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The multi-soil-layering (MSL) systems is an emerging solution for environmentally-friendly and cost-effective treatment of decentralized rural domestic wastewater. However, the role of the seemingly simple permeable layer has been overlooked, potentially holding the breakthroughs or directions to addressing suboptimal nitrogen removal performance in MSL systems. In this paper, the mechanism among diverse substrates (zeolite, green zeolite and biological ceramsite) coupled microorganisms in different systems (activated bacterial powder and activated sludge) for rural domestic wastewater purification was investigated. The removal efficiencies performed by zeolite coupled with microorganisms within 3 days were 93.8% for COD, 97.1% for TP, and 98.8% for NH4+-N. Notably, activated sludge showed better nitrification and comprehensive performance than specialized nitrifying bacteria powder. Zeolite attained an impressive 89.4% NH4+-N desorption efficiency, with a substantive fraction of NH4+-N manifesting as exchanged ammonium. High-throughput 16S rRNA gene sequencing revealed that aerobic and parthenogenetic anaerobic bacteria dominated the reactor, with anaerobic bacteria conspicuously absent. And the heterotrophic nitrification-aerobic denitrification (HN-AD) process was significant, with the presence of denitrifying phosphorus-accumulating organisms (DPAOs) for simultaneous nitrogen and phosphorus removal. This study not only raises awareness about the importance of the permeable layer and enhances comprehension of the HN-AD mechanism in MSL systems, but also provides valuable insights for optimizing MSL system construction, operation, and rural domestic wastewater treatment.

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Integrated fixed-film activated sludge (IFAS) system, an improvement of the activated sludge process, combines the advantages of both attached sludge (AS) and suspended sludge (SS). This study aimed to fully decipher the roles of AS and SS in simultaneous N and P removal in an IFAS system through metagenomic analysis. It was found that AS contributed about 84.04%, 97%, and 95.12% to exogenous NO3--N reduction, endogenous NO3--N reduction, and endogenous NO2--N reduction, respectively. Compared with AS, SS exhibited a greater contribution to anaerobic P release (69.06%) and aerobic P uptake (73.48%). Nitrate and nitrite reductase enzymes showed higher activities in AS, while the activities of exopolyphosphatase and alkaline phosphatase D were more active in SS. P content further indicated that in AS, only a small amount of P was stored in EPS, with most presented intracellularly. In SS, the amount of P stored in EPS was found to be higher. Metagenomic analysis revealed genes related to the synthesis and degradation of endogenous carbon were higher in AS, whereas the TCA cycle exhibited higher activity in SS. P removal-related genes (such as ppk2, ppx, and adk) was significantly higher in SS than in AS. The alteration of genes associated with nitrogen metabolism suggested that the microbes in AS had a higher capacity for nitrification and denitrification. In summary, the discrepancy in the roles of AS and SS in N and P removal in IFAS can be attributed to variations in enzyme activity, P storage in EPS, microbial community composition, and functional gene abundance.

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Global climate change and rapid urbanization have resulted in more frequent and intense rainfall events in urban areas, raising concerns about the effectiveness of stormwater bioretention systems. In this study, we optimized the design by constructing a multi-layer filler structure, including plant layer, biochar layer, and pyrite layer, and evaluated its performance in nitrogen (N) and phosphorus (P) removal under different temperatures (5-18 °C and 24-43 °C), rainfall intensity (47.06 mm rainfall depth), and frequency (1-5 days rainfall intervals) conditions. The findings indicate that over 775 days, the plant system consistently removed 62.3% of total nitrogen (TN) and 97.0% of total phosphorus (TP) from 103 intense rainfall events. Temperature fluctuations had minimal impact on nitrate nitrogen (NO3--N) and TP removal, with differences in removal rates of only 1.0% and 0.6%, respectively, among plant groups. Across the multi-layer structure, plant roots mitigated the impact of temperature differences on NO3--N removal, while high-frequency rainfall fluctuated the stability of NO3--N removal. Dense plant roots reinforced N and P removal by facilitating denitrification in the vadose zone (biochar) and strengthening denitrification processes. Biochar and pyrite contributed to stable microenvironments and diverse ecological functions, enhancing NO3--N and PO43- removal. In summary, the synergistic effects of the multi-layer filler structure improved and stabilized N and P removal, providing valuable insights for addressing runoff pollution in bioretention systems amidst rapid urbanization and climate change challenges.

