The performance of an anaerobic ammonium oxidation (anammox) reactor with the magnetic field of 40 mT was systematically investigated. The total nitrogen removal rate was enhanced by 16% compared with that of the control group. The enhancing mechanism was elucidated from the improved mass transfer efficiency, the complicated symbiotic interspecific relationship and the improved levels of functional genes. The magnetic field promoted formation of the loose anammox granular sludge and the homogeneous and well-connected porous structure to enhance the mass transfer. Consequently, Candidatus Brocadia predominated in the sludge with an increase in abundance of 13%. Network analysis showed that the positive interactions between Candidatus Brocadia and heterotrophic bacteria were strengthened, which established a more complicated stable microbial community. Moreover, the magnetic field increased the levels of hdh by 26% and hzs by 35% to promote the nitrogen metabolic process. These results provided novel insights into the magnetic field-enhanced anammox process.

Currently, in-depth understanding of response characteristics and mechanisms of anammox process under microplastics (MPs) stress remains quite limited. This study investigated the influence of 0.1-1.0 g/L polyethylene terephthalate (PET) on anammox granular sludge (AnGS). Compared with the control, 0.1-0.2 g/L PET did not significantly affect the anammox efficiency, while the anammox activity decreased by 16.2% at 1.0 g/L PET. Integrity coefficient and transmission electron microscopy analysis demonstrated that the strength and structural stability of the AnGS became weaken following exposure to 1.0 g/L PET. With the PET increasing, the abundance of anammox genera and genes related to energy metabolism and cofactors and vitamins metabolism decreased. The reactive oxygen species generated in the interaction between microbial cells and PET resulting in cellular oxidative stress was responsible for inhibiting anammox. These findings give novel insights into the anammox behavior in biological nitrogen removal systems treating PET-loaded nitrogenous wastewater.

Anaerobic ammonium oxidation (anammox) granular sludge is a promising biotechnological process for treating low-carbon nitrogenous wastewater, and is featured with low energy consumption and footprint. Previous theoretical and experimental research on anammox granular sludge processes mainly focused on granulation (flocs → granules), but pay little attention to the granulation cycle including granulation and regeneration. This work reviewed the previous studies from the perspective of anammox granules lifecycle and proposed various sustainable formation mechanisms of anammox granules. By reviewing the anaerobic, aerobic, and anammox granulation mechanisms, we summarize the mechanisms of thermodynamic theory, heterogeneous growth, extracellular polymeric substance (EPS)-based adhesion, quorum sensing (QS)-based regulation, biomineralization-based growth, and stratification of microorganisms to understand anammox granulation. In the regeneration process, the formation of precursors for re-granulation is explained by the mechanisms of physical crushing, quorum quenching and dispersion cue sensing. Based on the granulation cycle mechanism, the rebuilding of the normal regeneration process is considered essential to avoid granule floatation and the wash-out of granules. This comprehensive review indicates that future research on anammox granulation cycle should focus on the effects of filamentous bacteria in denitrification-anammox granulation cycle, the role of QS/ quorum quenching (QQ)-based autoinducers, development of diversified mechanisms to understand the cycle and the cycle mechanisms of stored granules.

The inevitable introduction of biodegradable carbon sources (such as monosaccharides and volatile fatty acids) originating from pretreatment units might affect the performance of the mainstream anaerobic ammonium oxidation (anammox) process. Two model carbon sources (glucose and acetate) were selected to investigate their effects on granule-based anammox systems under mainstream conditions (70 mg total nitrogen (TN) L-1, 15 °C). At a nitrogen loading rate of 2.87 ± 0.80 kg N m-3 d-1, a satisfactory effluent quality (TN < 10 mg L-1) was achieved in the presence of glucose or acetate at a chemical oxygen demand (COD/N) ratio of 0.5. The contribution of anammox to nitrogen removal decreased with increasing COD/N ratio to 1.0 because the expression of anammox functional genes was inhibited, whereas the expression of denitrifying functional genes was promoted. However, the nitrogen removal efficiency of the two considered reactors was maintained above 80 %. Self-stratification of the microbial community along the reactor height facilitated a functional balance through the retention of anammox bacteria in granules but resulted in washout of denitrifying bacteria in flocs under a high-flow pattern. These findings highlighted the advantages of granule-based systems in the mainstream anammox process due to their inherent biomass self-segregation property and the need for the development of targeted biomass retention strategies.

