Highest Total Nitrogen Removal Research Articles

Integration of batch assays and microbial community analysis for partial nitritation and anammox process monitoring

This study investigates the feasibility of using batch assays combined with microbial community analysis to control the partial nitritation/anammox (PN/A) process. A Moving Bed Sequencing Batch Biofilm Reactor (MBSBBR) was operated over 130 days with varying ammonia influent concentrations. The highest anammox activity (22.4mgN/gVSS∙h) was observed at an influent concentration of 200 mg N-NH4/L, correlating with high total inorganic nitrogen (TIN) removal efficiency. However, increasing the ammonia influent concentration to 340mgN/L led to decreased anammox and nitrifying bacteria activities and reduced TIN removal efficiency. Sequencing revealed Candidatus Brocadia as the predominant genus, with its abundance correlating with TIN removal performance. High correlation coefficients (above 0.76) among batch tests highlighted the interdependence of key bacterial activities in the PN/A, emphasizing the need for a balanced microbial community for effective nitrogen removal. Batch assays provided immediate data on metabolic activities, while sequencing offered comprehensive insights into microbial community structure. Integrating these methods allowed for a holistic understanding of the microbial dynamics and their impact on process stability. This approach enhances the accuracy of wastewater treatment monitoring, aiding in the development of effective management strategies for optimizing PN/A processes.

Intensified partial nitrification and microbiome response in a membrane pulsing-aerated biofilm reactor

Membrane aerated biofilm reactor (MABR) has been developed as an energy-saving process for nitrogen removal in municipal sewage treatment, even in high ammonia wastewater. However, the impact of aeration pattern on biofilm microbiome assemblage and nitrification is not well understood. In this study, MABR and membrane pulsing-aerated biofilm reactor (MpABR) were established for partial nitrification under high ammonium loadings. The MABR and MpABR exhibited a 95 % nitrite accumulation rate (NAR), with over 60 % ammonium removal and nitrite conversion achieved after the start-up phase. The MABR and MpABR gradually adapted to the substantial increase in ammonium loading (350 mg NH4+-N·L-1), resulting in stable and high NAR (96 %), ammonium removal efficiency (70 %), and nitrite conversion efficiency (67 %). Orthogonal experiments revealed that ammonium loading rate (ALR) was the primary factor regulating nitrite conversion and oxygen levels. A moderate ALR was preferable to achieve ideal partial nitrification and energy saving. Furthermore, the integrated MpABR-anaerobic ammonium oxidation (Anammox) maintained high total nitrogen (TN) removal (> 90 %) under high ALR conditions. Microbiome analysis revealed that nitrifiers (Nitrosomonas) and denitrifiers (Stenotrophomonas and Arenimonas) alternately dominated the biofilms at different stages. These findings highlight the potential for achieving efficient partial nitrification effects using a MpABR. The excellent TN removal performance of the MpABR-Anammox suggests its wide applicability in energy-efficient wastewater treatment.

Iron-based mixotrophic denitrification for enhancing nitrate removal from municipal secondary effluent: Performance, microbial community dynamics, and economic feasibility

High nitrate content limits the recycling of the secondary effluent of wastewater treatment plants. In the research, one biomass-iron mixture (BIM) filter material based on mixotrophic denitrification mode (heterotrophic and iron-driven autotrophic denitrification) was developed and used to construct a novel denitrification biological filter (BIM-DNBF) for the nitrogen removal of secondary effluent. BIM-DNBF had a short start-up time (approximately 9 days), and high total nitrogen removal (81 %-89 %) without external addition of organic carbon sources during the whole operation. The coexistence of dominant heterotrophic-denitrification-like Pseudomonas and Erysipelothrix as well as iron-driven autotrophic-denitrification-like Citrobacter, Acidovorax, etc. were found in the BIM-DNBF. Moreover, biomass was recognized as one key player in promoting the reduction of Fe3+ to Fe2+, thereby facilitating the occurrence of iron-driven autotrophic denitrification. In addition, BIM-DNBF was assessed to be affordable. These findings provide evidence that BIM-DNBF can be an efficient technology for nitrogen removal of secondary effluent.

