Search Results (96)

Partial nitrification (PN) is a promising strategy for reducing aeration demand and improving the energy efficiency of biological nitrogen removal in wastewater treatment. However, maintaining stable PN in continuous-flow activated sludge reactors remains challenging due to the recovery of nitrite-oxidizing bacteria (NOB) and the absence of cyclic operational phases that naturally promote microbial selectivity in sequencing batch reactors. This study proposes a model-based multi-criteria optimization framework to identify and classify feasible operating conditions for stable PN in continuous-flow activated sludge reactors. A modified Activated Sludge Model No. 1 (ASM1) was used to describe the dynamics of ammonia-oxidizing bacteria, nitrite-oxidizing bacteria, and heterotrophic biomass, while equilibrium points were determined through steady-state optimization and evaluated using a multi-criteria feasibility analysis based on nitrite accumulation ($\beta$), ammonium oxidation efficiency ($\alpha$), oxygen uptake rate (OUR), hydraulic retention time (HRT), and sludge retention time (SRT). Seasonal variability was incorporated through summer and winter operating scenarios. Results indicate that stable PN can be achieved under operating conditions of pH 7.5–8.5, dissolved oxygen concentrations between 0.3 and 2.5 mg/L, HRT values of approximately 2–3 h, and SRT values between 10 and 20 d. Under these conditions, high nitrite accumulation ($\beta >0.8$) and ammonium oxidation efficiency ($\alpha >0.8$) were maintained with moderate oxygen demand, although seasonal differences revealed greater operational flexibility in summer and tighter constraints in winter. The proposed framework provides a systematic approach for identifying robust and energy-efficient operating regions in continuous-flow PN systems and establishes a foundation for future supervisory control implementation in full-scale wastewater treatment applications. The study also shows that over 40% energy savings could be achieved at optimal equilibrium points for partial nitrification compared to full nitrification. Full article

Ammonium nitrogen pollution presents a significant challenge in wastewater treatment. Traditional activated sludge processes often suffer from limitations such as low efficiency and high energy consumption when treating high-ammonium nitrogen wastewater. This study utilized previously screened high-efficiency heterotrophic nitrification aerobic denitrification (HN-AD) bacterial strains (Pseudomonas alcaliphila and Paracoccus versutus) synergistically with microalgae to construct an immobilized bacteria algae symbiotic system (IBAS). The nitrogen removal performance and microbial community response of the system were investigated under different nitrogen sources, carbon to nitrogen (C/N) ratios, and light intensities. Results demonstrated that the system achieved a removal rate of over 95% for nitrite and nitrate. Under conditions of C/N = 15 and high light intensity (335.36 μmol/(m² · s)), the removal rates of NH₄⁺-N, TN, and COD exceeded 90% without nitrite accumulation. Microbial community analysis revealed that high C/N conditions significantly enriched HN-AD functional bacteria (such as Acinetobacter) in the Pseudomonadota phylum and Gammaproteobacteria class. High light intensity promoted the proliferation of microalgae (Chlorella and Halochlorella), enhanced algal bacterial interaction, and improved system stability. This study elucidated the nitrogen removal mechanism of the IBAS under multi-factor regulation, providing a theoretical foundation and demonstrating application potential for low-carbon and high-efficiency wastewater treatment technologies. Full article

To address the bottleneck of poor biological nitrogen removal efficiency caused by the extremely low carbon-to-nitrogen (C/N) ratio of domestic sewage in alpine plateau regions, this study used Hordeum vulgare var. coeleste L., a characteristic crop endemic to the Qinghai–Tibet Plateau, as raw material and adopted pretreated highland barley straw as an external carbon source. Three parallel experiments were carried out using the anaerobic–aerobic–anoxic sequencing batch reactor (AOA-SBR) process to investigate the nitrogen removal performance and functional succession of the microbial community in the AOA-SBR system under three C/N ratio ranges: 5~7, 7~9, and 9~11. The results showed that the addition of an external carbon source significantly improved nitrogen removal efficiency. The optimal C/N ratio range for nitrogen removal in this study was determined to be 7~9. A weakly alkaline environment was conducive to denitrification. The fermentation broth prepared by alkali pretreatment contained a large amount of readily biodegradable organic matter with low toxicity, and achieved excellent nitrogen removal performance, helping to realize cost reduction and efficiency improvement in wastewater treatment. At the optimal C/N ratio of 7~9, the average removal efficiencies of ammonia nitrogen (NH₄⁺-N) and total nitrogen (TN) reached 94.46% and 61.32%, respectively, which were significantly improved compared with the blank control group without external carbon addition. During the experimental period, no obvious changes were observed in microbial abundance at the phylum level, whereas the community structure at the genus level responded significantly to the addition of a straw carbon source. Among them, genera with specific degradation capabilities for straw hydrolysates, such as norank_f__Chitinophagaceae and unclassified_f__Comamonadaceae, were highly sensitive to variations in the C/N ratio. These genera could partially replace the nitrification and denitrification functions of other microorganisms and played a key role in the nitrogen removal process. In contrast, Thauera, a typical conventional heterotrophic denitrifier, showed no significant response to changes in the C/N ratio, indicating that the straw-based external carbon source mainly affected microbial genera with specific hydrolysate-degrading functions. Full article

