Search

Simultaneous nitrification-denitrification (SND) is a promising nitrogen removal process. However, total nitrogen (TN) removal is limited due to unsatisfactory denitrification. This study demonstrated that short-time (1 h) pre-anoxic electro-stimulation significantly enhanced SND efficiency in the aerobic phase by promoting the proliferation of mixotrophic and heterotrophic denitrifiers. SND and TN removal efficiencies at the optimal electric current (EC) (0.02 A) were 85.6 % and 93.9 %, which were 39.1 % and 17.2 % higher than control. Microbial community analysis indicated that the abundance of mixotrophic and heterotrophic denitrifiers significantly increased. H 2 generated in the electro-stimulation process induced the proliferation of mixotrophic denitrifiers. The weak EC (0.02 A) promoted the activity and growth of heterotrophic denitrifiers by accelerating electron transfer. They concurrently mediated heterotrophic denitrification to enhance SND efficiency. PICRUSt2 analysis revealed that the abundance of denitrifying genes dramatically surged. This study provides new insights into applying electrolysis to achieve advanced SND while minimizing electricity consumption., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

An integrated process of sludge in-situ fermentation, biological phosphorus removal and endogenous denitrification (ISFPR-ED) was developed to treat low ratio of chemical oxygen demand to nitrogen (COD/N) wastewater and waste activated sludge (WAS) in a single reactor. Nutrient removal and WAS reduction were achieved due to Tetrasphaera-dominated sludge fermentation provided organic carbon in extending the anaerobic duration. The WAS reduction efficiency, effluent orthophosphate (PO 4 3- -P) and total inorganic nitrogen reached 28.1 %, less than 0.4 and 7.2 mg/L, respectively. While organic carbon was reduced by 67 %. Tetrasphaera, conventional polyphosphate accumulating organisms (PAOs) stored glycogen, amino acids, and PHA for nutrient removal. Excess energy from fermentation enhanced anaerobic PO 4 3- -P uptake by Tetrasphaera. Tetrasphaera was the dominant PO 4 3- -P removal and fermentation bacteria, working synergistically with conventional PAOs and fermenting microorganisms. This integrated process improves nutrient removal efficiency and reduces operating costs for carbon addition and WAS disposal in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Anaerobic ammonia oxidation (Anammox) exhibits promise for wastewater treatment,but the enrichment of anammox bacteria (AnAOB) in municipal wastewater treatment plants is a significant challenge. This study constructed a novel Anoxic-Anaerobic-Oxic (AAnO) process with a pure biofilm anoxic zone fed with actual fluctuating municipal wastewater and operated for six months to enrich AnAOB at ambient temperature. High-throughput sequencing (HTS), qPCR, and fluorescence in situ hybridization showed that AnAOB were successfully enriched in the anoxic biofilms, reaching 1.56 % relative abundance on day 75 detected by HTS. During the period from day 130 to day 186, the anammox process contributed to 55.8 ± 19.2 % of the nitrogen removal in the anoxic zone. Phylogenetic analysis revealed this AnAOB species was closely related to Candidatus Brocadia fulgida. This study provides technical support for the application of anammox in mainstream wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the rapid start-up of mainstream partial denitrification coupled with anammox (PD/A) and nitrogen removal performance by inoculating precultured PD/A biofilm. The results showed mainstream PD/A in the anaerobic-anoxic-aerobic (A 2 O) process was rapidly established within 30 days. Nitrogen removal efficiency (NRE) improved by 23.8 % contrasted to the traditional A 2 O process. The mass balance showed that anammox contribution to total nitrogen (TN) removal were maintained at 37.9 %∼55.7 %, and reducing hydraulic retention time (HRT) strengthened simultaneously denitrification and anammox activity. The microbial community showed that the dominant bacteria such as denitrifying bacteria (DNBs) and glycogen accumulating organisms (GAOs) both in biofilm and flocculent sludge (floc), integrating with anammox bacteria (AnAOB) in biofilm might lead to enhanced nitrogen removal. Overall, this study offered a fast start-up strategy of mainstream PD/A with enhanced nitrogen removal, which are valuable for upgradation and renovation of existed municipal wastewater treatment plants (WWTPs)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Manganese(IV) (Mn(IV)) reduction