Nitrogen removal and nitrous oxide emission from moving bed biofilm reactor (MBBR) under different loads and airflow

Total nitrogen (TN) removal by sewage treatment required the development of nitrifying and denitrifying activities; however, these metabolite activities can produce and emit nitrous oxide (N2O), one of the greenhouse gases (GHG). With the expansion of sewage treatment in the world, compact systems such as moving bed biofilm reactor (MBBR) may be promising, mainly in more populated cities. However, little is known about N2O emission by the MBBR. This work investigated the N-NHx and TN removal, N2O production and emission and nitrogen mass balance in a MBBR under organic load and airflow variation. The highest TN removal (47%) was observed under higher load and higher aeration rate. Meanwhile, the MBBR under lower load and lower airflow had the lowest efficiency of TN and N-NHx removal probably due to a lower nitrifying activity. The N2O emission factors were lower than 1.6% (value estimated by the IPCC for sewage aerobic treatment systems). In addition, in the essay with higher load and lower airflow, half of removed TN was released by settled sludge and there is little research about this subject. Therefore, the MBBR showed to be a promising process for nitrogen removal with little emission of N2O even under different operational conditions that normally can occur in sanitary sewage system in cities around the world.

Performance of a multi-stage hybrid wetland system for the treatment of a dairy effluent

This work evaluates the performance of a pilot-scale hybrid wetland (HW) system for the effluent treatment from a dairy industry, located in Santa Fe (Argentina). The system consists of three stages in a serial configuration: vertical flow (S1) – free water surface (S2) – horizontal subsurface flow (S3) wetlands. The HW was operated during nine months. Wastewater, plant and sediment samples were taken from the system. Main pollutant concentrations were determined in wastewater. N and P concentrations were determined in plant tissues and in sediments to evaluate the accumulation in the system. The mean COD, BOD, TN, and TP removals in the whole HW system were 91 %, 94 %, 76 %, and 64 %, respectively. Organic matter concentration decreased due to aerobic and anaerobic degradation processes in all stages, mainly in S2. S1 presented proper conditions for nitrogen compounds interconversions by ammonification and nitrification processes. The highest total nitrogen removal was achieved in S2. Plant uptake contributed significantly to nutrient retention in S2 and S3. In S2, sediment did not participate significantly in nutrient retention. Phosphorus removal showed the lowest system efficiency. The obtained results determine the system performance under site-specific conditions, which contributes to the scaling process of the treatment wetland.

Spatial separation of nitrifiers and denitrifiers promotes selection and enrichment of polyhydroxyalkanoates storing mixed cultures fed by crude glycerol and propionate wastewater

Polyhydroxyalkanoates (PHA) recovery from industrial wastewater has been highlighted as a promising strategy for a circular bioeconomy. However, the high and varying level of nitrogen in wastewater makes enrichment of mixed microbial culture (MMC) low efficiency. In this study, spatial separation of nitrifiers and denitrifiers was adopted by adding biocarriers in MMC and decreasing the sludge retention time (SRT) to accelerate the enrichment of PHA-storing MMC fed by mixed wastewater containing glycerol and propionate. Nitrifiers and denitrifiers were sustained on biocarriers, obtaining a high total inorganic nitrogen removal and allowing a more efficient selective pressure of a high carbon and nitrogen ratio (C/N) under low SRT conditions. The maximum PHA cell content and relative abundance of PHA-storing bacteria were increased to 60.51 % (SRT 6 d) and 49.62 % (SRT 6 d) with the decrease of SRT, respectively. This study demonstrates an efficient way to highly enrich PHA-storing MMC from crude glycerol, which provide a relevant technical support for high-efficiency enrichment of PHA-storing bacteria in low C/N wastewater.

