Phytoremediation performance of mixed aquatic plantings and their rhizosphere microbial community in pilot-scale constructed wetlands.

Phytoremediation is a low-cost, environmentally friendly, and sustainable technology that can utilize vegetation and microorganisms to avoid eutrophication and purifying water environment. The ability of five different living aquatic plants of nitrogen (N), phosphorus (P), and chemical oxygen demand (CODcr) removal were investigated in pilot scale constructed wetlands (CWs). Aquatic plant mixes significantly improved CODcr removal and plant tissue uptake of nitrogen and phosphorus. The wetland performance of mixed plantings was also influenced by the specific species. When assessing pollutant removal in CWs, PNM performed better within mixtures, a possible synergistic effect, while TNV performed poorly, a possible antagonist effect. The nutrient uptake within plant tissues by mixed plants was always a synergistic effect. Aquatic plant mixes significantly increased the rhizosphere microbial diversity and promoted the growth of functional heterotrophic denitrifying flora.

A bio-inspired co-simulation crawling robot enabled by a carbon dot-doped dielectric elastomer.

Flexible actuation materials play a crucial role in biomimetic robots. Seeking methods to enhance actuation and functionality is one of the directions in which actuators strive to meet the high-performance and diverse requirements of environmental conditions. Herein, by utilizing the method of adsorbing N-doped carbon dots (NCDs) onto SiO2 to form clusters of functional particles, a NCDs@SiO2/PDMS elastomer was prepared and its combined optical and electrical co-stimulation properties were effectively harnessed to develop a biomimetic crawling robot resembling Rhagophthalmus (firefly). The introduction of NCDs@SiO2 cluster particles not only effectively improves the mechanical and dielectric properties of the elastomer but also exhibits fluorescence response and actuation response under the co-stimulation of UV and electricity, respectively. Additionally, a hybrid dielectric elastomer actuator (DEA) with a transparent SWCNT mesh electrode exhibits two notable advancements: an 826% increase in out-of-plane displacement under low electric field stimulation compared to the pure matrix and the ability of NCDs to maintain a stable excited state within the polymer for an extended duration under UV-excitation. Simultaneously, the transparent biomimetic crawling robot can stealthily move in specific environments and fluoresce under UV light.

High-Strength Smart Microneedles with "Offensive and Defensive" Effects for Intervertebral Disc Repair.

Intervertebral disc degeneration (IVDD) is a global public health issue. The injury of annulus fibrosus (AF) caused by acupuncture or discectomy can trigger IVDD again. However, there is currently no suitable method for treating AF injury. In this study, the high-strength smart microneedles (MNs) which can penetrate the AF tissue through a local and minimally invasive method, and achieve remote control of speeded-up release of the drug and hyperthermia by the Near Infrared is developed. The PDA/GelMA composite MNs loaded with diclofenac sodium are designed to extracellularly "offend" the inflammatory microenvironment and mitigate damage to cells, and intracellularly increase the level of cytoprotective heat shock proteins to enhance the defense against the hostile microenvironment, achieving "offensive and defensive" effects. In vitro experiments demonstrate that the synergistic treatment of photothermal therapy and anti-inflammation effectively reduces inflammation, inhibits cell apoptosis, and promotes the synthesis of the extracellular matrix (ECM). In vivo experiments show that the MNs mitigate the inflammatory response, promote ECM deposition, reduce the level of apoptosis, and restore the biomechanical properties of the intervertebral disc (IVD) in rats. Overall, this high-strength smart MNs display promising "offensive and defensive" effects that can provide a new strategy for IVD repair.

Copper-Zinc-Doped Bilayer Bioactive Glasses Loaded Hydrogel with Spatiotemporal Immunomodulation Supports MRSA-Infected Wound Healing.

Developing biomaterials with antimicrobial and wound-healing activities for the treatment of wound infections remains challenging. Macrophages play non-negligible roles in healing infection-related wounds. In this study, a new sequential immunomodulatory approach is proposed to promote effective and rapid wound healing using a novel hybrid hydrogel dressing based on the immune characteristics of bacteria-associated wounds. The hydrogel dressing substrate is derived from a porcine dermal extracellular matrix (PADM) and loaded with a new class of bioactive glass nanoparticles (BGns) doped with copper (Cu) and zinc (Zn) ions (Cu-Zn BGns). This hybrid hydrogel demonstrates a controlled release of Cu2+ and Zn2+ and sequentially regulates the phenotypic transition of macrophages from M1 to M2 by alternately activating nucleotide-binding oligomerization domain (NOD) and inhibiting mitogen-activated protein kinases (MAPK) signaling pathways. Additionally, its dual-temporal bidirectional immunomodulatory function facilitates enhanced antibacterial activity and wound healing. Hence, this novel hydrogel is capable of safely and efficiently accelerating wound healing during infections. As such, the design strategy provides a new direction for exploring novel immunomodulatory biomaterials to address current clinical challenges related to the treatment of wound infections.