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Microbial consortia in riverbed substrates and their extracellular matrix (biofilms) play a key role in phosphorus (P) entrapment. When P entrapment saturates, the benthic compartment changes from a P sink to a P source thus increasing eutrophication risk. P entrapment saturation is expected to differ between intracellular and extracellular P entrapment and between different magnitudes and durations of P inputs. We studied biofilm P-entrapment following short (48 h) and long (14 days) P loading events in stream bypass flumes supplied with a gradient of dissolved P concentrations. This allowed us to link local biofilm processes in sediments to potential effects on river self-purification, via quantifying the P removal efficiency in the flumes. We found that in short-term events, biofilms develop intracellular mechanisms to cope with P inputs, while long-term events and high P inputs suppress the intracellular uptake mechanisms and increase the prevalence of extracellular entrapment. Specifically, long-term events lowered the threshold for intracellular P entrapment saturation, and decreased the ratio between intracellular and extracellular entrapment resulting in lower removal efficiency for dissolved phosphorus. Our results highlight the risk that aquatic ecosystems may face as the ratio of intracellular to extracellular P entrapment decreases, which may reduce their ability to deal with P inputs, thereby increasing risks of eutrophication.

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Phosphorus (P) pollution in aquatic environments poses significant environmental challenges, necessitating the development of effective remediation strategies, and biochar has emerged as a promising adsorbent for P removal at the cost of extensive research resources worldwide. In this study, a machine learning approach was proposed to simulate and predict the performance of biochar in removing P from water. A dataset consisting of 190 types of biochar was compiled from literature, encompassing various variables including biochar characteristics, water quality parameters, and operating conditions. Subsequently, the random forest and CatBoost algorithms were fine-tuned to establish a predictive model for P adsorption capacity. The results demonstrated that the optimized CatBoost model exhibited high prediction accuracy with an R2 value of 0.9573, and biochar dosage, initial P concentration in water, and C content in biochar were identified as the predominant factors. Furthermore, partial dependence analysis was employed to examine the impact of individual variables and interactions between two features, providing valuable insights for adsorbent design and operating condition optimization. This work presented a comprehensive framework for applying a machine learning approach to address environmental issues and provided a valuable tool for advancing the design and implementation of biochar-based water treatment systems.

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In recent years, with the global rise in awareness regarding carbon neutrality, the treatment of wastewater in rural areas is increasingly oriented towards energy conservation, emission reduction, low-carbon output, and resource utilization. This paper provides an analysis of the advantages and disadvantages of the current low-carbon treatment process of low-carbon treatment for rural wastewater. Constructed wetlands (CWs) are increasingly being considered as a viable option for treating wastewater in rural regions. In pursuit of carbon neutrality, advanced carbon-neutral bioprocesses are regarded as the prospective trajectory for achieving carbon-neutral treatment of rural wastewater. The incorporation of CWs with emerging biotechnologies such as sulfur-based autotrophic denitrification (SAD), pyrite-based autotrophic denitrification (PAD), and anaerobic ammonia oxidation (anammox) enables efficient removal of nitrogen and phosphorus from rural wastewater. The advancement of CWs towards improved removal of organic and inorganic pollutants, sustainability, minimal energy consumption, and low carbon emissions is widely recognized as a viable low-carbon approach for achieving carbon-neutral treatment of rural wastewater. This study offers novel perspectives on the sustainable development of wastewater treatment in rural areas within the framework of achieving carbon neutrality in the future.

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Membrane aerated biofilm reactor (MABR) is challenged by biofilm thickness control and phosphorus removal. Air scouring aided by computational fluid dynamics (CFD) was employed to detach outer biofilm in sequencing batch MABR treating low C/N wastewater. Biofilm with 177-285 µm thickness in cycle 5-15 achieved over 85 % chemical oxygen demand (COD) and total inorganic nitrogen (TIN) removals at loading rate of 13.2 gCOD/m2/d and 2.64 gNH4+-N/m2/d. Biofilm rheology measurements in cycle 10-25 showed yield stress against detachment of 2.8-7.4 Pa, which were equal to CFD calculated shear stresses under air scouring flowrate of 3-9 L/min. Air scouring reduced effluent NH4+-N by 10 % and biofilm thickness by 78 µm. Intermittent aeration (4h off, 19.5h on) and air scouring (3 L/min, 30 s before settling) in one cycle achieved COD removal over 90 %, TIN and PO43--P removals over 80 %, showing great potential for simultaneous carbon, nitrogen and phosphorus removals.

