Achieving economic and efficient removal of nutrients in mainstream wastewater treatment plants (WWTPs) continues to be a challenging research topic. In this study, a continuous-flow anaerobic/aerobic/anoxic system with sludge double recirculation (AOA-SDR), which integrated partial nitrification (PN), endogenous denitrification (ED) and nitrite-type denitrifying phosphorus removal (nDNPR), was constructed to treat real carbon-limited municipal wastewater. The average effluent concentrations of total inorganic nitrogen (TIN) and PO43--P during the stable operation period were 1.8 and 0.3 mg/L, respectively. PN was achieved with an average nitrite accumulation ratio of 90.4 % by combined strategies. Adequate storage of polyhydroxyalkanoates and glycogen in the anaerobic zone promoted the subsequent nitrogen removal capacity. In the anoxic zone, nitrite served as the main electron acceptor for the denitrifying phosphorus removal process. Mass balance analysis revealed that nDNPR contributed to 23.6 % of TIN removal and 44.7 % of PO43--P removal. The enrichment of Nitrosomonas (0.45 %) and Ellin 6067 (1.31 %), along with the washout of Nitrospira (0.15 %) provided the bacterial basis for the successful implementation of PN. Other dominant endogenous heterotrophic bacteria, such as Dechlormonas (10.81 %) and Candidatus Accumulibacter (2.96 %), ensured simultaneous nitrogen and phosphorus removal performance. The successful validation of integrating PN, ED and nDNPR for advanced nutrient removal in the AOA-SDR process provides a transformative technology for WWTPs.

Reducing CO2 emission and energy consumption is crucial for the sustainable management of wastewater treatment plants (WWTPs). In this study, an algal-bacterial aerobic granular sludge (AGS) system was developed for efficient carbon (C) assimilation and nitrogen (N)/phosphorus (P) removal without the need for mechanical aeration. The photosynthetic O2 production by phototrophic organisms maintained the dissolved oxygen (DO) level at 3-4 mg/L in the bulk liquid, and an LED light control system reduced 10-30% of light energy consumption. Results showed that the biomass assimilated 52% of input dissolved total carbon (DTC), and the produced O2 simultaneously facilitated aerobic nitrification and P uptake with the coexisting phototrophs serving as a C fixer and O2 supplier. This resulted in a stably high total N removal of 81 ± 7% and an N assimilation rate of 7.55 mg/(g-MLVSS∙d) with enhanced microbial assimilation and simultaneous nitrification/denitrification. Good P removal of 92-98% was maintained during the test period at a molar ∆P/∆C ratio of 0.36 ± 0.03 and high P release and uptake rates of 10.84 ± 0.41 and 7.18 ± 0.24 mg/(g- MLVSS∙h), respectively. Photosynthetic O2 was more advantageous for N and P removal than mechanical aeration. This proposed system can contribute to a better design and sustainable operation of WWTPs using algal-bacterial AGS.

In this study, three strains of heterotrophic nitrification-aerobic denitrification (HN-AD) capable of simultaneously removing phosphorus were isolated from activated sludge, and low-temperature coconut shell biochar was prepared. The metabolic effects of combined HN-AD bacteria on the total nitrogen (TN) and total phosphorus (TP) were investigated, and the enhanced efficiency and mechanism of low-temperature biochar on the combined bacteria were also explored. The results indicated that the combined bacteria could adapt to environmental impacts and multiple nitrogen sources. The low-temperature biochar containing more aliphatic carbon and oxygen-containing functional groups enhanced the metabolic activity of combined HN-AD bacteria and accelerated the electron transfer process during nitrogen and phosphorus degradation. The removal efficiencies of TN and TP increased by 68% and 88%, respectively, in the treatment of actual sewage by biochar attached with combined bacteria. The findings form a basis for the engineering utilization of HN-AD and are of great practical significance.

