Multi-omics analysis reveals the collaboration and metabolisms of the anammox consortia driven by soluble/non-soluble Fe(III) as the sole iron element.

Iron is recognized as a physiological requirement for anammox bacteria (AnAOB), with Fe(II) considered to be the most effective form. However, Fe(III), instead of Fe(II) is the common iron form in natural and artificial ecosystems. In this study, the nitrogen removal performance and metabolic mechanisms in anammox consortia with soluble and non-soluble Fe(III) as the sole iron element were investigated. After the 150-day operation, the soluble (FeCl3) and insoluble (Fe2O3) Fe(III)-fed anammox systems reached nitrogen removal rates of 71.84 ± 0.80% and 50.20 ± 0.98%, respectively. AnAOB could survive with soluble (FeCl3) or insoluble (Fe2O3) Fe(III) as the sole iron element, reaching relative abundances of 18.49% and 13.16%, respectively. The results show that the formation of anammox core consortia can enable AnAOB's survival to adverse external conditions of Fe(II) deficiency. Metagenomic and metatranscriptomic analysis reveal that Ca. Kuenenia can only uptake Fe(II) into the cell for metabolisms either independently through the extracellular electron transfer or with the cross-feeding of symbiotic microbes. This study provides insight into the utilization and metabolic mechanisms of Fe(III) in Ca. Kuenenia-dominated consortia, and deepens the understanding of anammox core consortia in the nitrogen, carbon, and iron cycling, further promoting the practical applications of anammox processes.

Waste iron scraps promote anammox bacteria to resist inorganic carbon limitation.

The anaerobic ammonium oxidation (anammox) process is adversely affected by the limitation of inorganic carbon (IC). In this research, a new technique was introduced to assist anammox biomass in counteracting the adverse effects of IC limitation by incorporating waste iron scraps (WIS), a cheap and easily accessible byproduct of lathe cutting. Results demonstrated that reducing the influent IC/TN ratio from 0.08-0.09 to 0.04 resulted in a 20 % decrease in the nitrogen removal rate (NRR) for the control reactor, with an average specific anammox activity (SAA) of 0.65 g N/g VSS/day. Nevertheless, the performance of the WIS-assisted anammox reactor remained robust despite the reduction in IC supply. In fact, the NRR and SAA of the WIS-assisted reactor exhibited substantial improvements, reaching approximately 1.86 kg/(m3·day) and 0.98 g N/g VSS/day, respectively. These values surpassed those achieved by the control reactor by approximately 39 % and 51 %, respectively. The microbial analysis confirmed that the WIS addition significantly stimulated the proliferation of anammox bacteria (dominated by Candidatus Kuenenia) under IC limitation. The anammox gene abundances in the WIS-assisted anammox reactor were 3-4 times higher than those in the control reactor. Functional genes prediction based on the KEGG database revealed that the addition of WIS significantly enhanced the relative abundances of genes associated with nitrogen metabolism, IC fixation, and central carbon metabolism. Together, the results suggested that WIS promoted carbon dioxide fixation of anammox species to resist IC limitation. This study provided a promising approach for effectively treating high ammonium-strength wastewater using anammox under IC limitation.

Microbial interactions play a keystone role in rapid anaerobic ammonium oxidation sludge proliferation and biofilm formation.

Two mature anaerobic ammonium oxidation (anammox) consortia with high/low relative abundance of anammox bacteria were inoculated for the rapid sludge proliferation and biofilm formation in this study, named up-flow anaerobic sludge blanket reactor (UASB1) (high) and UASB2 (low), respectively. Results showed that the nitrogen removal efficiency of UASB2 reached 90.94% after the 120-day operation, which was 13% higher than that of UASB1. Moreover, its biomass amounts were 22.18% (biofilm) and 40.96% (flocs) higher than that of UASB1, respectively. Ca. Kuenenia possessed relative abundances of 29.32% (flocs), 27.42% (biofilm) and 31.56% (flocs), 35.20% (biofilm) in the UASB1 and UASB2, respectively. The relative abundances of genes involved in anammox transformation (hzs, nir) and carbon metabolism (fdh, lgA/B/C, acs) were higher in the UASB2, indicating that Ca. Kuenenia might produce acetate and glycogen to enhance microbial interactions. These findings emphasized the importance of microbial interactions in anammox sludge proliferation and biofilm formation.