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The effectiveness of phosphorus (P) removal by sand filters is limited during septic tank effluent (STE) treatment. The elevated effluent P concentrations pose threats to drinking water quality and contribute to eutrophication. The concern of P leaching from sand filters is further exacerbated by the increased frequency of flooding and natural precipitation due to climate change. This study aimed to understand P attenuation and leaching dynamics, as well as the removal mechanisms in sand filters treating STE, offering insights into the design and implementation of P removal/recovery modules to onsite wastewater treatment systems. P attenuation and leaching during STE treatment and rainfall were studied in bench-scale columns (new vs. aged sand). At standard STE loading (1.2 gallon d-1 ft-2), 24-32% removal of total phosphorus (TP) was achieved, while increased P removal efficiency (35-53%) was observed at low loading (0.6 gallon d-1 ft-2) with influent containing 10.3-20.0 mg P L-1. Complete breakthroughs were observed in both aged (12-70 days) and new columns (27-73 days) at test hydraulic loadings. The maximum TP attenuation level was 20.6-45.3 mg P kg-1 and 25.3-33.0 mg P kg-1, in aged and new sand columns, respectively. When simulated rain was applied (15-60 mm h-1), 80-97% of the attenuated P leached out and the leaching dynamics were impacted by rainfall duration rather than the intensity. The highest concentrations of TP (15.6-15.9 mg L-1) were leached out from both columns within the first 2-6 h. Orthophosphate was the dominant P species in treated effluent (83-84%) and leachate (69-88%), demonstrating its significance as the major P form in the discharge. In addition, aged sand (>5 years) accumulated higher levels of Mg, Al, Ca, and Fe, thus enhancing the P attenuation level during STE treatment. Collectively, this study underscored the importance of frequent field monitoring for reliable long-term P removal estimates.

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A comparative study explored how photoaging, ozonation aging, and Fenton aging affect tire wear particles (TWPs) and their phosphorus (P) removal in activated sludge. Aging altered TWPs' properties, increasing surface roughness, porosity, and generating more small particles, especially environmental persistent free radicals (EPFRs) in ozonation and Fenton aging. Post-aging TWPs (50 mg/L) inhibited sludge P removal significantly (p < 0.05), with rates of 44.3% and 59.6% for ozonation and Fenton aging, respectively. In addition, the metabolites involved in P cycling (poly-ß-hydroxyalkanoates: PHA and glycogen) and essential enzymes (Exopolyphosphatase: PPX and Polyphosphate kinase: PPK) were significantly inhibited (p < 0.05). Moreover, TWPs led to a decrease in microbial cells within the sludge and altered the community structure, a situation exacerbated by the aging of TWPs. P-removing bacteria decreased (e.g., Burkholderia, Candidatus), while extracellular polymeric substance-secreting bacteria increased (e.g., Pseudomonas, Novosphingobium). Pearson correlation analysis highlighted EPFRs' role in TWPs' acute toxicity to microbial cells, yet, emphasizing particle size's impact on the sludge system's purification and community structure.

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Fungi capable of simultaneous nitrogen and phosphorus removal from wastewater is rarely found. Here, a novel fungal strain (SNDM1) performing heterotrophic nitrification, aerobic denitrification, and phosphate removal was isolated and identified as Mucor circinelloides. The favorable nutrient removal conditions by the strain using glucose were C/N ratios of 25-30, salinities of 0 %-3 %, and pH of 7.5. Strain SNDM1 achieved ammonium, nitrite, nitrate, and phosphate removal rates of 5.23, 10.08, 4.88, and 0.97 mg/L/h. Nitrogen balance indicated that gaseous (18.60 %-24.55 %) and intracellular nitrogen (43.76 %-70.63 %) were primary fate of initial nitrogen. Enzyme activity revealed that ammonium removal occurred through heterotrophic nitrification and aerobic denitrification. Removed phosphorus was mainly transformed into cell membranes (56 %-64 %) and extracellular polymeric substances (20 %-26 %). Orthophosphate was the major intracellular phosphorus species, while polyphosphate and pyrophosphate existed extracellularly. These findings highlight the potential of this fungal strain for bioremediating polluted wastewater.