coupled to anaerobic ammonium (NH 4 + -N) oxidation (Mnammox) is a recently identified metal oxide-mediated nitrogen (N) loss pathway, holding potential value for the efficient removal of NH 4 + -N from wastewater. However, little is known about the application of Mnammox in wastewater treatment. Here, a novel Mnammox bacterium Aromatoleum evansii (strain MAY27) was screened. Strain MAY27 can utilize MnO 2 as an electron acceptor to achieve NH 4 + -N removal under a low C/N condition (C/N = 0.5). The influencing factors in the Mnammox process and the Mn(IV) reduction driving effect on NH 4 + -N oxidation were investigated. The physiological characteristics of strain MAY27 and differential metabolic pathways were identified through whole-genome sequencing and metabolomic analyses. A significant up-regulation of several key pathways upon the addition of MnO 2 , including glycolysis/gluconeogenesis, transmembrane transporter activity, and oxidoreductase activity. This study contributes to the advancement of biotechnological approaches for treating N-containing wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Zero-valent iron acts as an indirect electron donor, supplying ferrous iron for the nitrate-dependent ferrous oxidation (NDFO) process. The addition of activated carbon (AC) increased the specific NDFO activity in situ and ex situ by 0.4 mg-N/(d·g VSS) and 2.2 mg-N/(d·g VSS), respectively, due to the enrichment of NDFO bacteria. Furthermore, AC reduced the nitrous oxide emission potential of the sludge, a mechanism that metagenomic analysis suggests may act as a cellular energy storage strategy. During a 196-day experiment, a total nitrogen removal efficiency of 53.7 % was achieved, which may be attributed to the upregulation of key genes involved in iron oxidation and denitrification. Based on these findings, a model involving pilin, 'nanowires,' and a cyc2/?→/(FoxE→FoxY)/?→cymA/Complex III/?-mediated pathway for extracellular electron uptake was proposed. Overall, this work provides a feasible strategy for enhancing the nitrogen removal performance of the ZVI-NDFO process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Overexpression of Dunaliella parva (D. parva) malic enzyme (ME) gene (DpME) significantly increased DpME expression and ME enzyme activity in transgenic D. parva. Nitrogen limitation had an inhibitory effect on protein content, and DpME overexpression could improve protein content. Nitrogen limitation increased carbohydrate content, and Dunaliella parva overexpressing malic enzyme gene under nitrogen limitation (DpME-N - ) group showed the lowest starch content among all groups. Dunaliella parva overexpressing malic enzyme gene under nitrogen sufficient condition (DpME) and DpME-N - groups showed considerably high mRNA levels of DpME. ME activity was significantly enhanced by DpME overexpression, and nitrogen limitation caused a smaller increase. DpME overexpression and nitrogen limitation obviously enhanced lipid accumulation, and DpME overexpression had more obvious effect. Compared with control (wild type), lipid content (68.97%) obviously increased in DpME-N - group. This study indicated that the combination of DpME overexpression and nitrogen limitation was favorable to the production of microalgae biodiesel., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study proposed the double-edged sword effects of sulfate reduction process on nitrogen removal and antibiotic resistance genes (ARGs) transmission in sulfur autotrophic denitrification system. Excitation-emission matrix-parallel factor analysis identified the protein-like fraction in soluble microbial products as main endogenous organic matter driving the sulfate reduction process. The resultant sulfide tended to serve as bacterial modulators, augmenting electron transfer processes and mitigating oxidative stress, thereby enhancing sulfur oxidizing bacteria (SOB) activity, rather than extra electron donors. The cooperation between SOB and heterotroph (sulfate reducing bacteria (SRB) and heterotrophic denitrification bacteria (HDB)) were responsible for advanced nitrogen removal, facilitated by multiple metabolic pathways including denitrification, sulfur oxidation, and sulfate reduction. However, SRB and HDB were potential ARGs hosts and assimilatory sulfate reduction pathway positively contributed to ARGs spread. Overall, the sulfate reduction process in sulfur autotrophic denitrification system boosted nitrogen removal process, but also increased the risk of ARGs transmission., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study