Although numerous studies have focused on the potential application of heterotrophic nitrification–aerobic denitrification (HNAD) bacteria in wastewater treatment, research exploring their potential in aquaculture biofloc systems remains limited. In this study, a promising HNAD strain, identified as Stutzerimonas stutzeri MJ20, was isolated from mature biofloc. This strain efficiently utilized low-cost carbon sources (e.g., glucose) and small-molecule carbon sources (e.g., sodium acetate and sodium succinate). Under conditions with glucose as the carbon source, a carbon-to-nitrogen (C/N) ratio of 15, pH 6–9, temperature 25–35 °C, salinity 0–35‰, and shaker speed of 0–150 rpm, it achieved removal rates of 95–100% for NH₄⁺-N, NO₂⁻-N, and NO₃⁻-N at initial concentrations of 100 mg/L each. Even at higher concentrations (up to 200 mg/L NH₄⁺-N and 500 mg/L for both NO₂⁻-N and NO₃⁻-N), removal rates exceeded 99%. Under mixed nitrogen sources, strain MJ20 demonstrated efficient nitrogen removal, preferentially utilizing NH₄⁺-N, with only minimal and transient accumulation of nitrite and nitrate. Genomic analysis revealed that MJ20 carries key denitrification genes, including napA, nirS, norB and nosZ, and possesses complete pathways for nitrate reduction to nitrogen gas and ammonia assimilation, although typical autotrophic nitrification genes were not detected. Combined genomic data and autotrophic culture experiments indicated that, in addition to utilizing various organic carbon sources, the strain also exhibited certain autotrophic growth capabilities. Furthermore, MJ20 showed strong flocculation ability (flocculation rate > 96% within 16 h), sensitivity to multiple common antibiotics, and no toxicity to zebrafish, demonstrating favorable biosafety. In simulated seawater aquaculture wastewater with a C/N ratio of 5, it achieved a total nitrogen removal rate exceeding 94% within 72 h. These results indicate that strain MJ20 possesses comprehensive advantages, including efficient nitrogen removal, broad carbon source adaptability, strong environmental resilience, minimal accumulation of intermediate nitrogen products, excellent flocculation ability, and high biosafety. These traits highlight its potential for application in biofloc systems and in treating aquaculture tail water with a low C/N ratio. This study provides theoretical insights and practical guidance for screening HNAD bacteria suitable for biofloc systems. Full article