A novel pure biofilm system based on aerobic denitrification for nitrate wastewater treatment: Exploring the feasibility of high total nitrogen removal under low-carbon condition

To improve the poor denitrification efficiency of traditional biological treatment for low-carbon wastewater, a novel pure biofilm moving bed biofilm reactor (PH-MBBR) based on heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria was proposed. The start-up of PH-MBBR inoculated with HN-AD bacteria and the operating performances and microbial community composition under various C/N ratios were studied. The results showed that at C/N ratio of only 4, the total nitrogen (TN) removal rate of 10.02 mg/(L·h) was achieved under aerobic conditions. Microbial analysis indicates that the synergistic effect of oligotrophic bacteria Thauera and Azoarcus was the main reason for the high pollutant removal at C/N ratio of 4. Furthermore, the analysis of functional genes and enzymes reveals that NorB and NosZ played important roles in nitrate metabolism in PH-MBBR, and aerobic denitrification pathway was NO3–-N → NO → N2O → N2. This study provides a practical and theoretical basis for low-carbon nitrate wastewater treatment.

Microbial Diversity of Biological Filters in Recirculating Aquaculture Systems for Sand Lobster (Panulirus homarus) Rearing

The recirculating aquaculture system (RAS) is a technology that reuses water (often less than 5%) in land-based production for farming aquatic organisms’ water treatment. This technology uses filters, either mechanical or biological, for any aquaculture species rearing. In lobster rearing using RAS, one of the most important factors is the quality of rearing water, which depends primarily on the biochemical activities of the microbial communities in the biofilter. There is a significant correlation between the abundance of nitrifying microbial communities and high total ammonia nitrogen (TAN) removal rate. These organisms can convert ammonia accumulates in the system into nitrate. The application of molecular tools (Metagenome 16S Full-Length Barcoding using Oxford Nanopore Platform) to characterize microbial community diversity has been conducted for this study. The diversity of microbes in RAS biofilter for sand lobster rearing is summarized as follows: Nitrification Bacteria group (β-Proteobacteria, g-Proteobacteria, Nitrospirae); Denitrification/Nitrate Reducing Bacteria (α-Proteobacteria, δ-Proteobacteria, Bacteroidetes/Chlorobi, Firmicutes), Sulfide Oxidation Bacteria (ε-Proteobacteria), Probiotic Bacteria (Bacillus subtilis, Pseudomonas sp., Roseobacter sp.). This study’s results indicate a potential for RAS in sand lobster-intensive aquaculture applications and also discuss future studies’ directions.

Membrane aerated biofilm reactor system driven by pure oxygen for wastewater treatment

Pure oxygen is proposed for wastewater treatment due to its advantages over conventional air aeration. This study investigates a Pure Oxygen-based Membrane Aerated Biofilm Reactor (PO-MABR) for the first time under various operating conditions. The PO-MABR employs a gas-permeable membrane for direct diffusion of low-pressurized pure oxygen to the biofilm, ensuring exceptional carbon and nitrogen removal. The effectiveness of PO-MABR was investigated by varying operational conditions, including temperature, carbon-to-nitrogen ratio, gas pressure, and flow rate. Results indicate superior performance, with a 97% chemical oxygen demand removal and 19% higher total nitrogen removal than Air-Ventilated MABR (A-MABR) due to thicker biofilm and unique microbial structures in PO-MABR. Also, PO-MABR demonstrated resilience to low temperatures and effectively treated both high and low-strength wastewater. The findings emphasize the efficiency of PO-MABR in wastewater treatment, advocating for its adoption due to superior carbon and nitrogen removal across diverse operational conditions.

