics were tracked by continuous-flow monitoring to evaluate the long-term effects. High Cu loading (0.24 g L(-1)d(-1)) exceeded sludge bearing capacity, and precipitation dominated the removal pathway. The Cu distribution migrated from the extracellular polymeric substances-bound to the cell-associated Cu and the Cu forms shifted from the weakly bound to strongly bound fractions over time. Pearson correlation and fluorescence spectra analyses showed that the increase in protein concentrations in the EPS was a clear self-defense response to Cu(II) stress. Two remediation strategies, i.e., ethylenediamine tetraacetic acid (EDTA) washing and ultrasound-enhanced EDTA washing, weakened the equilibrium metal partition coefficient from 5.8 to 0.45 and 0.34 L mg(-1)SS, respectively, thereby accelerating the external diffusion of the Cu that had accumulated in the anammox granules., (Copyright © 2015 Elsevier B.V. All rights reserved.)

While the application of mesophilic anammox process is currently the state of the art, the feasibility of a thermophilic anammox bioprocess is still unclear. In this study, we investigate whether glycine betaine (GB) addition can enhance the thermotolerance of mesophilic anammox biomass in the upflow anaerobic sludge blanket (UASB) reactors fed with synthetic wastewater at a nitrogen loading of approximately 4 kg N m–3 d−1. The results showed that during a long-term operation at 45°C with GB (0, 0.1, 1, 2 mM) addition, anammox performance became worse with the final effluent concentrations of NO2–N of 145 ± 11.6 mg L−1 and nitrogen removal efficiency decreased from 92.3–6.9%. Specific anammox activity decreased from 392.1 ± 12.1–6.0 ± 0.8 mg N g−1 VSS d−1, which were not significantly higher than those in the control reactor. The content of heme c showed a stronger downward trend in T1 (with GB addition) than in the control reactor T0. The qPCR results showed that the relative abundance of Candidatus Kuenenia decreased in both the experimental (from 53.5–28.8%) and control reactors (from 54.1–35.1%). Overall, continuous addition of exogenous GB did not improve the thermotolerance of mesophilic anammox consortia at 45°C. [ABSTRACT FROM AUTHOR]

Anaerobic ammonium oxidation (anammox) is a promising biological technology for treating ammonium-rich wastewaters. However, due to the high sensitivity of anammox bacteria, many external factors have inhibitory effects on this process. As one of the commonly found toxic substances in wastewater, heavy metals (HMs) are possible to cause inhibition on anammox sludge, which then results in a declined treatment performance. Getting insights into the response mechanism of anammox sludge to HMs is meaningful for its application in treating this kind of wastewater. This review summarized the effect of different HMs on treatment performance of anammox bioreactor. In addition, the mechanism of toxication raised by HMs was discussed. Also, the potential mitigation strategies were summarized and the future prospects were outlooked. This review might provide useful information for both scientific research on and engineering application of anammox process for treating HMs containing wastewater. [Display omitted] • The inhibitory effect of heavy metals on anammox was summarized. • The toxic mechanisms of heavy metals and adaptive strategies were analyzed. • The mitigating strategies for heavy metals inhibition were summarized. • The future research prospects were envisaged. [ABSTRACT FROM AUTHOR]