Deammonification, which is based on partial nitritation and anammox (PN/A), is a well-established sidestream treatment for nitrogen removal. However, transferring deammonification to mainstream wastewater treatment remains challenging due to low temperatures, the need to retain slow-growing anammox bacteria (AnAOB), and their competition for nitrite with nitrite-oxidizing bacteria (NOB) and heterotrophic denitrifiers. This work investigates cubic polyurethane foam carriers to promote growth and retention of AnAOB. A moving bed biofilm reactor (MBBR) and an integrated fixed-film activated sludge (IFAS) reactor were compared over a three-year experimental period at lab-scale. The feasibility of the biofilm carriers for deammonification was first evaluated under sidestream conditions, followed by a stepwise transition to mainstream operational conditions. The impact of operational parameters, including dissolved oxygen concentration, pH value, and aeration strategy, was evaluated with respect to the activity of aerobic ammonium-oxidizing bacteria (AOB), NOB, and AnAOB, as well as nitrogen removal rates. Deammonification reached nitrogen removal rates of 0.04–0.12 kg N m⁻³ d⁻¹ (IFAS reactor) and 0.02–0.28 kg N m⁻³ d⁻¹ (MBBR) at subphases with reactor bulk concentrations above 60 mg NH₄-N L⁻¹. Highest nitrogen removal degrees of 77 ± 6% (IFAS) and 76 ± 5% (MBBR) were achieved at reactor bulk concentrations of 96 mg NH₄ L⁻¹ and 97 mg NH₄ L⁻¹, respectively. Lower concentrations triggered NOB activity in both reactors, leading to an increase in nitrate concentration up to 22 mg NO₃-N L⁻¹. AOB and AnAOB activities were on average 6-fold higher on the carriers compared to suspended biomass throughout all experimental phases, demonstrating the feasibility of using cubic polyurethane foam carriers for deammonification. This was also confirmed by fluorescence in-situ hybridization (FISH) measurements. Median nitrogen removal rates over all experimental phases of 0.07 kg N m⁻³ d⁻¹ for the IFAS reactor and 0.05 kg N m⁻³ d⁻¹ for the MBBR were achieved, which are comparable to conventional activated sludge systems performing nitrogen removal via nitrification–denitrification. While at lower nitrogen concentrations, the IFAS reactor yielded superior nitrogen removal rates, peak nitrogen removal rates of 0.28 kg N m⁻³ d⁻¹ were measured in the MBBR configuration. However, controlling NOB activity at lower temperatures and concentrations remains a challenge in MBBR and IFAS configurations. In our study, in the IFAS reactor NOB activities were visible on fewer days than in MBBR. At mainstream-like conditions, higher nitrogen removal rates of IFAS (0.09–0.12 kg N m⁻³ d⁻¹) were achieved compared to the MBBR (0.06–0.09 kg N m⁻³ d⁻¹). This demonstrates the advantage of the IFAS reactor in treating mainstream wastewater via deammonification. As an autotrophic nitrogen removal process, the implementation of deammonification in the mainstream of municipal wastewater treatment plants enables enhanced recovery of biogas from sewage organic matter. The latter would otherwise be consumed during the conventional nitrification-denitrification pathway. Consequently, the overall energy balance for wastewater treatment can be improved, contributing to a more environmentally sustainable process. Full article

In Northeast China’s degraded croplands, nitrate (NO₃⁻-N) leaching is the dominant pathway for fertilizer-nitrogen (N) loss, which presents an increasing threat to the quality of groundwater. Conservation tillage, defined as no-tillage (NT) and straw retention, is a widely adopted management strategy to maintain cropland fertility in the black soil (BS) regions. At present, however, the impact of shifting from conventional to conservation tillage on the vertical distribution and regulatory mechanisms of NO₃⁻-N derived from applied fertilizer-N (F_NO3) remains poorly understood. Based on a 12-year field experiment, we integrated ¹⁵N-tracing field monitoring with ¹⁵N-paired-labeling incubation to quantify the vertical migration of F_NO3 into deep soil profiles, and specify the dominant processes regulating N retention and supply. Across the tested BS croplands, total NO₃⁻-N production rates (4.06–6.58 mg N kg⁻¹ soil day⁻¹) were faster than their consumption rates (0.36–0.92 mg N kg⁻¹ soil day⁻¹), leading to a net accumulation of NO₃⁻-N, and implying a potential for leaching of NO₃⁻-N, from the perspective of substrate availability. The results of the field ¹⁵N micro-plot experiment also indicated that, by maize maturity in the first growing season, an average of 7.5% of F_NO3 had migrated to the 80–100 cm soil layer. During the following two growing seasons, the maximum accumulation of F_NO3 had shifted downward to 140–160 cm and 180–220 cm, respectively. Such a pattern, particularly in light of the increased extreme precipitation in the studied regions, raises clear concerns about NO₃⁻-N leaching losses. Compared with conventional management, no-tillage with full-rate straw mulching decreased net rates of NO₃⁻-N production from 6.22 to 3.14 mg N kg⁻¹ soil day⁻¹. This reduction resulted from a decline in the gross oxidation of NH₄⁺-N to NO₃⁻-N (from 6.39 to 3.70 mg N kg⁻¹ soil day⁻¹) and an increase in DNRA (from 0.35 to 0.85 mg N kg⁻¹ soil day⁻¹), which collectively delayed the downward transport of F_NO3. Conservation tillage also increased the gross rate of heterotrophic nitrification (from 0.19 to 0.36 mg N kg⁻¹ soil day⁻¹) and its proportion relative to total nitrification (from 2.8% to 8.9%). Despite this shift, autotrophic nitrification remained the dominant process for NO₃⁻-N production in the tested BS croplands, likely due to a pH constraint on heterotrophic nitrification. With the increasingly widespread promotion of conservation tillage for soil fertility improvement, heterotrophic nitrification warrants greater attention, particularly in BS regions where pH < 6.5 and C/N contents are relatively high. Collectively, our findings provide a scientific basis for tailoring tillage practices to maintain sustainable agriculture in Northeast China. Full article