Horizontal flow aerated constructed wetlands for municipal wastewater treatment: The influence of bed depth

The influence of bed depth on the performance of aerated horizontal constructed wetlands was investigated at the pilot plant scale. Two horizontal flow subsurface constructed wetlands (HF) intensified units of different bed depth (HF1: 0.90 m and HF2: 0.55 m, 0.8 m and 0.5 m water level, respectively) were fitted with forced aeration, while a third one (HFc, 0.55 m bed depth, 0.5 m water level) was used as control and not aerated. The three HF units were operated in parallel, receiving the same municipal wastewater pre-treated in a hydrolytic up-flow sludge blanket anaerobic digester. Applied surface loading rates (SLR) ranged from 20 to 80 g biochemical oxygen demand (BOD5)/m2·d and from 3.7 to 6.7 g total nitrogen (TN)/m2·d, while it ranges from 6 to 23 g BOD5/m2·d and from 1.1 to 1.7 g TN/m2·d in the control unit. Removal of total suspended solids (TSS) and BOD5 was usually close to a 100 % in all units, whilst chemical oxygen demand (COD) removal was higher for the HF1 unit (97 % on average, range of 96–99 %) than for HF2 (92 %, 82–98 %) and HFc (94 %, 86–99 %). TN removal reached on average 33 % (16–43 %) in HFc, 37 % (10–76 %) in HF2 and 51 % (21–79 %) in HF1. High TN removal required a longer aeration time for nitrification and higher effluent recirculation ratio to enhance denitrification. The results indicate that artificial aeration and a high bed depth allows to increase the SLR by a factor of 4 in HF1 but only by a factor of 2 in HF2.

Adaptation of anammox process for nitrogen removal from acidic nitritation effluent in a low pH moving bed biofilm reactor.

Acidic partial nitritation (PN) has emerged to be a promisingly stable process in wastewater treatment, which can simultaneously achieve nitrite accumulation and about half of ammonium reduction. However, directly applying anaerobic ammonium oxidation (anammox) process to treat the acidic PN effluent (pH 4-5) is susceptible to the inhibition of anammox bacteria. Here, this study demonstrated the adaptation of anammox process to acidic pH in a moving bed biofilm reactor (MBBR). By feeding the laboratory-scale MBBR with acidic PN effluent (pH=4.6±0.2), the pH of an anammox reactor was self-sustained in the range of pH 5-6. Yet, a high total nitrogen removal efficiency of over 80% at a practical loading rate of up to 149.7±3.9mg N/L/d was achieved. Comprehensive microbial assessment, including amplicon sequencing, metagenomics, cryosection-FISH, and qPCR, identified that Candidatus Brocadia, close to known neutrophilic members, was the dominant anammox bacteria. Anammox bacteria were found present in the inner layer of thick biofilms but barely present in the surface layer of thick biofilms and in thin biofilms. Results from batch tests also showed that the activity of anammox biofilms could be maintained when subjected to pH 5 at a nitrite concentration of 10mg N/L, whereas the activity was completely inhibited after disturbing the biofilm structure. These results collectively indicate that the anammox bacteria enriched in the present acidic MBBR could not be inherently acid-tolerant. Instead, the achieved stable anammox performance under the acidic condition is likely due to biofilm stratification and protection. This result highlights the biofilm configuration as a useful solution to address nitrogen removal from acidic PN effluent, and also suggests that biofilm may play a critical role in protecting anammox bacteria found in many acidic nature environments.

Pilot Study of Oxic–Anoxic Process under Low Dissolved Oxygen for Nitrogen Removal from Low COD/N Tropical Wastewater