Anaerobic ammonium oxidation (Anammox) granular sludge (AnGS) has poor strength and is prone to disintegration under complex environmental conditions, especially in the presence of complex organic carbon, which renders the Anammox process instable. Herein, with a mixture of landfill leachate and domestic sewage as wastewater, the effect on the properties of AnGS with two small particle size (0.1-0.2 mm) biochars (coconut and peach biochars) addition were investigated at different COD concentrations (150 mg·L-1, 200 mg·L-1, and 250 mg·L-1), as well as at different BOD/TN (B/N) (0.3 and 0.5). Results showed that the nitrogen removal efficiencies decreased from 89 % to 72 % as the COD concentration increased by 100 mg·L-1, while peach biochar reactor had better nitrogen removal performance. Excessive organic carbon supply inhibits AnAOB proliferation and B/N had the most significant effect on AnAOB (p < 0.05). The Polymerase Chain Reaction (PCR) indicated peach biochar reactor get higher activity of anammox-related functional genes (hzsA, hdh).

As a key component of extracellular polymeric substances (EPS), extracellular deoxyribonucleic acid (eDNA) acts as a bridge in maintaining the structural stability of granular sludge. However, its ability of carrying antibiotic resistance genes (ARGs) promotes the gene horizontal transfer, raising a high risk for human health. In this study, a series of batch tests were performed to elucidate the response of anammox granular sludge (AnGS) with different sizes (S-AnGS with diameters lower than 0.9 mm and L-AnGS with diameters of 0.9-2 mm) to the removal of eDNA and corresponding mechanism. The results showed that the highest bioactivity of S-AnGS and L-AnGS was achieved by adding DNase I, and the absolute abundance of hzsA in the systems also increased. The dominant microorganism in each sludge was Candidatus Kuenenia, which maintained a higher relative abundance of 24% in S-AnGS. Settling experiments demonstrated that the permeability of AnGS was positively correlated with the addition of DNase I. The permeability index of granular sludge, Г, rose by 58.54% in S-AnGS and 11.79% in L-AnGS. The absence of eDNA is conducive to the increase in the permeability and porosity of AnGS. Similarity in the functional genes and microbial communities of intracellular and extracellular DNA implied the occurrence of gene transmembrane transfer. The findings enrich our knowledge of eDNA in anammox granules and provide a guidance for the specific control of gene transfer through reducing eDNA.

Nitrogen-removal granular sludge (NRGS) is a promising technology in wastewater treatment, with advantages of efficient nitrogen removal, less footprint, lower sludge production and energy consumption, and is a way for wastewater treatment plants to achieve carbon-neutrality. Aerobic granular sludge (AGS) and anammox granular sludge (AnGS) are two typical NRGS technologies that have attracted extensive attention. Mounting evidence has shown strong associations between NRGS properties and the status of NRGS systems; however, a holistic view is still missing. The aim of this article is to provide an overview of NRGS with an emphasis on characterization. Specifically, the integrated nitrogen transformation pathways inside NRGS and the performance of NRGS treating various wastewaters are discussed. NRGS properties are categorized as physical-, chemical-, biological- and systematical ones, presenting current advances and corresponding characterization technologies. Finally, the future prospects for furthering the mechanistic understanding and engineering application of NRGS are proposed. Overall, the technological advancements in characterization have greatly contributed to understanding NRGS properties, which are potential factors for optimizing the performance and evaluating the working status of NRGS. This review will provide guidance in characterizing NRGS properties and boost the introduction of novel characterization technologies.