Succession of microbial communities reveals the inevitability of anammox core in the development of anammox processes.

The lack of anammox seeds is regarded as the bottleneck of anammox-based processes. Although the interactions in anammox consortia have attracted increasing attention, little is known about the influence of inoculated sludge populations on the growth of anammox bacteria. In this study, four sludge of distinct communities mixed with anammox sludge (the relative abundance of Ca. Kuenenia was 1.96 %) were used as the seeds, respectively for the start-up of anammox processes. Notably, all these mixed microbial communities tend to form a similar microbial community, defined as the anammox core, containing anammox-bacteria (22.9 ± 5.9 %), ammonia-oxidizing-bacteria (0.8 ± 0.7 %), nitrite-oxidizing-bacteria (0.2 ± 0.2 %), Chloroflexi-bacteria (0.7 ± 0.4 %), and heterotrophic-denitrification-bacteria (0.3 ± 0.2 %). It also elucidated that the communities of Nitrosomonas-dominated sludge were the closest to the anammox core, and achieved the highest nitrogen-removal rate of 0.73 kg-N m-3 d-1. This study sheds light on the solution to the shortage of anammox seeds in the full-scale wastewater treatment application.

Achieving robust nitritation in a modified continuous-flow reactor: From micro-granule cultivation to nitrite-oxidizing bacteria elimination.

In this study, a modified continuous-flow nitrifying reactor was successfully operated for rapid cultivation of micro-granules and achieving robust nitritation. Results showed that sludge granulation with mean size of ca. 100 µm was achieved within three weeks by gradually increasing settling velocity-based selection pressure from 0.48 to 0.9 m/hr. Though Nitrospira like nitrite-oxidizing bacteria (NOB) were enriched in the micro-granules with a ratio between ammonia-oxidizing bacteria (AOB) and NOB of 5.7%/6.5% on day 21, fast nitritation was achieved within one-week by gradually increasing of influent ammonium concentration (from 50 to 200 mg/L). Maintaining ammonium in-excess was the key for repressing NOB in the micro-granules. Interestingly, when the influent ammonium concentration switched back to 50 mg/L still with the residual ammonium of 15-25 mg/L, the nitrite accumulation efficiency increased from 90% to 98%. Experimental results suggested that the NOB repression was intensified by both oxygen and nitrite unavailability in the inner layers of micro-granules. Unexpectedly, continuous operation with ammonium in excess resulted in overproduction of extracellular polysaccharides and overgrowth of some bacteria (e.g., Nitrosomonas, Arenimonas, and Flavobacterium), which deteriorated the micro-granule stability and drove the micro-granules aggregation into larger ones with irregular morphology. However, efficient nitritation was stably maintained with extremely high ammonium oxidation potential (> 50 mg/g VSS/hr) and nearly complete washout of NOB was obtained. This suggested that smooth and spherical granule was not a prerequisite for achieving NOB wash-out and maintaining effective nitritation in the granular reactor. Overall, the micro-granules exhibited a great practical potential for high-rate nitritation.

The organics-mediated microbial dynamics and mixotrophic metabolisms in anammox consortia under micro-aerobic conditions.

The engineering applications of mainstream anaerobic ammonium oxidation (anammox) have raised increasing attention due to its energy-efficient, however, the organics-mediated microbial dynamics and mixotrophic metabolisms in anammox consortia under micro-aerobic conditions are still elusive. Here, the response of the anammox process to sodium acetate and glucose at a C/N ratio ranging from 0 to 0.5 was investigated under micro-aerobic conditions, respectively. Results showed that the additional glucose could promote the nitrogen removal efficiency (NRE) and nitrogen removal rate (NRR) of anammox processes at a low C/N ratio (0.3), representing 84.00% and 0.53 N kg·m-3·d-1. The introduced organics could regulate the diversity of the microbial community and simplify the microbial relationship in anammox consortia. Anammox could not benefit from the introduced sodium acetate, while glucose could effectively enhance the anammox activity and microbial interactions in anammox consortia. Glucose might also stimulate the mixotrophic mechanism of Ca. Kuenenia, further promotes the proliferation of anammox sludge under micro-aerobic conditions. This study reveals that glucose could positively mediate microbial interactions and mixotrophic metabolism in anammox consortia under micro-aerobic conditions, which raises a new horizon for the proliferation of anammox sludge for mainstream engineering applications.