Distinctive species interaction patterns under high nitrite stress shape inefficient denitrifying phosphorus removal performance.

Denitrifying phosphorus removal using nitrite as an electron acceptor is an innovative, resource-efficient approach for nitrogen and phosphorus removal. However, the inhibitory effects of nitrite on anoxic phosphorus uptake and process stability are unclear. This study investigated the total phosphorus removal performance under nitrite stress and analyzed microbiome responses in 186 sludge samples. The results indicated that the total phosphorus removal rates and dominant taxon abundance were highly similar under nitrite stress. High nitrite stress induced a community-state shift, leading to unstable dynamics and decreased total phosphorus removal. This shift resulted from increased species cooperation. Notably, the shared genera OLB8 and Zoogloea under non-inhibitory nitrite stress, suggesting their vital roles in mitigating nitrite stress by enhancing carbon and energy metabolism. The response patterns of these bacterial communities to high nitrite stress can guide the design and optimization of high-nitrogen wastewater reactors.

Can network attention effectively stimulate corporate ESG practices?-Evidence from China.

Environmental, social, and governance (ESG) has emerged as a widespread concern for all societal segments. This study aims to explore the influence of network attention on corporate ESG practices from an investor perspective. We find that rising network attention significantly increases corporate ESG practices. Specifically, network attention plays the role of external monitoring, image promotion incentives, and mitigation of financing constraints to make companies willing to challenge ESG practices. Additionally, the promoting effect of network attention on firms' ESG practices was more significant in higher marketization processes, severely competitive industries, and non-state enterprises. In the internet era, companies must pay attention to the flow effect caused by network attention, meet stakeholder demands, and pursue long-term sustainable development.

Thickness-Dependent Room-Temperature Optoelectronic Gas Sensing Performances of 2D Nonlayered Indium Oxide Crystals from a Liquid Metal Printing Process.

Due to excellent gas sensing performances, such as high responsivity, good selectivity, and long-term stability, two-dimensional (2D) nonlayered metal oxide semiconductors have attracted wide attention. However, their thickness-dependent gas sensing behaviors are rarely investigated, which is critical in the development of practical 2D sensors. In this work, 2D In2O3 crystals with a range of thicknesses are realized by extracting the self-limited oxide layer from the liquid indium droplets in a controlled environment. A strong thickness-dependent optoelectronic NO2 sensing behavior at room temperature is observed. While full reversibility and excellent selectivity toward NO2 are shown despite the thicknesses of 2D In2O3, the 1.9 nm thick In2O3 exhibits a maximum response amplitude (ΔI/Ig = 1300) for 10 ppm of NO2 at room temperature with 365 nm light irradiation, which is about 18, 58, and 810 times larger than those of its 3.1 nm thick, 4.5 nm thick, and 6.2 nm thick counterparts, respectively. The shortest response and recovery times (i.e., 40 s/48 s) are demonstrated for the 1.88 nm thick In2O3 as well. We correlate such a phenomenon with the change in the In2O3 band structure, which is influenced by the thickness of 2D crystals. This work provides in-depth knowledge of the thickness-dependent gas-sensing performances of emerging 2D nonlayered metal oxide crystals, as well as the opportunities to develop next-generation high-performing room-temperature gas sensors.

Luteolin-loaded biocomposites containing tantalum and polyimide with antibacterial effects for facilitating osteogenic differentiation and bone bonding.

