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As one of the most promising autotrophic biological nitrogen removal technology, anaerobic ammonia oxidation (anammox) has gained intense attention for the past decades and several full-scale facilities have been implemented worldwide. However, anammox bacteria are easily affected by disturbed external environmental factors, which commonly leads to the fluctuations in reactor performance. The response of anammox sludge to external stress results in changes in components and structural characteristics of intracellular and extracellular polymer substances. Real-time and convenient spectral analysis of anammox sludge metabolites can give early warning of performance deterioration under external stresses, which is of great significance to the stable operation of bioreactor. This review summarized the research progress on characterizing the intracellular and extracellular metabolites of anammox sludge through spectroscopic techniques. The correlation between anammox sludge activity and its key metabolites was analyzed. Also, the limitations and future prospects of applying spectral analytical techniques for anammox bioreactor monitoring were discussed and outlooked. This review may provide valuable information for both scientific study and engineering application of anammox based nitrogen removal technology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 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 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.)

Anaerobic ammonium oxidation (anammox) is a novel process of deammonification that exhibits superior ecological and economic potential compared to that of traditional heterotrophic processes. Although this process has been successfully implemented in treating high‐strength nitrogen‐contaminated wastewater, it still faces many challenges in treating mainstream municipal wastewater. This review aims to provide an overview of the status and challenges of mainstream anammox‐based processes. The different configurations and crucial factors are discussed in this review. Finally, the future needs for feasible application are stated. Practitioner points: Factors restricting mainstream application of anammox‐based processes are reviewed.Control strategies for selecting and maintaining anammox bacteria are discussed.Recent advances in nitrite production via partial nitrification or denitrification are summarized.Future needs for the feasible application of anammox‐based nitrogen removal technology for mainstream municipal wastewater treatment are outlined. [ABSTRACT FROM AUTHOR]