Microbial dynamics reveal the adaptation strategies of ecological niche in distinct anammox consortia under mainstream conditions.

The feasibility of anammox-based processes for nitrogen-contained wastewater treatment has been verified with different anammox bacteria, however, the ecological niche of anammox bacteria under mainstream conditions is still elusive. In this study, six sludge samples collected from different habitats were utilized to culture anammox bacteria under mainstream conditions, and two distinct anammox genera (Ca. Kuenenia and Ca. Brocadia) with a relative abundance of 6.31% (C1) and 3.09% (C3), respectively, were identified. Notably, the microbial dynamics revealed that anammox bacteria (AMX), ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), Chloroflexi bacteria (CFX), and heterotrophic denitrification bacteria (HDB) were the core members in anammox consortia. However, Ca. Kuenenia and Ca. Brocadia occupied different ecological niches in anammox consortia. The dissolved oxygen and microbial structures of the anammox-continuous stirred tank reactor systems were the main factors to affect their niche differentiation. Meanwhile, comammox might exist in the systems and occupy the ecological niche of AOB in nitrogen cycling. The network analysis suggested that Ignavibacterium could be the associated bacteria in Ca. Kuenenia-dominated consortia, while Ca. Nitrotoga was that in the Ca. Brocadia-dominated consortia. Our findings reveal a valuable reference for the observation of distinct anammox genera under mainstream conditions, which provides theoretical guidance for the engineering application of mainstream anammox-based processes.

A new concept of waste iron recycling for the enhancement of the anammox process.

As a by-product of industry, waste iron scraps (WIS) are low-cost and widely available, which was potential for the development of iron-assisted anammox. In this study, the feasibility of adding WIS to enhance the nitrogen removal of the anammox process (also called WIS-assisted anammox) was demonstrated. Results indicated that the WIS-assisted anammox reactors performed a 15-35% higher nitrogen removal efficiency than that of the control. Compared to the sludge from the control, the sludge from the WIS-assisted anammox reactors had a higher iron content (78-113 g kg-1 SS) and a better specific anammox activity (10.8-15.5 mg N g-1 VSS h-1). The enhanced growth of the anammox bacteria (related to Ca. Kuenenia stuttgartiensis with 99% similarity) in the WIS-assisted anammox reactors was also confirmed by high-throughput sequencing and qPCR. Furthermore, the functional genes predicted by PICRUSt2 revealed a higher level of hydroxylamine oxidoreductase (hao)-like proteins expression of the biomass from the WIS-assisted anammox reactors, implying that the hydroxylamine-related anammox pathway was promoted. Additionally, the observation of cytoplasmic nitrate reductase (narG), copper-containing nitrite reductase (nirK), and nitric oxide reductase (norB) suggested that the introduction of WIS might promote the denitrification ability. This was correlated to the lower ΔNO3-/ΔNH4+ ratio observed in these WIS-assisted anammox reactors. Overall, the WIS-assisted anammox offers a sustainable nitrogen removal process for wastewater treatment with waste iron recycling.

Nitrogen contribution and microbial community of size-fractionated anammox sludge in continuous stirred-tank reactors.