Polymer-based composites are considered promising candidates for bone repair as they possess some outstanding advantages over ceramic/metallic/polymeric biomaterials. Tantalum (Ta)/polyimide (PI) biocomposites (PT) containing 20 v% (PT20) and 40 v% (PT40) Ta nanoparticles were fabricated, and luteolin (LU) was loaded on PT40 (LUPT40). Compared with PT20 and PI, PT40 with a high Ta content displayed high surface behaviors (e.g., roughness, surface energy, and hydrophilicity). PT40 remarkably improved cell adhesion and multiplication, and LUPT40 with LU displayed further enhancement in vitro. Moreover, LUPT40 evidently boosted osteoblastic differentiation while suppressing osteoclastic differentiation. Furthermore, LUPT40 exhibited good antibacterial effects because of the slow release of LU. The in vivo results confirmed that PT40 markedly promoted bone formation and LUPT40 further enhanced bone formation/bone bonding. In brief, the incorporation of Ta particles improved the surface behaviors of PT40, which stimulated cell response/bone formation. Moreover, the slow release of LU from LUPT40 not only promoted cell response/bone formation but also enhanced bone bonding. The synergistic effects of Ta and LU release from LUPT40 enhanced bone formation/bone bonding. Therefore, LUPT40 would have great potential for the repair of bear-loading bone.

Using Nvivo to Analyze the Impact of Computer Simulation of Parent-Child Cooperative Art Activities on the Growth of Preschool Children.

Psychology originally defined parent-child conflict in terms of interpersonal relationships, where parent-child conflict is a process of inconsistent attitudes between parents and children that occurs in a family setting. For this end, we aims to investigate the influence of parental awareness on preschoolers' perception of parent-child conflict and the mediating role of preschoolers' self-esteem. This paper proposes a dynamic parent-child relationship discovery algorithm based on the impact of parent-child cooperation activities on preschool children's development. We applied SPSS and Mplus statistical software for data processing and analysis, and Nvivo 11.0 qualitative software for validation and analysis. The reliability of preschool children's perceived parent-child conflict and sub-dimensions were: 0.901, 0.799, 0.791, 0.811, 0.729; the total scale and the retest reliability of each dimension were: 0.914, 0.837, 0.836, 0.792, 0.711. Validated factor analysis using Mplus: RMSEA = 0.075, TLI = 0.856, CFI = 0.876, SRMR = 0.064.

Revealing the infiltration process and retention mechanisms of surface applied free DNA tracer through soil under flood irrigation.

It has been recently demonstrated that free DNA tracers have the potential in tracing water flow and contaminant transport through the vadose zone. However, whether the free DNA tracer can be used in flood irrigation area to track water flow and solute/contaminant transport is still unclear. To reveal the infiltration process and retention mechanisms of surface applied free DNA tracer through soil under flood irrigation, we tested the fate and transport behavior of surface applied free DNA tracers through packed saturated sandy soil columns with a 10 cm water head mimicking flood irrigation. From the experimental breakthrough curves and by fitting a two-site kinetic sorption model (R2 = 0.83-0.91 and NSE = 0.79-0.89), adsorption/desorption rates could be obtained and tracer retention profiles could be simulated. Together these results revealed that 1) the adsorption of free DNA was dominantly to clay particles in the soil, which took up 1.96 % by volume, but took up >97.5 % by surface area and densely cover the surface of sand particles; and 2) at a pore water pH of 8.0, excluding the 4.9 % passing through and 3.1 % degradation amount, the main retention mechanisms in the experimental soil were ligand exchange (42.0 %), Van der Waals interactions (mainly hydrogen bonds), electrostatic forces and straining (together 44.7 %), and cation bridge (5.3 %). To our knowledge, this study is the first to quantify the contribution of each of the main retention mechanisms of free synthetic DNA tracers passing through soil. Our findings could facilitate the application of free DNA tracer to trace vadose zone water flow and solute/contaminant transport under flood irrigation and other infiltration conditions.

Quercetin-loaded porous biocomposite of polyimide and molybdenum disulfide nanosheets with antibacterial capability for boosting osteoblastic differentiation and bone-bonding.

Implant instability and bacterial infection are the two main reasons for the failure of bone implantation. Herein, a porous biocomposite containing polyimide (PI) and 40 w% molybdenum disulfide (MoS2) nanosheets (PM40) was fabricated, and quercetin (QT) was loaded onto the porous surface of PM40 (PMQT). Incorporation of MoS2 nanosheets into PI remarkably increased the compressive strength, water absorption and protein absorption of PM40. PM40 exhibited good antibacterial capability owing to presence of MoS2, while PMQT displayed the further enhancement of antibacterial capability because of loading of QT. PM40 with MoS2 significantly stimulated the osteoblastic differentiation of bone mesenchymal stem cells in vitro, and PMQT with QT displayed further enhancement. In comparison with PI and PM40, PMQT significantly inhibited the osteoclastic differentiation thanks to the sustained-release of QT that suppressed the formation of osteoclasts and expression of osteoclastic genes. Moreover, PM40 with MoS2 accelerated osteogenesis and bone-bonding in vivo, and PMQT with QT displayed further enhancement. In summary, the cooperative effect of MoS2 and QT significantly improved osteoblastic differentiation and ameliorated bone-bonding in vivo. Accordingly, PMQT displayed marvelous osteogenic and antibacterial effects, which would have the potential for repair of load-bearing bone.