Conventionally, nitrification in biological nitrogen removal (BNR) requires high dissolved oxygen (DO) concentrations (>2 mg L−1), making the process energy intensive. Recent studies have shown that efficient ammonium removal and energy reduction can be realized by operating the nitrification at low DO concentrations (<1 mg L−1). In this study, the low-DO oxic anoxic (low-DO OA) process was operated in a pilot-scale sequencing batch reactor (SBR) over 218 days to evaluate the feasibility of nitrogen removal from low chemical oxygen demand-to-nitrogen ratio (COD/N) tropical municipal wastewater. The results revealed that the low-DO OA process attained high removal efficiency for ammonium (97%) and total nitrogen (TN) (80%) under an average DO concentration of 0.6 mg L−1. The effective TN removal efficiency is attributed to the occurrence of simultaneous nitrification–denitrification (SND) under low DO conditions. Further batch tests revealed that slowly biodegradable COD (sbCOD) in tropical wastewater can support denitrification in the post-anoxic phase, resulting in a high TN removal rate. Compared with high DO concentrations (2 mg L−1), low DO conditions achieved 10% higher TN removal efficiency, with similar ammonium and COD removal efficiency. This study is crucial in promoting the energy efficiency and sustainability of wastewater treatment plants treating low COD/N wastewater.

C/N ratios inform sustainable aerobic denitrification for nitrogen pollution control: Insights into the key parameter from a view of metabolic division

Aerobic denitrification is a feasible and sustainable process for nitrogen removal in environmental protection. The C/N ratio was the vital one that has a significant impact on the performance. However, comprehension of the C/N ratio effect on this bioprocess was not well clarified. In this study, the response of an aerobic denitrifier (Pseudomonas balearica strain RAD-17) to the C/N ratio was investigated from a view of metabolism. It was found that a high total nitrogen (TN) removal efficiency of about 95% was achieved with the dosing C/N ratio of 7.5, while only a slight increase when the dosing C/N ratios were further increased to a high range of 10–30. The carbon validity was out of sync with TN removal and a balance between the two was realizable at a dosing C/N ratio of 7.5 based on the economic consideration. Moreover, weakened assimilation while enhanced respiration along with the increased C/N ratios was revealed, evidenced by the decreased net intracellular nitrogen and increased nitrogenous gas proportions in nitrogen balance analysis. The growth characteristics in terms of cell number and OD600 showed a division for the high flux of nitrate and organics directed to assimilation and respiration, respectively. The expression levels of carbon and nitrate metabolism-related function genes in P. balearica supported the results above. The C/N ratio of 5.0–7.5 was supposed to be the turning region for the metabolism division between assimilation and respiration in aerobic denitrifying. The results are expected to offer a guideline for the decision made for organics supply for the aerobic denitrification process and support its sustainable application.

Step-feed sequence batch reactor filled with surface-modified carriers can enhance nitrogen removal from low C/N ratio wastewater

It is difficult to achieve efficient nitrogen removal from low C/N ratio wastewater if there is no extra carbon source addition. In this study, integrated floating-film activated sludge (IFFAS) process filled with surface-modified carriers (rough, hydrophilic and positive-charged surface) was employed to enhance nitrogen removal performance. By applying step-feed sequence batch reactor strategy, high total nitrogen (TN) removal loading of 0.062 ± 0.010 kg/(m3∙d) could be achieved under the inflow flow ratio of 2:1:1 without external carbon source when treating low C/N ratio wastewater (C/N ratio: 3.5). In comparison, TN removal loading was only 0.031 ± 0.010 kg/(m3∙d) in activated sludge, and 0.057 ± 0.014 kg/(m3∙d) in IFFAS filled with conventional carrier system. The step-feed sequence batch reactor strategy resulted in complete utilization of influent carbon source for denitrification. The low C/N ratio also promoted microbial by-products production that enhanced nitrogen removal. Meanwhile, surface-modified carriers induced the mixed distribution of nitrifiers and denitrifiers. This study proposed a low-cost and efficient approach for nitrogen removal from low C/N ratio wastewater, showing potential in actual application.