In this study, the microbial diversity of size-fractionated anammox sludge in a well-mixed system and their contribution to nitrogen transformation were investigated. Results showed that small granules (0.2-1.0 mm) contributed to the major part of the nitrogen removal rate (56 %) due to its largest mixed liquor volatile suspended solids (1240 ± 80 mg·L-1). However, large granules (>1.0 mm) possessed the highest relative abundances of Ca. Kuenenia stuttgartiensis and specific anammox activity, representing 49.34 % and 24.45 ± 0.01 mg-N·g-1-mixed liquor volatile suspended solids·h-1, respectively. The microbial diversity decreased as the increase of granular size, resulting in microbial community shifting to a simpler model. Metagenomic analysis showed that fine sludge might be the potential major for NO/N2O production in the mature well-mixed system under inorganic conditions. This study provides guidance for the evaluation of nitrogen contribution by anammox size-fractionated sludge and the inhibition of the potential NO/N2O emission in anammox processes.

Rapid initiation of a single-stage partial nitritation-anammox process treating low-strength ammonia wastewater: Novel insights into biofilm development on porous polyurethane hydrogel carrier.

Media-supported biofilm is a powerful strategy for growth and enrichment of slow-growing microorganisms. In this study, a single-stage nitritation-anammox process treating low-strength wastewater was successfully started to investigate the biofilm development on porous polyurethane hydrogel carrier. Suspended biomass migration into the carrier and being entrapment by its internal interconnected micropores dominated the fast initial colonization stage. Both surface-attached growth and embedded growth of microbes occurred during the following accumulation stage. Fluorescence in situ hybridization analysis of mature biofilm indicated that ammonium-oxidizing bacteria located at the outer layers featured a surface-attached growth, while anammox microcolonies housed in the inner layers proliferated as an embedded-like growth. In this way, the growth rate of anammox bacteria (predominated by Candidatus Kuenenia) could be 0.079 d-1. The anammox potential of the biofilm reactor reached 1.65 ± 0.3 kg/m3/d within two months. This study provides novel insights into nitritation-anammox biofilm formation on the porous polyurethane hydrogel carrier.

Effective utilization of refractory dissolved organic matters in domestic sewage allows to enhanced nitrogen removal by integrated fermentation, nitrification, denitratation and anammox process.

To take full advantage of refractory dissolved organic matters (rDOMs) and generate sufficient nitrate for domestic sewage treatment, this study presented a novel integrated fermentation, nitrification, denitratation and anammox (IFNDA) process in a combined ABR-CSTR reactor. The results showed that an advanced total nitrogen (TN) removal efficiency of 94.1% was obtained after over 190 days operation, resulting in effluent TN concentration as low as 3.6 mg/L. The system nitrogen removal was dominated by anammox with a high proportion of 88.6%. The high conversion rate of acetic acid (54.0%) and volatile fatty acids (64.5%) from rDOMs in domestic sewage by in-situ fermentation drove efficient denitratation. Microflora analysis indicated that the enriched Commamonas (3.5%) and Longilinea (3.3%) dominated hydrolysis and acidogenesis of organics, and Methanosaeta (9.0%) obligated acetoclastic methanogenesis in two-stage fermentation process. Thauera (8.4%) and Candidatus Brocadia (2.5%) were the core bacteria for nitrogen metabolism in the IFNDA system.

Response of nitritation granules to anaerobically pre-treated municipal wastewater at low temperatures in a continuous-flow reactor.

Achieving mainstream nitritation with aerobic granules is attractive based on increasing evidence but generally treating artificial low-ammonium wastewater. Real municipal wastewater is much more complex in composition, the behavior of the nitritation granules would be different when treating real municipal wastewater. Herein, the response of nitritation granules to influent shift from artificial low-ammonium (35-40 mg/L) wastewater to anaerobically pre-treated municipal wastewater (MWWpre-treated) was investigated at low temperatures. Results showed that MWWpre-treated caused the outgrowth of filamentous bacteria on the granule surface and developed into finger-like structures, which in turn resulted in the decrease of the overall granular sludge settleability. Batch-tests and microbial analysis indicated the functional and microbial differentiation between the newly formed fluffy exterior and the original compact granule. The fluffy exterior was dominated by genus Flavobacterium (66.6%) and primarily functioned as COD removal, whereas the nitrifiers (mainly Nitrosomonas) were still located in the compact core and performed nitritation. Moreover, the heterotrophs-dominated fluffy exterior hindered the oxygen transfer towards nitrifiers located in the compact granule and thereby facilitated the stable NOB repression in the granule particularly at low temperatures (<10 °C). Finally, gradual recovery of the granular sludge morphology and settleability occurred after the influent reverted to synthetic low-ammonium wastewater. Overall, this work demonstrated that the feeding of MWWpre-treated only caused morphological changes of the nitritation granules, but its structural and functional stability could be maintained stably.