Research on heterotrophic bioflocs is extensive, whereas investigations into autotrophic bioflocs remain limited. This study established an in situ autotrophic biofloc (ABF) system for intensive Pacific white shrimp (Penaeus vannamei) farming, aiming for zero water exchange and optimized water quality. A 120-day indoor experiment tested three stocking densities (300 (T1), 250 (T2), and 200 shrimp per m³ (T3)) with no water exchange. Water quality was monitored every two days, and bacterial communities were analyzed on days 10 and 70. The results indicated that ABF maturation was achieved by day 70 across all treatments, marked by three key indicators: (1) synchronous declines in nitrite and nitrate concentrations; (2) concurrent decreases in pH and total alkalinity approaching maturation; and (3) sustained high nitrogen removal efficiency (nitrite < 0.7 mg/L, ammonia < 0.6 mg/L). All density groups displayed similar patterns in both water quality dynamics and microbial community evolution. Bacterial analysis revealed that dominant genera such as Ruegeria, Bacillus, Muricauda, SM1A02, and Nitrospira played critical roles in toxic nitrogen removal, while pathogenic Klebsiella and Vibrio significantly decreased post-maturation. Heterotrophic nitrification and aerobic denitrification microorganisms (HNADMs) were identified as potentially responsible for nitrite accumulation. Nitrite accumulation was found in all groups. T2 and T3 achieved satisfactory breeding performance despite pre-maturation nitrate peaks exceeding 40 mg/L, whereas T1 suffered a low survival rate (27.47%) due to severe nitrite accumulation (>50 mg/L). A biofloc volume (BFV) of 4–8 mL/L effectively managed daily feed inputs of 75–110 g/m³. These findings lay a theoretical and technical foundation for the application of in situ ABF cultivation in intensive farming and enhance the sustainability of aquaculture. Full article

Nitrogen pollution in wastewater remains a pressing environmental concern, prompting the need for efficient and sustainable treatment technologies. This study constructs an HN-AD microbial consortium using three pre-screened strains (Delftia tsuruhatensis SDU2, Pseudomonas stutzeri SDU10, Alcaligenes faecalis SDU20) and evaluates its bioaugmentation effects in sequencing batch reactors (SBRs). An orthogonal test optimized the inoculation ratio as 2:3:3 (SDU2:SDU10:SDU20), achieving the highest ammonium removal efficiency of 96.02% in vitro. In SBR experiments, the bioaugmented reactor (SBR1) demonstrated superior nitrogen and organic matter removal compared to the control (SBR2). By day 40 of the enhancement phase, SBR1 achieved 88.9% ammonium removal and 93.7% COD removal, representing improvements of 20.5% and 17.9% over SBR2, respectively. Microbial community analysis revealed that bioaugmentation enriched Proteobacteria and Bacteroidota phyla, promoting functional guilds like Alcaligenes, Pseudomonas, and Paracoccus, which synergized with indigenous microbiota to enhance metabolic efficiency. This study highlights the potential of HN-AD consortia to overcome limitations of conventional nitrogen removal systems, offering a promising strategy for optimizing wastewater treatment processes. Full article

Heterotrophic nitrification and aerobic denitrification (HN–AD) is an emerging biological process capable of achieving efficient nitrogen removal in a single reactor. This study investigates the HN–AD performance of a sequencing batch reactor (SBR) operated with a simple anaerobic–aerobic cycle for treating high C/N wastewater. Over a 220-day operation, the system achieved average removal efficiencies of 98.6% for COD, 93.3% for NH₄⁺-N, and 87.1% for total nitrogen. Effluent concentrations of NO₂⁻-N and NO₃⁻-N remained negligible at the end of each aerobic phase. Concentration profiles of NH₄⁺-N, NO₂⁻-N, and NO₃⁻-N throughout the operation cycles confirmed the occurrence of simultaneous nitrification and aerobic denitrification. The consistently high COD removal and robust nitrogen reduction highlight the stability of the HN–AD microbial consortia enriched from activated sludge. Phosphorus removal (average removal efficiency 66.3%) may be enhanced by increasing the activity of phosphate-accumulating organisms (PAOs) through process optimization. This study demonstrated effective HN–AD using activated sludge in SBRs. Future work will focus on evaluating the system with real wastewater and continuous-flow setups to further refine operational parameters for sustained HN–AD performance. Full article