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This study uncovered the response of novel micro-granule wastewater treatment technology to different powder carrier materials. Characteristics and distinctions among different systems were assessed based on process performance, sludge aggregation capacity, and microbial metabolism. Zeolite carrier system exhibited remarkable nitrogen removal efficiency of 89.6 ± 0.9 %, while diatomite carriers, in conjunction with intermittent aeration, enhanced simultaneous nitrification and denitrification from 2.6 % to 27.1 %. Iron-based carriers demonstrated efficient phosphorus removal (94.7 ± 1.2 %) through both chemical and microbial pathways. Specific surface area, pore structure and biocompatibility of powder carriers determined the formation and size of micro-granules. Tryptophan-like substances, C-(C/H), and Npr in extracellular polymeric substances strongly correlated with sludge hydrophobicity and granulation. Significant enrichment in norank_Comamonadaceae and Nitrosomonas in zeolite powder carrier system promoted partial nitrification and endogenous denitrification. Differences in metabolic pathways elucidated the up-regulation of amino acid synthesis, energy metabolism, and membrane transport as potential mechanisms driving micro-granule formation and efficient treatment performance.

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Denitrifying phosphorus removal (DPR), which is dominated by denitrifying polyphosphate-accumulating organisms (DPAOs), is a promising process for nitrogen and phosphorus removal. Denitrifying glycogen-accumulating organisms (DGAOs) and DPAOs typically coexist in the DPR sludge, complicating the study of DPAOs' denitrification capacity. In this study, two reactors were fed with nitrate and nitrite during the anoxic phase to cultivate nitrate-DPR and nitrite-DPR sludge. Both reactors yielded high and low DGAO abundance sludges, enabling the evaluation of the denitrification capacity of DPAOs. For the nitrate-DPR sludge, the nitrite reduction rate was 1.63 times higher than the nitrate reduction rate when DPAOs were the primary denitrifiers. For the nitrite-DPR sludge, the reduction rate of nitrite was more than three times that of nitrate, irrespective of DGAO abundance. These findings indicated that DPAOs preferred nitrite to nitrate and were well suited to reduce nitrite rather than reduce nitrate to supply nitrite.

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The denitrifying sulfur (S) conversion-associated enhanced biological phosphorus removal (DS-EBPR) process for treating saline wastewater is characterized by its unique microbial ecology that integrates carbon (C), nitrogen (N), phosphorus (P), and S biotransformation. However, operational instability arises due to the numerous parameters and intricates bacterial interactions. This study introduces a two-stage interpretable machine learning approach to predict S conversion-driven P removal efficiency and optimize DS-EBPR process. Stage one utilized the XGBoost regression model, achieving an R2 value of 0.948 for predicting sulfate reduction (SR) intensity from anaerobic parameters with feature engineering. Stage two involved the CatBoost classification and regression model integrating anoxic parameters with the predicted SR values for predicting P removal, reaching an accuracy of 94% and an R2 value of 0.93, respectively. This study identified key environmental factors, including SR intensity (20-45 mg S/L), influent P concentration (<9.0 mg P/L), mixed liquor volatile suspended solids (MLVSS)/mixed liquor suspended solids (MLSS) ratio (0.55-0.72), influent C/S ratio (0.5-1.0), anoxic reaction time (5-6 h), and MLSS concentration (>6.50 g/L). A user-friendly graphic interface was developed to facilitate easier optimization and control. This approach streamlines the determination of optimal conditions for enhancing P removal in the DS-EBPR process.

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Tetracycline and copper ion are common pollutants in wastewater, and the effects of mixed pollutants on microorganisms in wastewater biological treatment have been less studied. In order to reveal the effects of mixed pollutants of tetracycline and copper ion on the microorganisms during the biological phosphorus removal, three ratios of tetracycline and copper ions were designed by the direct equipartition ray method. The relative abundance and diversity of microbial community were investigated, and the microbial interactions were revealed through microbiological methods. The results demonstrated that, for three different ratios, the inhibitory effect of specific phosphorus uptake rate became more significant with the increase of the tetracycline-copper ions concentration and the reaction time. The microbial community decreased with the increase of the proportion of tetracycline in different ratios. The relative abundance of Acinetobacter decreased with the increase of the proportion of tetracycline, while the relative abundance of Ca.Competibacter was higher under the conditions of low mixtures concentrations. Positive interactions and symbiotic relationships among microorganisms were predominant for three different ratios. However, as the proportion of tetracycline increased, the community structure of microorganisms shifted from phosphate-accumulating organisms to glycogen accumulating organisms and denitrifying bacteria. This study can provide a reference for the effect of mixed pollutants on microorganisms and the mechanism of wastewater treatment.