Engineered biomimetic micro/nano-materials for tissue regeneration.

The incidence of tissue and organ damage caused by various diseases is increasing worldwide. Tissue engineering is a promising strategy of tackling this problem because of its potential to regenerate or replace damaged tissues and organs. The biochemical and biophysical cues of biomaterials can stimulate and induce biological activities such as cell adhesion, proliferation and differentiation, and ultimately achieve tissue repair and regeneration. Micro/nano materials are a special type of biomaterial that can mimic the microstructure of tissues on a microscopic scale due to its precise construction, further providing scaffolds with specific three-dimensional structures to guide the activities of cells. The study and application of biomimetic micro/nano-materials have greatly promoted the development of tissue engineering. This review aims to provide an overview of the different types of micro/nanomaterials, their preparation methods and their application in tissue regeneration.

Reclaimed water influences bacterioplankton and bacteriobenthos communities differently in river networks.

Reclaimed water reuse is a promising strategy for addressing water scarcity; however, its potential ecological impact remains largely unknown. In particular, the differential effects of reclaimed water on microbial communities in various habitats remain poorly understood. Here, we aimed to elucidate the distinct effects of reclaimed water on bacterioplankton and bacteriobenthos communities in reclaimed water-receiving river networks from multiple perspectives, including community structure, co-occurrence patterns, assembly mechanisms, and nitrogen cycle function. Significant differences in microbial composition were observed between the plankton and benthic habitats, and the average numbers of amplicon sequence variants (ASVs) that originated from the wastewater treatment plants (WWTP) sites were 310.0 and 613.3, respectively, indicating a stronger association between WWTP and benthic habitats. Random forest and network co-occurrence analyses identified the genus Clostridium_sensu_stricto as a biomarker and key module hub. The assembly of bacteriobenthos communities was driven primarily by deterministic processes (58.74% for River-S and 58.94% for WWTP-S), whereas for bacterioplankton communities, this proportion was reduced to 18.02% (River-W) and 19.09% (WWTP-W). The qPCR revealed a large difference in abundance between the N cycling related genes of bacteriobenthos (average 2.47 × 106 copies/ng) and bacterioplankton (average 3.11 × 103 copies/ng) communities, and different interaction patterns with functional genes. Variance partitioning analysis (VPA) indicated that nitrogen was the most important pollutant, affecting the structure and ecological functions of microbial communities. Moreover, pathway analysis suggested that the reuse of reclaimed water may have enhanced the N-cycling functions of microbial communities and the emission of nitrous oxide.

Biodegradable composites of poly(propylene carbonate) mixed with silicon nitride for osteogenic activity of adipose-derived stem cells and repair of bone defects.

Absorbable polymers have attracted increasing attention in the field of bone regeneration in recent years for their degradation. Compared with other degradable polymers, polypropylene carbonate (PPC) has several advantages such as biodegradation and relatively cheap raw materials. Most importantly, PPC can degrade into water and carbon dioxide totally which does not give rise to local inflammation and bone resorption in vivo. However, pure PPC has not presented excellent osteoinductivity properties. In order to enhance the osteoinductivity of PPC, silicon nitride (SiN) was employed due to its excellent mechanical properties, biocompatibility and osteogenesis compared with the other common materials such as hydroxyapatite and calcium phosphate ceramics. In this study, composites of PPC mixed with different contents of SiN were prepared successfully (PSN10 with 10 wt% SiN content, and PSN20 with 20 wt% SiN content). The characterization of the composites suggested that PPC mixed with SiN evenly and PSN composites presented stable properties. The results in vitro revealed that the PSN20 composite possessed satisfactory biocompatibility and exerted better osteogenic differentiation effects on adipose-derived stem cells (ADSCs). In particular, the PSN20 composite accelerated the healing of bone defects better and degraded with the process of bone healing in vivo. Overall, the PSN20 composite exhibited better biocompatibility, induced osteogenic differentiation of ADSCs and promoted healing of bone defects, due to which the PSN composite is considered as a potential candidate for treating bone defects in the field of bone tissue engineering.