used 16S rRNA gene sequencing and metatranscriptomic analysis to comprehensively illustrate how ammonia stress influenced medium-chain fatty acids (MCFA) biosynthesis. MCFA synthesis was inhibited at total ammonia nitrogen (TAN) concentrations above 1000 mg N/L. TAN stress hindered organic hydrolysis, acidification, and volatile fatty acids elongation. Chain-elongating bacteria (e.g., Clostridium_sensu_stricto_12, Clostridium_sensu_stricto_1, Caproiciproducens) abundance remained unchanged, but their activity decreased, partially due to the increased reactive oxygen species. Metatranscriptomic analysis revealed reduced activity of enzymes critical for MCFA production under TAN stress. Fatty acid biosynthesis pathway rather than reverse β-oxidation pathway primarily contributed to MCFA production, and was inhibited under TAN stress. Functional populations likely survived TAN stress through osmoprotectant generation and potassium uptake regulation to maintain osmotic pressure, with NADH-ubiquinone oxidoreductase potentially compensating for ATP loss. This study enhances understanding of MCFA biosynthesis under TAN stress, aiding MCFA production system stability and efficiency improvement., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The combination of microalgal culture and wastewater treatment is an emerging topic. This study investigated the use of different microalgae to treat different types of dairy farm wastewater. The results showed that the removal of ammonia nitrogen and total phosphorus by mixed microalgae was over 99% and 80%, respectively. The highest production of protein in biomass and extracellular polymeric substances was observed in high-concentration wastewater. In the phycosphere, the abundance of Proteobacteria and Cyanobacteria increased, while that of Bacteroidota decreased. Phycosphere bacteria were strongly correlated with microalgal growth and the composition of extracellular polymeric substances, especially with bound extracellular polymeric substances relative to soluble extracellular polymeric substances. Genes associated with photosynthesis and respiration in phycosphere bacteria were upregulated, contributing to the material exchange capacity in the microalgal-bacterial systems. The interaction between microalgae and phycosphere bacteria thus represents the core of the binary cultivation system-based wastewater treatment and requires further investigation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Heterotrophic nitrification (HN) bacteria use organic carbon sources to remove ammonia nitrogen (NH 4 -N); however, the mechanisms of carbon and nitrogen metabolism are unknown. To understand this mechanism, HN functional microbial communities named MG and MA were enriched with glucose and sodium acetate, respectively. The NH + -N); however, the mechanisms of carbon and nitrogen metabolism are unknown. To understand this mechanism, HN functional microbial communities named MG and MA were enriched with glucose and sodium acetate, respectively. The NH 4 + -N removal efficiencies were 98.87 % and 98.91 %, with 88.06 % and 69.77 % nitrogen assimilation for MG and MA at 22 h and 10 h, respectively. Fungi (52.86 %) were more competitive in MG, and bacteria (99.99 %) were dominant in MA. Metagenomic and metabolomic analyses indicated that HN might be a signaling molecule (NO) in the production and detoxification processes when MG metabolizes glucose (amo, hao, and nosZ were not detected). MA metabolizes sodium acetate to produce less energy and promotes nitrogen oxidation reduction; however, genes (hao, hox, and NOS2) were not detected. These results suggest that NO and energy requirements induce microbial HN., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A novel A. pittii J08 with heterotrophic nitrification and aerobic denitrification (HN-AD) isolated from pond sediments could rapidly degrade inorganic nitrogen (N) and total nitrogen (TN-N) with ammonium (NH 4 + -N) preference. N degradation rate of NH 4 + -N, nitrite (NO 2 - -N) and nitrate (NO 3 - -N) were 3.9 mgL -1 h -1 , 3.0 mgL -1 h -1 and 2.7 mgL -1 h -1 , respectively. In addition, strain J08 could effectively utilize most of detected low-molecular-weight carbon (LMWC) sources to degrade inorganic N with a wide adaptability to various culture conditions. Whole genome sequencing (WGS) analysis revealed that assembled genome of stain J08 possessed the crucial genes involved in dissimilatory/assimilatory NO 3 - -N reduction and NH 4 + -N assimilation. These results indicated that strain