A comparative sustainability evaluation of alternative configurations of an urban nitrogen removal solution targeting different pathways

Limiting the introduction of excess nitrogen to natural water sources is a growing priority for water security and environmental health. This poses particular difficulties in urban environments where available land for potential solutions is limited. A promising option is the integrated fixed-film activated sludge (IFAS) process that requires only a small footprint and is capable of high total nitrogen (TN) removal through multiple pathways. In light of the sustainable development goals set out by the United Nations, the present work has sought to compare the sustainability of two TN removal pathways by comparing the technical, economic and environmental performance of their optimum configurations. Through modelling, a single-stage configuration demonstrated the capacity to achieve an effluent TN concentration of 8.7 mg/L by the simultaneous nitrification denitrification pathway when a dissolved oxygen concentration of 3.5 mg/L was provided. Addition of a post-anoxic stage at equal volume to the aerobic stage (1:1 aerobic to anoxic ratio) to target conventional nitrification denitrification could realise an effluent TN concentration of 4.2 mg/L when DO was increased to 4.5 mg/L, although 5.8 mg/L of effluent TN could be achieved with only a 5:1 ratio. In terms of environmental burden and economic costs, analysis of the system's life-cycle under these different configurations indicated considerable asymmetry of the two pathways during the operational phase due mainly to the increased aeration. However in spite of this, the two conventional configurations were ultimately both shown to be more sustainable than that of the simultaneous pathway due to the greater TN removal capacity afforded.

Simultaneous Organic Matter and Nitrogen Removal from Piggery Slaughterhouse Wastewater Using Bioaugmented Gaslift Membrane Bioreactor

AbstractA single bioaugmentation reactor and a side‐stream gaslift membrane bioreactor combined with bioaugmentation are conducted to treat real wastewater from a centralized piggery slaughterhouse in Vietnam. The bioaugmented reactor is inoculated with heterotrophic microorganisms (Bacillus sp.) isolated from piggery slaughterhouse wastewater. The results of a single bioaugmentation experiment show high removal efficiency of chemical oxygen demand (COD) (84.8%–97.5%) and total nitrogen (TN) (69.9%–87.2%) at loading rates of 1.28–3.89 and 0.14–0.37 kg m−3 d−1, respectively. The combined system demonstrates a significantly higher TN removal efficiency (89.0%–96.1%) (p < 0.001), more stable flux (36.0–38.4 L m−2 h−1), and transmembrane pressure (0.95–1.05 bar), and better capacity of separation of solid–liquid phases compared to the single bioaugmentation. High COD and TN removal efficiency is possibly due to assimilation and simultaneous nitrification and denitrification processes. The results of this study also indicate the feasibility and propitious efficiency of the bioaugmented gaslift membrane bioreactor for piggery slaughterhouse wastewater treatment.

Effects of scouring on membrane aerated biofilm reactor performance and microbial community composition

This study investigated the effects of scouring on Membrane Aerated Biofilm Reactors (MABRs). Laboratory-scale MABRs were operated under conditions typical for municipal wastewater. Scouring was induced by bubbling dinitrogen gas through the reactor bulk at low and high frequencies (LF and HF).At low nitrogen surface loads, almost complete ammonium removal was observable while HF scouring resulted in less total nitrogen (TN) removal compared to LF scouring. High nitrogen surface loads combined with HF scouring resulted in a higher TN removal as LF scouring.HF scouring resulted in around four times more sludge production and less residual biofilm mass compared to LF scouring. 16S amplicon sequencing of the biofilm, detached biomass and flocs revealed major differences between the microbial community compositions of these fractions. These results indicate that by varying the scouring strategy is a potential control mechanism for MABR operation and can help to reach specific treatment targets.