Building Osteogenic Microenvironments with a Double-Network Composite Hydrogel for Bone Repair.

The critical factor determining the in vivo effect of bone repair materials is the microenvironment, which greatly depends on their abilities to promote vascularization and bone formation. However, implant materials are far from ideal candidates for guiding bone regeneration due to their deficient angiogenic and osteogenic microenvironments. Herein, a double-network composite hydrogel combining vascular endothelial growth factor (VEGF)-mimetic peptide with hydroxyapatite (HA) precursor was developed to build an osteogenic microenvironment for bone repair. The hydrogel was prepared by mixing acrylated ß-cyclodextrins and octacalcium phosphate (OCP), an HA precursor, with gelatin solution, followed by ultraviolet photo-crosslinking. To improve the angiogenic potential of the hydrogel, QK, a VEGF-mimicking peptide, was loaded in acrylated ß-cyclodextrins. The QK-loaded hydrogel promoted tube formation of human umbilical vein endothelial cells and upregulated the expression of angiogenesis-related genes, such as Flt1, Kdr, and VEGF, in bone marrow mesenchymal stem cells. Moreover, QK could recruit bone marrow mesenchymal stem cells. Furthermore, OCP in the composite hydrogel could be transformed into HA and release calcium ions facilitating bone regeneration. The double-network composite hydrogel integrated QK and OCP showed obvious osteoinductive activity. The results of animal experiments showed that the composite hydrogel enhanced bone regeneration in skull defects of rats, due to perfect synergistic effects of QK and OCP on vascularized bone regeneration. In summary, improving the angiogenic and osteogenic microenvironments by our double-network composite hydrogel shows promising prospects for bone repair.

Nitrite-resistance mechanisms on wastewater treatment in denitrifying phosphorus removal process revealed by machine learning, co-occurrence, and metagenomics analysis.

Nitrite is a key intermediate in nitrogen metabolism that determines microbial transformations of N and P, greenhouse gas (N2O) emissions, and system nutrient removal efficiency. However, nitrite also exerts toxic effects on microorganisms. A lack of understanding of high nitrite-resistance mechanisms at community- and genome-scale resolutions hinders the optimization for robustness of wastewater treatment systems. Here, we established nitrite-dependent denitrifying and phosphorus removal (DPR) systems under a gradient concentration of nitrite (0, 5, 10, 15, 20, and 25 mg N/L), relying on 16S rRNA gene amplicon and metagenomics to explore high nitrite-resistance mechanism. The results demonstrated that specific taxa were adopted to change the metabolic relationship of the community through phenotypic evolution to resist toxic nitrite contributing to the enhancement of denitrification and inhibition of nitrification and phosphorus removal. The key specific species, Thauera enhanced denitrification, whereas Candidatus Nitrotoga decreased in abundance to maintain partial nitrification. The extinction of Candidatus Nitrotoga induced a simpler restructuring-community, forcing high nitrite-stimulating microbiome to establish a more focused denitrification rather than nitrification or P metabolism in response to nitrite toxicity. Our work provides insights for understanding microbiome adaptation to toxic nitrite and giving theoretical support for operation strategy of nitrite-based wastewater treatment technology.

The transcribed centromeric gene OsMRPL15 is essential for pollen development in rice.

Centromeres consist of highly repetitive sequences that are challenging to map, clone, and sequence. Active genes exist in centromeric regions, but their biological functions are difficult to explore owing to extreme suppression of recombination in these regions. In this study, we used the CRISPR/Cas9 system to knock out the transcribed gene Mitochondrial Ribosomal Protein L15 (OsMRPL15), located in the centromeric region of rice (Oryza sativa) chromosome 8, resulting in gametophyte sterility. Osmrpl15 pollen was completely sterile, with abnormalities appearing at the tricellular stage including the absence of starch granules and disrupted mitochondrial structure. Loss of OsMRPL15 caused abnormal accumulation of mitoribosomal proteins and large subunit rRNA in pollen mitochondria. Moreover, the biosynthesis of several proteins in mitochondria was defective, and expression of mitochondrial genes was upregulated at the mRNA level. Osmrpl15 pollen contained smaller amounts of intermediates related to starch metabolism than wild-type pollen, while biosynthesis of several amino acids was upregulated, possibly to compensate for defective mitochondrial protein biosynthesis and initiate consumption of carbohydrates necessary for starch biosynthesis. These results provide further insight into how defects in mitoribosome development cause gametophyte male sterility.