J08 could be applied to wastewater treatment in aquaculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The extremely slow growth rate of anaerobic ammonia oxidation (anammox) bacteria limits full-scale application of anammox process worldwide. In this study, extracellular polymeric substances (EPS)-coated polypropylene (PP) carriers were prepared for biofilm formation. The biomass adhesion rate of EPS-PP carrier was 12 times that of PP carrier, and EPS-PP achieved significant enrichment of E. coli BY63. The 120-day continuous flow experiment showed that the EPS-PP carrier accelerated the formation of anammox biofilm, and the nitrogen removal efficiency increased by 10.5 %. In addition, the abundance of Candidatus Kuenenia in EPS-PP biofilm was 27.1%. Simultaneously, amino acids with high synthesis cost and the metabolites of glycerophospholipids related to biofilm formation on EPS-PP biofilm were significantly up-regulated. Therefore, EPS-PP carriers facilitated the rapid formation of anammox biofilm and promoted the metabolic activity of functional bacteria, which further contributed to the environmental and economic sustainability of anammox process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This research evaluated a microalgae consortium (MC) in a pilot-scale tubular photobioreactor for municipal wastewater (MWW) treatment, compared with an aeration column photobioreactor. Transitioning from suspended MC to a microalgae-microbial biofilm (MMBF) maintained treatment performance despite increasing influent from 50 L to 150 L in a 260 L system. Carbon and nitrogen removal were effective, but phosphorus removal varied due to biofilm shading and the absence of phosphorus-accumulating organisms. High influent flow caused MMBF detachment due to shear stress. Stabilizing and re-establishing the MMBF showed that a stable phycosphere influenced microbial diversity and interactions, potentially destabilizing the MMBF. Heterotrophic nitrification-aerobic denitrification bacteria were crucial for MC equilibrium. Elevated gene expression related to nitrogen fixation, organic nitrogen metabolism, and nitrate reduction confirmed strong microalgal symbiosis, highlighting MMBF's treatment potential. This study supports the practical application of microalgae in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Sponge iron (SFe) coupled with a sludge system has great potential for improving biological denitrification; however, the underlying mechanism is not yet fully understood. In this study, the denitrification performance and microbial characteristics of ordinary sludge and SFe-sludge systems were investigated. Overall, the SFe-sludge reactor had faster ammonium degradation rate (94.0 %) and less nitrate accumulation (1.5-53.3 times lower) than ordinary reactor during the complete operation cycle of sequencing batch reactors. The addition of SFe increased the activities of nitrate and nitrite reductases. The total relative abundance of autotrophic denitrifying bacteria (Acidovorax, Arenimonas, etc.) in the SFe-sludge system after 38 days of operation was found to be 10.6 % higher than that in the ordinary sludge reactor. The aerobic denitrifying bacteria (Dokdonella, Phaeodactylibacter, etc.) was 5.3 % higher than ordinary sludge. The SFe-sludge system improved denitrification by enriching autotrophic/aerobic denitrifying bacteria in low carbon-to-nitrogen ratio wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Immobilized microalgae biotechnologies can conserve water and space by low-carbon wastewater treatment and resource recovery in a recirculating aquaculture system (RAS). However, technical process parameters have been unoptimized considering the mutual interaction between factors. In this study, machine learning optimized the parameters of alginate-immobilized Chlorella vulgaris (C. vulgaris), that is, 474 μmol/(m 2 ·s) of light intensity, 23 × 10 6 cells/mL for initial cell number, and 2.07 mm particle size. Importantly, under continuous illumination, the immobilized C. vulgaris and microalgal-bacterial consortium improved water purification and biomass reutilization. Transcriptomics of C. vulgaris showed enhanced nitrogen removal by increasing pyridine nucleotide and lipid accumulation via enhanced triacylglycerol synthesis. Symbiotic bacteria upregulated genes for nitrate reduction and organic matter degradation, which stimulated biomass accumulation through CO 2 fixation and starch synthesis. The recoverable microalgae (1.94 g/L biomass, 47 % protein, 26.23 % lipids), struvite (64.79 % phosphorus), and alginate (79.52 %) every two weeks demonstrates a low-carbon resource recovery in RAS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Anammox is recognized as a prospective alternative for future biological nitrogen removal technologies. However, the nitrate by-products produced by anammox bacteria limit its overall nitrogen removal efficiency below 88 %. This study introduced Fe(III) into the anammox bioreactor to enhance the nitrogen removal efficiency to approximately 95 %, surpassing the biochemical limit of 88 % imposed by anammox stoichiometry. Anammox sludge was demonstrated to utilize extracellular polymeric substances to reduce Fe(III) into Fe(II), and this process promoted the dominance of Ca. Brocadia. The iron addition improved the abundance of narGHI genes and facilitated the partial dissimilatory nitrate reduction to ammonium, with nitrite as the end product. The accumulated nitrite was then eliminated through the anammox pathway, along with the excess ammonium (30 mg/L) in the influent. Overall, this study deepens our understanding of the enhanced nitrogen removal triggered by Fe(III) in anammox sludge and offers an effective approach to boost anammox process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Rare earth elements result in substantial tailings wastewater with high ammonium and nitrate during extraction. In this study, a temperature-resilient Anammox process was employed for efficient treatment of rare earth element tailings wastewater through implementing synergistic nitrite supply by partial nitritation (PN) and partial denitrification (PD). Enhancing temperature resilience of Anammox process relies on dynamic management of DO and COD inputs to shift the dominant nitrite supplier from PN to PD, stable PD (NAR ≥ 90 %) can boost nitrogen removal by Anammox to 97.8 %. The nitrogen removal rate and nitrogen removal efficiency at 10.6 °C could maintain at 0.12 kgN/m 3 ·d -1 and 92.5 %, respectively. Microbial analysis reveals that Nitrosomonas, Thauera, and Candidatus_Kuenenia are the predominant genera responsible for nitrite supply and nitrogen removal, localized within the gas channels of granules, flocs, and micro-granules, respectively. Keeping the influent C/NO 3 - -N ratio below 1.7 is ideal to prevent overgrowth of Thauera and maintain system stability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The phytopromotional root endophytic fungus Piriformospora indica was introduced into the wetland plant Canna indica L. to explore its impact on nitrogen (N) removal in constructed wetlands (CWs) to treat normal and saline (0.9 % NaCl) wastewater. P. indica colonization increased total nitrogen, NH 4 + -N, and NO 3 - -N removal efficiencies under normal and saline conditions, with NO 3 - -N removal rates significantly increasing by 17.5 % under saline conditions (P<0.05). N removal by plant uptake improved by 26.1 % and 27.7 % under normal and saline conditions due to P. indica-mediated growth-promoting effects. Salt-tolerant denitrifiers and nitrifiers guaranteed the dominant role of microbial degradation in N removal under saline conditions. P. indica inoculation considerably improved the contribution of Nocardioides and Nitrosomnas to dissimilatory/assimilatory nitrate reduction and nitrification genes, respectively. These findings elucidate the mechanisms and potential applications of P. indica-mediated phytoremediation in practical wastewater treatment under varying salty conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

[Display omitted] • Iron-carbon (IC) enhances simultaneous nitrification and denitrification of AGS. • Nitrogen metabolism variation was analyzed by metagenomics and enzyme activity. • IC reduces nitrite accumulation and nitrous oxide production potential from AGS. • IC acts as an activator of enzymatic reactions. The aerobic granular sludge (AGS) process had been extensively studied for its simultaneous nitrification and denitrification (SND) capabilities. Iron-carbon (IC) had enhanced AGS nitrogen removal efficiency, but the mechanism remained unclear. In this study, four reactors had been added with 50, 30, 10, and 0 g/L of IC. Total nitrogen removal efficiency increased with IC dosage under the same operation mode. IC enhanced sludge ammonia oxidation rate, denitrification rate, and specific oxygen uptake rate, allowing SND to complete 60 min earlier, potentially reducing wastewater treatment costs. Notably, IC eliminated nitrite accumulation in conventional AGS effluent. IC decreased the abundance of genes and enzyme activities related to NOR expression, while increasing those related to NOS, which may mitigate the potential for nitrous oxide formation by microorganisms. In this study, IC acted as an enzymatic reaction activator, affecting granules more than flocs, with the activity gap gradually decreasing with the IC dosage. [ABSTRACT FROM AUTHOR]