Municipal wastewater with a low carbon-to-nitrogen (C/N) ratio presents challenges for conventional nitrogen removal processes, often requiring costly external carbon sources. This study investigated the enhancement of nitrogen removal in a simultaneous nitrification and denitrification (SND) system by incorporating heterotrophic nitrification and aerobic denitrification (HN-AD) bacterial agents (Klebsiella variicola L3, Acinetobacter beijerinckii W4, and Acinetobacter sp. Z1) with modified basalt fiber carriers. Three reactors were compared: mixed HN-AD strains (M), mixed strains with activated sludge (A+M), and activated sludge alone (A). Results demonstrated that the A+M reactor achieved superior performance, with median removal efficiencies of 82.2% for NH₄⁺-N, 52.9% for total nitrogen (TN), and 51.6% for COD, outperforming the M reactor (75.2%, 43.6%, and 51.6%) and the A reactor (63.2%, 29.3%, and 44.8%). The A+M reactor also exhibited a 40% reduction in COD consumption per unit TN removed (2.55 ± 1.75) compared to the control reactor A (4.25 ± 3.99). Microbial analysis revealed Acinetobacter sp. Z1 (6.1%) and K. variicola L3 (1.1%) as dominant species, with the A+M reactor showing higher microbial diversity (56.4% Proteobacteria, 10.2% Bacteroidota) and biological viability (VSS/SS ratio of 0.70 ± 0.01). Extracellular polymeric substance (EPS) content in A+M reached 242.26 ± 15.52 mg/g-VSS, with a protein-to-polysaccharide ratio of 2.77 ± 0.00, indicating robust biofilm activity. These findings highlight the potential of HN-AD bacterial agents to enhance nitrogen removal in low C/N wastewater treatment, offering a cost-effective and sustainable alternative to traditional methods by reducing reliance on external carbon sources and improving system efficiency. Full article

Aerobic denitrifying microorganisms, with their strong environmental adaptability, low dissolved oxygen concentration requirements, rapid growth rate, and high nitrogen removal efficiency, significantly compensate for the shortcomings of traditional aerobic chemolithoautotrophic nitrification and anaerobic heterotrophic denitrification models. The introduction of aerobic denitrifiers can effectively enhance the removal of nitrate nitrogen. However, directly inoculating aerobic denitrifiers into wastewater leads to issues such as easy loss of bacterial cells and difficulty in forming a dominant flora, thus preventing the long-term maintenance of their enhancing effect on denitrification performance. To address this problem, microbial immobilization technology has been introduced into the remediation process of nitrogen-polluted water bodies. This technology can maintain a high biomass concentration, provide a stable breeding ground for microorganisms, and effectively prevent the rapid loss of microorganisms. This article systematically reviews the current status of the isolation of aerobic denitrifying bacteria, key enzymes, and genes, as well as the application progress of aerobic denitrifying bacteria and their immobilization technology, aiming to provide solid theoretical support for the practical application of aerobic denitrification technology and promote its further development in the field of nitrogen pollution control. Full article

Linear anionic surfactants (LAS) pose significant stress to microbial denitrification in wastewater treatment. This study investigated the performance and adaptation mechanisms of heterotrophic nitrification-aerobic denitrification (HN-AD) microbial consortia under LAS exposure after short-term (SCM, 2 months) and long-term (LCM, 6 months) acclimation. Results showed a dose-dependent inhibition of total nitrogen (TN) removal, with LCM achieving 97.40% TN removal under 300 mg/L LAS, which was 16.89% higher than SCM. Biochemical assays indicated that LCM exhibited lower reactive oxygen species (ROS) levels, a higher ATP content, and reduced LDH release, suggesting enhanced oxidative stress resistance and membrane stability. EPS secretion also increased in LCM, contributing to environmental tolerance. Metagenomic analysis revealed that long-term acclimation enriched key genera including Pseudomonas, Aeromonas, and Stutzerimonas, which maintained higher expression of denitrification (e.g., nosZ, nirS) and ammonium assimilation genes (glnA, gltB). Although high LAS concentrations reduced overall community diversity and led to convergence between SCM and LCM structures, LCM retained greater functional capacity and stress resistance. These findings underscore the importance of acclimation in sustaining denitrification performance under surfactant pressure and offer valuable insights for engineering robust microbial consortia in complex wastewater environments. Full article