Biological denitrification is still the most important pathway to purifying nitrate-containing wastewater. In this study, pyrite (FeS2 ) and polycaprolactone (PCL) were used as electron donors to construct sole or combined denitrification systems, that is, pyrite-based autotrophic denitrification (PAD) system, PCL-supported heterotrophic denitrification (PHD) system, and split-mixotrophic denitrification system (combined PAD + PHD), all of which were operated under five different hydraulic retention times (HRTs) for 150 days. The results showed that the removal rates (RE) of nitrate (NO3 - -N) and inorganic phosphorus (PO4 3- -P) by PAD were 91% and 94%, respectively, but the effluent sulfate (SO4 2- ) concentration was as high as 168.2 mg/L; the removal rate of NO3 - -N by PHD was higher than 99%, but the PO4 3- -P could not be removed ideally; the removal rates of NO3 - -N and PO4 3- -P by PAD + PHD were higher than 95% and 99%, respectively, and the effluent SO4 2- concentration was only 7.2 mg/L. Through the analysis of the surface scanning electron microscope (SEM) images of the two kinds of media before and after use, it was found that the coupled mode of PAD + PHD was more favorable for biofilm formation than the sole PAD or PHD process, and the microorganisms in the PAD + PHD mode made more full use of electron donors. Moreover, the biomass of the PAD + PHD mode was lower than that of the PAD or PHD process, but the denitrification efficiency of the coupled mode was more efficient, indicating that the functional microorganisms in the PAD + PHD mode might have a certain synergistic effect. PRACTITIONER POINTS: Removal rates of NO3 -, PO4 3 -, and SO4 2 - by PAD were 91%, 94%, and -233%, respectively. Removal rate of NO3 - by PHD exceeded 99%, but PO4 3 - could not be removed ideally. Removal rates of NO3 -, PO4 3 -, and SO4 2 - by PAD + PHD were 95%, 99%, and 86%, respectively. The coupled mode was more favorable for biofilm formation than the sole PAD or PHD. The coupled mode had lower biomass but got more excellent denitrification efficiency.

Acid-pretreated pistachio shells were used as carbon sources to investigate the effects of carbon source dosage on simultaneous nitrogen and phosphorus removal under different carbon/nitrogen (C/N) ratios (7, 9, and 11). Results showed that C/N was positively correlated with mixed liquor suspended solids (MLSS) (R2 = 0.998, p < 0.01) and f value (R2 = 0.975, p < 0.05). Moreover, it was negatively correlated with the sludge volume index (SVI) (R2 =  - 0.959, p < 0.05). C/N was also significantly negatively related to chemical oxygen demand removal rate (R2 =  - 0.986, p < 0.05) and positively related to ammonia nitrogen (NH4+-N), total nitrogen (TN), and total phosphorus (TP) removal rate (p < 0.05), the correlation coefficients were 0.992, 0.990 and 0.994, respectively. In the reactor with C/N of 11, the MLSS concentration and f value were the highest, the SVI was the lowest, and the removal efficiencies of NH4+-N (85.49 % ± 1.96 %), TN (84.19 % ± 1.42 %) and TP (94.10 % ± 1.67 %) were the highest. Furthermore, the relative abundance of denitrifying bacteria was the highest in the reactor. The abundance of nitrifying bacteria and phosphorus-removal bacteria was also relatively high.

Iron-based ecological floating beds (EFBs) are often used to treat the secondary effluent from wastewater treatment plant to enhance the denitrification process. However, the impact and necessity of plants on iron-based EFBs have not been systematically studied. In this research, two iron-based EFBs with and without plants (EFB-P and EFB) were performed to investigate the response of plants on nutrient removal, GHG emissions, microbial communities and functional genes. Results showed the total nitrogen and total phosphorus removal in EFB-P was 45-79% and 48-72%, respectively, while that in EFB was 31-67% and 44-57%. Meanwhile, plants could decrease CH4 emission flux (0-3.89 mg m-2 d-1) and improve CO2 absorption (4704-22321 mg m-2 d-1). Plants could increase the abundance of Nitrosospira to 1.6% which was a kind of nitrifying bacteria dominant in plant rhizosphere. Among all denitrification related genera, Simplicispira (13.08%) and Novosphingobium (6.25%) accounted for the highest proportion of plant rhizosphere and iron scrap, respectively. Anammox bacteria such as Candidatus_Brocadia was more enriched on iron scraps with the highest proportion was 1.21% in EFB-P, and 2.20% in EFB. Principal co-ordinates analysis showed that plants were the critical factor determining microbial community composition. TN removal pathways were mixotrophic denitrification and anammox in EFB-P while TP removal pathways were plant uptake and phosphorus-iron coprecipitation. In general, plants play an important directly or indirectly role in iron-based EFBs systems, which could not only improve nutrients removal, but also minimize the global warming potential and alleviate the greenhouse effect to a certain extent.