[Display omitted] • Higher nitrogen removal was achieved in FeS 2 -based CWs under light conditions. • Light conditions increased organic carbon input into the rhizosphere. • Higher TOC content promoted the enrichment of denitrifiers. • Iron cycle and glycolysis supply more e− for denitrification in higher TOC content. In constructed wetlands (CWs), carbon source availability profoundly affected microbial metabolic activities engaged in both iron cycle and nitrogen metabolism. However, research gaps existed in understanding the biotransformation of nitrogen and iron in response to fluctuations in organic carbon content under day-night alterations. Results demonstrated increased removal efficiency of NO 3 −-N (95.7 %) and NH 4 +-N (75.70 %) under light conditions, attributed to increased total organic carbon (TOC). This enhancement promoted the relative abundance of bacteria involved in nitrogen and iron processes, establishing a more stable microbial network. Elevated TOC content also upregulated genes for iron metabolism and glycolysis, facilitating denitrification. Spearman correlation analysis supported the synergistic mechanisms between FeS 2 -based autotrophic denitrification and TOC-mediated heterotrophic denitrification under light conditions. The significant impact of carbon sources on microbial activities underscores the critical role of organic carbon availability in enhancing nitrogen removal efficiency, providing valuable insights for optimizing FeS 2 -based CWs design and operation strategies. [ABSTRACT FROM AUTHOR]

[Display omitted] • Microalgal-bacterial granular sludge-marimo (MBGS-MA) process was established. • The MBGS-MA coupling process could realize efficient nitrogen (N) removal. • The assimilatory nitrate reduction and assimilation in MA were enhanced. • Ammonia starvation promoted enrichment of N fixing genes (nifDKH) in MA. Current biological wastewater treatment processes usually have a drawback of insufficient nitrogen (N) removal, contributing to the ubiquitous eutrophication of aquatic ecosystems globally. To address such a challenging situation, this study explored an innovative microalgal-bacterial granular sludge-marimo (MBGS-MA) coupling process. The process removed 83.4 % of N with the effluent N concentration of 4.0 mg/L. With the growth of MBGS, there was a shift towards genes associated with nitrification and denitrification, and away from ammonia assimilation genes, revealing internal mechanism of the shift of N removal pathway. Contrarily, MA could use gaseous N 2 with the N fixing genes in MA enriched, and the genes abundance related to assimilatory nitrate reduction were also raised under the mutualistic interactions between Proteobacteria and Cyanobacteria , which was beneficial to achieve efficient N removal. These findings may open a new horizon for developing innovative hybrid microalgal-bacterial processes aimed at high-efficiency N removal from wastewater. [ABSTRACT FROM AUTHOR]

Low temperature has always been a significant limitation for the biological removal of ammonia nitrogen (NH 3 -N) from water. Acinetobacter harbinensis HITLi7 T (HITLi7 T ) was used to enhance the low-temperature nitrification biofilter (LTNB) with a treatment capacity of 20,000 m 3 /d. At 2 °C, with an empty bed contact time of 3 h, the LTNB achieved NH 3 -N removal levels of 1.2 ∼ 1.5 mg/L. The nitrifying bacteria (Nitrosomonas, Nitrosospira, Nitrospira and Candidatus&#95;Nitrotoga) were significantly enriched. PICRUSt2 and FAPROTAX revealed the nitrification pathway of NH 3 -N conversion to hydroxylamine, then to nitrite, and finally to nitrate. The high co-occurrence of HITLi7 T with the nitrifying bacteria suggested that HITLi7 T might also promote the enrichment of nitrifying bacteria. Life cycle assessment showed that LTNB was an economical and environmentally friendly method for NH 3 -N removal. These results indicated that HITLi7 T enhanced the nitrification performance of biofilters, improved the cold tolerance of nitrifying bacteria, and had potential for practical applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This work unraveled discrepant ecological patterns between biofilms and flocs in a deteriorated bioreactor inoculated with mature