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Glycerol, an abundant by-product of biodiesel production, represented a promising carbon source for enhancing nutrient removal from low C/N ratio wastewater. This study discovered a novel approach to initiate glycerol-driven denitrifying phosphorus removal (DPR) in situ by creating a short-term microaerobic environment within the aerobic zone. This approach facilitated the in-situ conversion of glycerol, which was subsequently utilized by denitrifying phosphate accumulating organisms (DPAOs) for DPR. The feasibility and stability of glycerol-driven DPR were validated in a continuous-flow pilot-scale reactor. Anaerobic phosphorus release increased from 1.0 mg/L/h to 2.5 mg/L/h, with fermentation bacteria and related functional genes showing significant increases. The stable stage exhibited 92.8% phosphorus removal efficiency and 55.5% DPR percentage. The microaerobic environment enhanced fermentation bacteria enrichment, crucial for glycerol-driven DPR stability. The collaborative interaction between fermentation bacteria and phosphate accumulating organisms (PAOs) played a key role in sustaining glycerol-driven DPR stability. These findings provide a robust theoretical foundation for applying glycerol-driven DPR in established wastewater treatment plants.

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Despite the potential of sodium sulfide (Na2S) for phosphorus (P) recovery from iron-phosphate waste, the underlying mechanism regarding its impact on P conversion and product quality has not been well addressed. In this study, the effects of Na2S addition on P release and recovery from a chemical-enhanced phosphorus removal (CEPR) sludge during anaerobic fermentation were systematically investigated. The results revealed that the effective mobilization of P bound to Fe (Fe-P) by Na2S dominated the massive P release from the CEPR sludge, while the organic P (OP) release was not significantly enhanced during anaerobic fermentation. Due to the rapid reaction of Na2S with Fe-P and the prevention of Fe(II)-P precipitation by excess S2-, the Fe-P was decreased by 9.7%, 15.2% and 24.9% at S:Fe molar ratios of 0.3, 0.5 and 1, respectively. After anaerobic fermentation, the released P mainly existed as soluble phosphate (SP), P bound to Ca (Ca-P) and P bound to Al (Al-P). The nitrogen and P contents in the fermentation supernatant significantly increased with higher S:Fe ratios, facilitating the efficient recovery of P as high-purity struvite. However, the increased Na2S dosage deteriorated the sludge dewaterability because of the dissolution of hydrophilic extracellular polymeric substances and the looser secondary structure of proteins. Comprehensively considering the P recovery, sludge dewaterability and economic cost, the optimal Na2S dosage was determined at the S:Fe ratio of 0.3. These findings provide novel insights into the role of Na2S in P recovery as struvite from CEPR sludge.

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This study developed a microelectrolysis-integrated constructed wetland with pyrite filler around the cathode (e-PCW) to treat eutrophic water. Results indicated that e-PCW effectively enhanced pyrite dissolution, converting solid-phase electron donors into bioavailable forms, thereby facilitating the enrichment of various denitrifying bacteria on pyrite surfaces. Importantly, iron-reducing and sulfur-reducing bacteria attached to the pyrite surfaces enhanced the conversion of ferric iron and sulfate, thereby driving iron and sulfur cycles and promoting electron transfer. Therefore, synergistic effects of pyrite and microelectrolysis made e-PCW achieve higher total nitrogen (TN) and total phosphorus (TP) removal efficiencies. With a hydraulic retention time of 24 h, the highest removal efficiencies of TN and TP achieved 78% and 75%, respectively. Furthermore, when eutrophic water containing high concentration of algae was fed into e-PCW, it consistently demonstrated superior TN and TP removal capabilities. This work provides a valuable approach to optimizing constructed wetland technology for treating eutrophic water.