This study investigated the interaction between plant biomass and iron scraps and their influence on nitrogen (including nitrate and ammonia) and phosphorus removal in the subsurface flow constructed wetland. The results showed that with the addition of 0.5 g L-1 of plant biomass and 5.0 g L-1 of iron scraps, the nitrate, total nitrogen and total phosphorus removal were simultaneously improved. During 35 days of continuous operation, the plant biomass played main effect on the enhanced denitrification, accounting for about 57%, while iron scraps enhanced the other 43% of nitrogen removal and most phosphorus removal through precipitation inside the wetlands. Iron scraps could benefit the degradation of cellulose into low molecular carbohydrates by 10%, and the biomass could promote the oxidation of iron and increase the total phosphorus removal by 15%. Plant biomass coupled with iron scraps also improved simultaneously the richness, diversity and evenness of microbial community and promoted the abundance of Nitrospira (17.37%) and Thiobacillus (8.46%) in wetlands. In practice, putting iron scraps as matrix and placing plant biomass in the influent region would be a better choice. This research would provide a new method for effective utilization of plant biomass and iron scraps and further treatment of low-polluted wastewater in the wetlands.

The nutrients availability determines efficiency of biological treatment systems, along with the structure and metabolism of microbiota. Herein nutrients deficiencies on aerobic granular sludge were comparatively evaluated, treating wastewater with mass ratios of chemical oxygen demand : nitrogen : phosphorus being 200:20:4, 200:2:4, and 200:20:0.4 (deemed as nutrient-balanced, nitrogen-deficient, and phosphorus-deficient), respectively. Results revealed that both nitrogen and phosphorus deficiencies significantly raised the effluent qualities especially nitrogen removal. However, nitrogen deficiency aroused considerable growth of filamentous bacteria, while granules kept compact structure under phosphorus deficient condition. Extracellular polymeric substances (EPS) also varied in contents and structures in response to different wastewaters. Microbial community structure analysis demonstrated that nitrogen deficiency led to lower richness and higher diversity, while the reverse was observed under phosphorus deficient condition. Nitrogen deficiency mainly induced decrease of nitrifying bacteria, while similarly phosphorus deficiency led to loss of phosphorus accumulating organisms. Dramatic enrichment Candidatus_Competibacter and filamentous Thiothrix were found under nutrients deficiencies, in which the latter explained and indicated filamentous bulking potential especially under nitrogen limited condition. Bacterial metabolism patterns verified the functions of microbial community responding to nutrients via PICRUSt2 prediction mainly by up-regulating cell motility, and cellular processes and signaling. This study could aid understanding of long-term stability of aerobic granular sludge for low-strength wastewater treatment.

To achieve high-efficiency nutrient removal in constructed wetlands (CWs), a novel simultaneous nitrogen and phosphorus removal (SNPR) process was developed by combining nitrification, endogenous denitrification, and denitrifying phosphorus removal. In SNPR process, denitrifying glycogen-accumulating organisms (DGAOs) and denitrifying polyphosphate-accumulating organisms (DPAOs) utilized NOx--N(NO3--N or NO2--N) as electron acceptor and poly-beta-hydroxy-alkanoates (PHAs) as carbon sources for endogenous denitrification and denitrifying phosphorus removal processes. Results from 217 days of operation showed that a high-level of nitrogen removal efficiency of 83.73% was achieved with influent COD/N of 4. The success was attributed to the fact that most of influent carbon sources could be transformed into PHAs before nitrification via enriching DGAOs and DPAOs in CW, which simultaneously improved nitrification and denitrification due to reducing oxygen and carbon sources consumption by aerobic heterotrophs. Phosphorus was mainly removed via denitrifying phosphorus removal, and PO43--P removal efficiency reached up to 87.84% with even common gravel used as substrate. Stoichiometry analysis revealed that DGAOs were the main organisms providing nitrite to DPAOs, suggesting that the effective PO43--P removal under high DGAO abundance condition might be attributed to the coordination of DGAOs and DPAOs in SNRP processes.

The effects of phenol on aerobic granular sludge including extracellular polymeric substances (EPS) and microbial community were investigated for low strength and salinity wastewater treatment. Elevated phenol over 20 mg/L stimulated biological phosphorus removal mainly via co-metabolism with nearly complete phenol degradation, whereas resulted in significant accumulation of nitrate around 4 mg/L. Aerobic granules kept structural stability via enhancing production of extracellular polymeric substances (EPS), especially folds of polysaccharides (PS) and varying functional groups identified through EEM, FTIR and XPS spectral characterizations at increasing phenol loads. Illumina MiSeq sequencing results indicated that elevated phenol decreased the bacterial diversity and richness, and caused remarkable variations in structural and compositions of microbial population. Multiple halophilic bacteria including Stappia, Luteococcus, and Formosa laid the biological basis for stability of aerobic granules and efficient biological nutrients and phenol removal. Redundancy analysis (RDA) suggested the key role of phenol in shaping the relative abundances and predominant genera. This study proved that aerobic granular sludge was feasible for low-saline and phenol-laden low-strength wastewater treatment.