This study investigates the enhancement of nitrogen removal performance in an intermittent biological sponge iron system (BSIS) through the immobilization of aerobic denitrifying bacteria. The aim is to improve the efficiency of simultaneous nitrification and denitrification (SND) in the BSIS by optimizing the microbial community involved in nitrogen conversion. The immobilization technique not only stabilizes the microbial activity and abundance of aerobic denitrifying bacteria, but also promotes a more efficient denitrification process. The optimal material ratio of polyvinyl alcohol–sodium alginate gel beads was determined as 10 g/100 mL PVA, 4 g/100 mL SA, 2 g/100 mL CaCl₂, and 2 g/100 mL of bacterial suspension, achieving a maximum NO₃⁻-N removal rate of 91.73%. A response surface model (RSM), established for the operational conditions, (shaker speed, temperature, and pH) showed a high fitting degree (R2 = 0.9960) and predicted the optimal conditions for maximum NO₃⁻-N removal as 109.24 rpm, 23.6 °C, and pH 7.9. Compared to R1 (47.82%), R3 achieved a higher average total nitrogen (TN) removal rate of 95.49%, following the addition of immobilized aerobic denitrifying bacteria to the BSIS. Full article

Nitrogen removal in the sewage treatment process is a significant challenge. The increase in nitrogen content in sewage leads to the eutrophication of water bodies and the deterioration of water quality in polluted environments. Therefore, converting nitrogen into non-polluting gases is a crucial and essential part of the sewage treatment process. Compared to physical, chemical, and physicochemical methods, biological denitrification is not only simple to operate and economically effective but also has less secondary pollution and saves energy. This paper summarizes the latest research progress on mainstream biological denitrification technology in WWTPS (wastewater treatment plants) and discusses its research background, methodology, and challenges. It is noted that the traditional biological nitrogen removal method is stable and widely used, but it has drawbacks such as high costs and long reaction times, especially in high-nitrogen-load wastewater treatment where its effectiveness is limited. The short-cut nitrification–denitrification process suits high-nitrogen-loading and a low C/N ratio wastewater as it reduces carbon source consumption. However, the problems of water quality fluctuation and unstable dissolved oxygen still need to be solved. The anaerobic ammonia oxidation process efficiently converts ammonia and nitrite to nitrogen using anaerobic ammonia-oxidizing bacteria, consuming less energy but facing limitations due to slow bacterial growth rates and stringent environmental conditions. The heterotrophic nitrification–aerobic denitrification process merges the traits of heterotrophic nitrifying bacteria and aerobic denitrifying bacteria, effectively reducing the ecological footprint and enhancing treatment efficiency. This approach is a pivotal focus for future research endeavors. Full article

Managing nitrogen and phosphorus pollution in high-salinity wastewater is a critical challenge for sustainable aquaculture and environmental protection. In this study, a novel salt-tolerant strain, Pseudomonas sediminis D4, was isolated from a brackish water environment. This strain exhibited simultaneous heterotrophic nitrification–aerobic denitrification and phosphorus removal capabilities. Biosafety assays demonstrated that the strain was antibiotic-sensitive and safe for aquatic environments. The optimal conditions for nitrogen and phosphate removal of strain D4 were carbon/nitrogen (C/N) ratio 10, phosphorus/nitrogen (P/N) ratio 0.2, pH 7, and temperature 30 °C while using sodium succinate as the carbon source. Under these conditions, strain D4 achieved removal efficiencies of 97.36% for ammonia (NH₄⁺-N), 100.00% for nitrate (NO₃⁻-N), and 98.02% for nitrite (NO₂⁻-N), along with 94.69%, 89.56%, and 97.40% removal of PO₄³⁻P, respectively. The strain exhibited strong salinity tolerance, functioning effectively within a range of 0% to 5% (w/v), and maintaining high nitrogen and phosphorus removal efficiency at a salinity of 3%. Enzyme activity assays verified the existence of key enzymes, such as ammonia nitrogen oxidase, nitrate oxidoreductase, nitrate reductase, nitrite reductase, polyphosphate kinase, and exopolyphosphatase, which are essential for the heterotrophic nitrification-aerobic denitrification and phosphorus removal capabilities of D4. These findings highlight the potential of Pseudomonas sediminis D4 for the biological treatment of high-salinity wastewater. Full article