Effects of combined antibiotics on nitrification, bacteria and antibiotic resistance genes in activated sludge: Insights from legacy effect of antibiotics

The effect of combined antibiotics exposure on nitrogen removal, microbial community assembly and proliferation of antibiotics resistance genes (ARGs) is a hotspot in activated sludge system. However, it is unclear that how the historical antibiotic stress affects the subsequent responses of microbes and ARGs to combined antibiotics. In this study, the effects of combined sulfamethoxazole (SMX) and trimethoprim (TMP) pollution on activated sludge under legacy of SMX or TMP stress with different doses (0.005-30 mg/L) were investigated to clarify antibiotic legacy effects. Nitrification activity was inhibited under higher level of combined exposure but a high total nitrogen removal (∼70%) occurred. Based on the full-scale classification, the legacy effect of past antibiotic stress had a marked effect on community composition of conditionally abundant taxa (CAT) and conditionally rare or abundant taxa (CRAT). Rare taxa (RT) were the keystone taxa in the microbial network, and the responses of hub genera were also affected by the legacy of antibiotic stress. Nitrifying bacteria and genes were inhibited by the antibiotics and aerobic denitrifying bacteria (Pseudomonas, Thaurea and Hydrogenophaga) were enriched under legacy of high dose, as were the key denitrifying genes (napA, nirK and norB). Furthermore, the occurrences and co-selection relationship of 94 ARGs were affected by legacy effect. While, some shared hosts (eg., Citrobacter) and hub ARGs (eg., mdtD, mdtE and acrD) were identified. Overall, antibiotic legacy could affect responses of activated sludge to combined antibiotic and the legacy effect was stronger at higher exposure levels.

Study on the effect of ratio of microalgae Chlorella sp. and activated sludge to remove nutrients and organic matter for low C/N wastewater

Microalgae and activated sludge were co-cultured in photobioreactor (PBR) systems with different ratios (1:0, 3:1, 1:1, 0:1 wt/wt) to determine an optimal ratio for organic matter and nutrient removal. The results showed that the PBR systems at 1:0 and 3:1 ratios have higher total nitrogen (TN) removal rates than at two others. The highest TN removal at the ratio of 1:0 achieved 96% and at the ratio 3:1 achieved 90% after six days of operation. In addition, the single-microalgae culture and co-culture microalgae-activated sludge systems had higher TP removal than that in the single-activated sludge one. The highest TP removal of 98.8% was achieved at 1:0 ratio after nine days. The 3:1 and 1:1 ratios had significantly higher COD removal rates than the other ratios with 131 mg/l/d and 118 mg/l/d, respectively. After four days of operation, the 3:1 ratio reached the highest specific removal rate of 132.7 mg/l/d. This study showed that the 3:1 ratio of microalgae co-culture and activated sludge is an optimal ratio for wastewater treatment-based microalgae application.

Successful Application of Anammox Using the Hybrid Autotrophic-Heterotrophic Denitrification Process for Low-Strength Wastewater Treatment.

Directly integrating anammox into sewage treatment is attractive, but anammox bacteria (AnAOB) enrichment is complex due to vicious competition from heterotrophic bacteria (HB). A novel strategy of optimal organics management using a preanaerobic stage and subsequent limited-oxygen conditions (0.32 ± 0.15 mg-O2/L) is applied, and a hybrid autotrophic-heterotrophic denitrification process is developed to treat sewage-like wastewater with a COD/N ratio of 3.1 for 420 days. The stable process was achieved, and a high total nitrogen removal rate of 0.53 kg-N/(m3·d) was obtained compared to conventional nitrification/denitrification. The 16S rRNA high-throughput sequencing analysis suggested that the relative abundance of the nonendogenous HB (Denitratisoma and Thauera) was drastically reduced (P ≤ 0.001), whereas the endogenous denitrifying HB (Candidatus (Ca.) Competibacter) was significantly enriched in the anammox granules (9.98%, P ≤ 0.001). Moreover, Ca. Competibacter as an inner core and Nitrospira and Ca. Brocadia as an outside coating of the anammox granules indicated the cooperation of AnAOB with HB as revealed by laser-scanning confocal microscopy and qPCR. In situ tests further confirmed nitrite from two pathways (partial nitritation and endogenous partial denitritation) that favored AnAOB enrichment. Optimal organics management can mitigate the competition of HB with AnAOB by redirecting the metabolic pathways and microbial community, which is critical to directly integrating anammox into sewage treatment.