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Introduction: Simultaneous chemical phosphorus removal process using iron salts (Fe(III)) has been widely utilized in wastewater treatment to meet increasingly stringent discharge standards. However, the inhibitory effect of Fe(III) on the biological phosphorus removal system remains a topic of debate, with its precise mechanism yet to be fully understood. Methods: Batch and long-term exposure experiments were conducted in six sequencing batch reactors (SBRs) operating for 155 days. Synthetic wastewater containing various Fe/P ratios (i.e., Fe/P = 1, 1.2, 1.5, 1.8, and 2) was slowly poured into the SBRs during the experimental period to assess the effects of acute and chronic Fe(III) exposure on polyphosphate-accumulating organism (PAO) growth and phosphorus metabolism. Results: Experimental results revealed that prolonged Fe(III) exposure induced a transition in the dominant phosphorus removal mechanism within activated sludge, resulting in a diminished availability of phosphorus for bio-metabolism. In Fe(III)-treated groups, intracellular phosphorus storage ranged from 3.11 to 7.67 mg/g VSS, representing only 26.01 to 64.13% of the control. Although the abundance of widely reported PAOs (Candidatus Accumulibacter) was 30.15% in the experimental group, phosphorus release and uptake were strongly inhibited by high dosage of Fe(III). Furthermore, the abundance of functional genes associated with key enzymes in the glycogen metabolism pathway increased while those related to the polyphosphate metabolism pathway decreased under chronic Fe(III) stress. Discussion: These findings collectively suggest that the energy generated from polyhydroxyalkanoates oxidation in PAOs primarily facilitated glycogen metabolism rather than promoting phosphorus uptake. Consequently, the dominant metabolic pathway of communities shifted from polyphosphate-accumulating metabolism to glycogen-accumulating metabolism as the major contributor to the decreased biological phosphorus removal performance.

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This study investigated the nutrient removal and microbial community succession in moving bed biofilm reactor under stable and three levels of influent carbon/nitrogen (C/N) ratio fluctuation (± 10%, ± 20%, and ± 30%). Under the conditions of influent C/N ratio fluctuation, the removal efficiency of COD and PO43--P decreased 4.7-6.4% and 3.7-12.9%, respectively, while the nitrogen removal was almost unaffected. A sharp decrease in the content of culturable functional bacteria related to nitrogen and phosphorus removal including nitrite-oxidizing bacteria (NOB), aerobic denitrifying bacteria (DNB), and polyphosphate-accumulating organisms (PAOs) from the carrier biofilm was observed. Sequencing analysis revealed that the abundance of Candidatus Competibacter increased 10.3-25.9% and became the dominant genus responsible for denitrification, potentially indicating that nitrate was removed via endogenous denitrification under the influent C/N ratio fluctuation. The above results will provide basic data for the nutrient removal in decentralized wastewater treatment under highly variable influent conditions.

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The effects of antibiotics, such as tetracycline, sulfamethoxazole, and ciprofloxacin, on functional microorganisms are of significant concern in wastewater treatment. This study observed that Acinetobacter indicus CZH-5 has a limited capacity to remove nitrogen and phosphorus using antibiotics (5 mg/L) as the sole carbon source. When sodium acetate was supplied (carbon/nitrogen ratio = 7), the average removal efficiencies of ammonia-N, total nitrogen, and orthophosphate-P increased to 52.46 %, 51.95 %, and 92.43 %, respectively. The average removal efficiencies of antibiotics were 84.85 % for tetracycline, 39.32 % for sulfamethoxazole, 18.85 % for ciprofloxacin, and 23.24 % for their mixtures. Increasing the carbon/nitrogen ratio to 20 further improved the average removal efficiencies to 72.61 % for total nitrogen and 97.62 % for orthophosphate-P (5 mg/L antibiotics). Additionally, the growth rate and pollutant removal by CZH-5 were unaffected by the presence of 0.1-1 mg/L antibiotics. Transcriptomic analysis revealed that the promoted translation of aceE, aarA, and gltA genes provided ATP and proton -motive forces. The nitrogen metabolism and polyphosphate genes were also affected. The expression of acetate kinase, dehydrogenase, flavin mononucleotide enzymes, and cytochrome P450 contributed to antibiotic degradation. Intermediate metabolites were investigated to determine the reaction pathways.

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