Characterization of Sodium Alginate-Locust Bean Gum Films Reinforced with Daphnetin Emulsions for the Development of Active Packaging.

In this study, we characterized an active film made of sodium alginate (SA)-locust bean gum (LBG) containing daphnetin-based film. Physicochemical characteristics, as well as antioxidant and antibacterial properties, were investigated. The results showed that the addition of a low concentration of daphnetin increased the flexibility of SA-LBG cling film, leading to an improvement in elongation at break and tensile strength. As the daphnetin content increased, solubility, brightness and transparency of the cling film decreased, and the moisture permeability increased. The antioxidant capacity and antibacterial activity of films with daphnetin were improved compared to those of the basal film. In addition, the cling film formed by adsorption had higher bacterial (Shewanella putrefaciens and Pseudomonas fluorescens) inhibition and antioxidant activity rates than direct film formation. The results indicate that the combination of daphnetin in SA-LBG film provides an active film with antioxidant and antibacterial properties, with potential for the development of food-grade packaging material.

Achieving high-rate partial nitritation with aerobic granular sludge at low temperatures.

Partial nitritation is necessary for the implementation of the mainstream anammox (anaerobic ammonium oxidation) process in wastewater treatment plants. However, the difficulty in outcompeting nitrite-oxidizing bacteria (NOB) at mainstream conditions hinders the performance of partial nitritation. The present work aimed to develop a high-rate partial nitritation process for low-ammonium wastewater treatment at low temperatures by seeding aerobic granules. Experimental results suggested that both stratified structure of nitrifiers developed in the granules and sufficient residual ammonium concentration (18-35 mg N L-1) in the bulk liquid contributed to efficient NOB repression. With the hydraulic retention time progressively shortened from 1.0 to 0.17 h, the influent nitrogen loading rate of the partial nitritation process reached 6.8 ± 0.4 kg N m-3 d-1 even at 10-15 °C. The high concentration (7.5 gVSS L-1) and activity (0.48 g N g-1 VSS d-1 at 11 °C) of granular sludge made the reactor possess an overcapacity evaluated by the ratio between the actual ammonium oxidation rate of the granules and their maximum potential. The overcapacity helped the reactor to face the adverse effect of decreasing temperatures. Overall, this work indicated the great potential of applying aerobic granules to achieve high-rate partial nitritation at mainstream conditions. Moreover, anammox bacteria with a relative abundance of 2.8% was also identified in the partial nitritation granules at the end of this study, suggesting that the granules provided a habitable niche for anammox bacteria growth. Note that these results cannot fully relate to the treatment of real domestic/municipal wastewater, they are a source of important information increasing the knowledge about low temperature partial nitrification.

Hydroxylamine metabolism in mainstream denitrifying ammonium oxidation (DEAMOX) process: Achieving fast start-up and robust operation with bio-augmentation assistance under ambient temperature.