The effects of different chemical oxygen demand (COD) concentrations on the anammox granular sludge with Bamboo Charcoal (BC) addition were evaluated in UASB reactor. The results showed that the average total nitrogen (TN) removal efficiency was reduced from 85.9% to 81.4% when COD concentration was increased from 50 to 150 mg/L. However, the TN removal efficiency of BC addition reactors was dramatically 3.1%-6.4% higher than that without BC under different COD concentrations. The average diameter of granular sludge was 0.13 mm higher than that without BC. The settling velocity was increased by elevated COD concentration, while the EPS and VSS/SS were increased with BC addition. The high-throughput Miseq sequencing analyses revealed that the bacterial diversity and richness were decreased under COD addition, and the Planctomycetes related to anammox bacteria were Candidatus Brocadia and Candidatus Kuenenia. The Metagenomic sequencing indicated that the abundance of denitrification related functional genes all increased with elevated COD, while the abundance of anammox related functional genes of decreased. The functional genes related to anammox was hydrazine synthase encoding genes (hzsA, hzsB and hzsB). The average relative abundance of hzs genes in the reactor with BC addition was higher than the control at COD concentrations of 50 mg/L and 150 mg/L. The functional genes of denitrification mediated by BC were higher than those without BC throughout the operation phase. It is interesting to note that BC addition greatly enriched the related functional genes of denitrification and anammox.

Cryopreservation is one of the effective methods for the preservation of anammox granular sludge (AnGS). However, the effects of cooling pretreatment on AnGS are still unclear. In this study, the effects of temperature decrease on AnGS property were investigated by designing different cooling modes: constant at room temperature 20-25 °C (CK), sharp cooling to 4 °C (S4), -20 °C (S20) and stepwise cooling to 4 °C (A4), -20 °C (A20). The results showed that compared with CK, the cooling modes in S4, S20, A4 and A20 improved the physical preservability of AnGS, slowing down the changes of color, shape and structure; and elevated the preservation rate of functional bacteria Planctomycetes (phylum level) and Candidatus Brocadia (genus level). The preservation rate of live cells in different experimental groups was 48.4 ± 1.8%(CK), 61.1 ± 3.3%(S4), 37.8 ± 0.8%(S20), 81.7 ± 4.8%(A4), 61.9 ± 3.1%(A20), respectively. The Anaerobic Ammonium Oxidation Bacteria (AnAOB) in the stepwise cooling mode (A4 and A20) were found to enter the dormant state and form \"dormant zoogloea\", while the AnAOB in the sharp cooling mode (S4 and S20) were observed to enter the shock state with a little change. The findings in this work (especially the dormant state of AnAOB) are helpful to understand the effect of temperature decrease on AnGS and to promote the development of AnGS preservation technology.

Anammox granule is the key support of anammox sludge bed reactor. In this study, the anammox granules from a steady-state reactor were divided into 6 groups to investigate their dimension effects. The results of batch cultivation showed that the anammox granules with VMD (volume surface mean diameter) of 2.17 mm had the maximum SAA (specific anammox activity) of 399.6 ± 37.6 mg-N/(g-VSS·d). The bacterial community analysis demonstrated that Candidatus Kuenenia was the main detectable AnAOB genus in the anammox granules. Q-PCR together with flow cytometry indicated that the total number of viable AnAOB cells ascended with the increasing anammox granular size, suggesting the enhancement of nitrogen removal potential. On the contrary, the mass transfer efficiency descended with the increasing granular size, indicating the restriction of nitrogen removal performance. The maximum SAA was ascribed to the optimal match between nitrogen removal potential and mass transfer efficiency. The results of this study are helpful to comprehend the nitrogen removal capacity of anammox granules and to promote the optimization of anammox process.