Base type determines the effects of nucleoside monophosphates on microalgae-bacteria symbiotic systems.

Microalgae are promising sources of clean energy. Bioflocculation by cocultured bacteria is an effective way to harvest microalgae. As a key foundation for microorganisms, phosphorus is theoretically effective in shaping microalgae production and flocculation. In this study, the impacts of 23 nucleoside monophosphates on Auxenochlorella pyrenoidosa growth, lipid synthesis, and self-settlement and on the symbiotic bacterial system were investigated. Adenosine monophosphate was the most effective in enhancing microalgae development (2.14-3.16 × 108 cells/mL) and lipid production (average 10.48%) and resulted in a low settling velocity. Samples were divided into two groups, purine and pyrimidine feeding, according to a random forest analysis (OOB = 0%, p < 0.001). Purine feeding resulted in the highest soluble extracellular protein and polysaccharide secretion (p < 0.01). KEGG ortholog count prediction of functional genes related to biofilm formation was conducted using PICRUSt2, and significant upregulation (FC ≥ 1.77, p < 0.05) of the extracellular polymeric substance formation functional group was observed in the adenosine and guanosine treatments. The symbiotic bacterial community structure differed substantially between purine- and pyrimidine-feeding systems. In summary, these results indicated that the effect of nucleoside monophosphates on the microalgae-bacteria system is determined by the base type (purine or pyrimidine) rather than the molecular structure (cyclic or noncyclic).

Nitrifiers Cooperate to Produce Nitrous Oxide in Plateau Wetland Sediments.

The thawing of dormant plateau permafrost emits nitrous oxide (N2O) through wetlands; however, the N2O production mechanism in plateau wetlands is still unclear. Here, we used the 15N-18O double tracer technique and metagenomic sequencing to analyze the N2O production mechanism in the Yunnan-Kweichow and Qinghai-Tibet plateau wetlands during the summer of 2020. N2O production activity was detected in all 16 sediment samples (elevation 1020-4601 m: 2.55 ± 0.42-26.38 ± 3.25 ng N g-1 d-1) and was promoted by nitrifier denitrification (ND). The key functional genes of ND (amoA, hao, and nirK) belonged to complete ammonia oxidizing (comammox) bacteria, and the key ND species was the comammox bacterium Nitrospira nitrificans. We found that the comammox bacterial species N. nitrificans and the ammonia oxidizing bacterial (AOB) species Nitrosomonas europaea cooperate to produce N2O in the plateau wetland sediments. Furthermore, we inferred that environmental factors (elevation and total organic matter (TOM)) influence the cooperation pattern via N. nitrificans, thus affecting the N2O production activity in the plateau wetland sediments. Our findings advance the mechanistic understanding of nitrifiers in biogeochemical cycles and global climate change.

Comparing with oxygen, nitrate simplifies microbial community assembly and improves function as an electron acceptor in wastewater treatment.

Biochemical oxidation and reduction are key processes in treating biological wastewater and they require the presence of electron acceptors. The functional impact of electron acceptors on microbiomes provides strategies for improving the treatment efficiency. This research focused on two of the most important electron acceptors, nitrate and oxygen. Molecule ecological network, null model, and functional prediction based on high-throughput sequencing were used to analyze the microbiomes features and assembly mechanism. The results revealed nitrate via the homogeneous selection (74.0%) decreased species diversity, while oxygen via the homogeneous selection (51.1%) and dispersal limitation (29.6%) increased the complexity of community structure. Microbes that were more strongly homogeneously selected for assembly included polyphosphate accumulating organisms (PAOs), such as Pseudomonas and variovorax in the nitrate impacted community; Pseudomonas, Candidatus_Accumulibacter, Thermomonas and Dechloromonas, in the oxygen impacted community. Nitrate simplified species interaction and increased the abundance of functional genes involving in tricarboxylic acid cycle (TCA cycle), electron transfer, nitrogen metabolism, and membrane transport. These findings contribute to our knowledge of assembly process and interactions among microorganisms and lay a theoretical basis for future microbial regulation strategies in wastewater treatment.