Nitrogen removal from mainstream wastewater via DEnitrifying AMmonium OXidation (DEAMOX) is often challenged by undulated actual temperature and high loading rate. Here, we discovered NH2OH addition (HA) and bio-augmentation (BA) tactics on start-up and operation performance of DEAMOXs (R1 and R2) under ambient temperature (11.3-31.7 °C). Over 340-day operation suggested that R2 received 10 mg/L HA and 1:25 BA ratio (v/v, anammox/partial denitrification sludge) achieved desirable nitrogen removal efficiency (NRE) of 97.22% after 145-day, while R1 under higher BA ratio of 1:12.5 without HA obtained lower NRE (90.86%) after 184-day. Batch tests revealed that nitrate-nitrite transformation ratio reached 98.64% at low COD/NO3--N of 2.6 with HA. Significantly, compared with R2, R1 recovered quickly with satisfactory effluent total nitrogen of 4.21 mg/L despite nitrogen loading rate greater than 0.15 kg N/m3/d and temperature decreased to 14.6 °C. The abundant narG represented high nitrate reduction potential, hzsA and hdh were extensively detected as the symbolisation of anammox metabolism. Thauera, Denitratisoma and unclassified f Comamonadaceae dominated nitrite accumulation. Ca. Brocadia as the dominant anammox bacteria, and its population maintained stable against low temperature and load shocks by NH2OH intensification. Overall, this study offers an opportunity for the wide-applications of DEAMOX treating mainstream wastewater.

Effect of Locust Bean Gum-Sodium Alginate Coatings Combined with High CO2 Modified Atmosphere Packaging on the Quality of Turbot (Scophthalmus maximus) during Refrigerated Storage.

This research was conducted to investigate the effect of active coatings composed of locust bean gum (LBG) and sodium alginate (SA) containing daphnetin emulsions (DEs) combined with modified atmosphere packaging (MAP) on the microbiological and physicochemical properties of turbot during 4 °C refrigerated storage for 32 days. The results revealed that LBG-SA-DE coatings together with high CO2 MAP (60% CO2/35% N2/5% O2) maintained the total viable count (TVC) of H2S-producing bacteria in 4-6 lg CFU/g, which is lower than the limit (7 lg CFU/g). In addition, LBG-SA-DE coatings together with high CO2 MAP (60% CO2/35% N2/5% O2) inhibited the production of odor compounds, including thiobarbituric acid (TBA), trimethylamine-nitrogen (TMA-N), K value, and total volatile basic nitrogen (TVB-N). The low-field NMR analysis (LF-NMR) and magnetic resonance imaging (MRI) indicated that LBG-SA-DE coatings together with high CO2 MAP (60% CO2/35% N2/5% O2) treatments could delay the release of water located in muscle fiber macromolecules or convert it into free water based on muscle fiber destruction, thus maintaining the water content and migration. The results of the sensory evaluation showed that turbot treated with LBG-SA-DE coatings together with MAP could maintain its freshness during refrigerated storage.

Shelf-Life Extension of Refrigerated Turbot (Scophthalmus maximus) by Using Weakly Acidic Electrolyzed Water and Active Coatings Containing Daphnetin Emulsions.

This research was to investigate the effect of weakly acidic electrolytic water (WAEW) treatments combining with the locust bean gum (LBG) and sodium alginate (SA) active coatings, containing daphnetin emulsions on microbiological, physicochemical, and sensory changes of turbot (Scophthalmus maximus) during refrigerated storage at 4°C for 24 days. Results showed that WAEW, together with LBG-SA coatings containing daphnetin emulsions treatments, could significantly lower the total viable count (TVC), H2S-producing bacteria, pseudomonas spp., and psychrotrophic bacteria counts, and inhibit the productions of off-flavor compounds, including the total volatile basic nitrogen (TVB-N), inosine (HxR), and hypoxanthine (Hx). Furthermore, the treatments also prevented textural deterioration, delayed water migration, and had higher organoleptic evaluation results. Therefore, WAEW, together with LBG-SA coatings, containing daphnetin emulsions treatments, had the potential to improve the quality of turbot during refrigerated storage.

[Effects of Cold Acclimation on the Activity of Autotrophic Nitrogen Removal in Granular Sludge and Its Bacterial Population Structure].

Cold acclimation is an effective approach for improving the nitrogen removal performance and operational stability of partial nitritation/ANAMMOX (PN/A) combined processes at low temperatures. To explore the specific effects of cold acclimation on the characteristics of sludge, differentiations in temperature sensitivity, granular morphology, composition of extracellular polymer substance (EPS), and bacterial community structure between PN/A granular sludges cultivated at medium-high temperature (30℃) and acclimated to low temperature (15℃) were investigated in this study. The results of reaction thermodynamics showed that the nitrogen removal performance of the granules acclimated to low temperature (GL) was significantly higher than that of those cultivated at medium-high temperature (GH) under the low temperature (10-20℃), and the apparent activation energy (Ea) of total inorganic nitrogen removal for the former was decreased by 28.4%. Compared with GH, GL had a smaller average particle size of 25.8% and higher EPS contents of 16.6%, resulting in a significant lower settling property. Based on the high-throughput sequencing results, GL exhibited a higher diversity of bacterial community, and a lower relative abundance ratio (0.04) of aerobic ammonium-oxidizing bacteria (Nitrosomonas) and anaerobic ammonium-oxidizing bacteria (Candidatus_Kuenenia) than 0.34 for GH. It indicated that the PN/A granules held a strong ability to retain slow-growing autotrophic bacteria in the system, even under low temperatures. These findings could provide meaningful references for analyzing the self-adaption mechanisms of PN/A sludge to low temperature conditions and promote the industrial application of combined processes.

Enhancing the in-situ enrichment of anammox bacteria in aerobic granules to achieve high-rate CANON at low temperatures.

In this study, a high-rate CANON (Complete Autotrophic Nitrogen-removal Over Nitrite) process was started up successfully by enhancing the in-situ enrichment of anammox bacteria in aerobic granules at conditions relevant for mainstream wastewater treatment. Firstly, to provide nitrite for anammox bacteria growth efficient nitrite-oxidizing bacteria (NOB) repression was rapidly achieved and stably maintained. Both low dissolved oxygen (DO) and ammonium concentrations ratio (DO/NH4+ <0.15) and selective washing-out of NOB-preferred smaller particles at short hydraulic retention time (HRT, 25-15 min) contributed to the NOB repression. Then the stepwise down-regulating DO concentrations from 2.8 to 1.2 mg/L enhanced the enrichment of anammox bacteria in the aerobic granules. The enriched anammox species was dominated by Ca. Brocadia sapporoensis with the estimated growth rate of 0.008-0.013 d-1 at 15 °C. Chloroflexi and Chlorobi-affiliated bacteria were also significantly enriched in the granules, which may benefit the anammox bacteria activity and growth. At the end of this study, the average total nitrogen removal rate and efficiency of the granular CANON process respectively reached 1.26 kg N·m-3·d-1 and 68% treating low-strength ammonium (∼50 mg N·L-1) wastewater under such aggressive conditions (DO = 0.8-1.5 mg/L, HRT< 1.0 h, and T = 15 °C). Overall, the aerobic granules provided a habitable niche for the proliferation and almost complete retention of the anammox bacteria. This study provides a roadmap for in-situ starting up of high-rate CANON process for mainstream wastewater treatment with aerobic granules as inoculum.

Development of a novel denitrifying phosphorus removal and partial denitrification anammox (DPR + PDA) process for advanced nitrogen and phosphorus removal from domestic and nitrate wastewaters.

A novel energy-efficient DPR + PDA (denitrifying phosphorus removal and partial denitrification anammox) process for enhanced nitrogen and phosphorus removal was developed in the combined ABR-CSTR reactor. After 220 days operation, excellent total inorganic nitrogen (TIN) and phosphorus removal (97.57% and 95.66%, respectively) were obtained under external C/NO3--N of 0.7, with the effluent TIN and PO43--P concentrations of 3.51 mg/L and 0.28 mg/L, respectively. At the steady period, DPR contributed major TN removal (58.65%), while PDA mediated an increasingly considerable impact and finally achieved 37.07%, in which anammox accounted for a significant percentage. Batch tests demonstrated that efficient PD with nitrate-to-nitrite transformation ratio of 97.67% supplying stable nitrite for anammox, and phosphorus was mainly removed using nitrate as electron acceptor via DPR with the ideal phosphorus release/uptake rate (7.73/22.17 mgP/gVSS/h). Accumulibacter (6.24%) dominated high phosphorus removal performance, while Thauera (8.26%) and Candidatus Brocadia (2.57%) represented the superior nitrogen removal performance.