partial nitrification-anammox (PN/A) sludge. Based on 16S rRNA analysis, a comprehensive evaluation of neutral and null models, along with niche width, delineated that the bacterial community assembly in biofilms and flocs was dominantly driven by the stochastic process, and dispersal limitation critically shaped the community assembly. Co-occurrence network analysis revealed that environmental stress caused decentralized and fragmented bacterial colonies, and anammox bacteria were mainly peripheral in biofilms network and less involved in interspecific interactions. Simultaneous PN/A and partial denitrification-anammox (PD/A) processes were identified, whereas PN and PD process primarily occurred in the biofilms and flocs, respectively, as evidenced by metagenomics. Collectively, these outcomes are expected to deepen the basic understanding of complex microbial community and nitrogen metabolism under environmental disturbance, thereby better characterizing and serving the artificial ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Magnesium ion (Mg 2+ ) plays an important role in the accumulation and stability of anaerobic ammonium-oxidizing bacteria (AnAOB). In this study, the response of anammox sludge to Mg 2+ was comprehensively investigated by performance evaluation and metagenomics analysis. Appropriate Mg 2+ (0.8 mmol/L) could improve the nitrogen removal performance, AnAOB activity, and the synthesis potential of some hydrophobic substances, while high Mg 2+ (>1.6 mmol/L) has a negative effect. Meanwhile, Mg 2+ transmembrane transport theory was introduced to reveal the response principle of AnAOB to Mg 2+ from a novel insight. AnAOB may have a self-defense function based on the PhoQ/PhoP-MgtAB system. Low extracellular Mg 2+ will activate this function to enhance Mg 2+ influx, thereby improving the intracellular metabolism of AnAOB. Excessive Mg 2+ , however, dormant this function and induces Mg 2+ efflux, which may decrease the intracellular Mg 2+ and thus affect AnAOB metabolism. These findings provide valuable references for the Mg 2+ regulation of anammox-based process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Sulfur-mediated autotrophic biological nitrogen removal (BNR) processes favor the reduction of greenhouse gas (GHG) emissions compared to heterotrophic BNR processes. Micro-oxygen environments are widely prevalent in practical BNR systems, and the mechanisms of GHG emissions mediated by multi-elements, including nitrogen (N), sulfur (S), and oxygen (O), remain to be systematically summarized. This review reveals the functional microorganisms involved in sulfur-mediated BNR processes under micro-oxygen regulation, elucidating their metabolic mechanisms and interactions. The GHG abatement potential of sulfur-mediated BNR processes under micro-oxygen regulation is highlighted, along with recent advances in multi-scenario applications. The fate of GHG in wastewater treatment systems is explored and insights into future multi-scale GHG regulatory strategies are provided. Overall, the application of sulfur-mediated BNR processes under micro-oxygen regulation exhibits great potential. This review can act as a guide for the effective implementation of strategies to mitigate the environmental impacts of GHG emissions from wastewater treatment processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Efficient nitrogen removal yeasts are rarely encountered. Here, a heterotrophic nitrification-aerobic denitrification strain of Candida boidinii L21 was isolated. The optimal removal conditions for strain L21 were glucose as carbon source, C/N of 15, salinity of 10 ppt, pH of 7, shaking speed of 120 rpm, and temperature of 30 °C. Strain L21 removed NH 4 + -N, NO 2 - -N, NO 3 - -N (14---140 mg/L) and achieved nearly complete NO 2 - -N, removal. Nitrogen balance and enzyme activity analysis indicated the nitrogen removal pathway of strain L21 through assimilation, nitrification, and denitrification pathways. When applied in wastewater and sludge, strain L21 reduced inorganic nitrogen levels within 4 days, with a 58-fold increase in nitrite removal compared to controls. These findings demonstrate that strain L21 holds great potential for enhancing nitrogen removal in wastewater treatment processes, providing valuable insights for improving environmental management practices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cupric ions can restrain biological nitrogen removal processes, which comprise nitrite reductase and nitric oxide reductase. Here, Pseudomonas sp. NY1 can efficiently perform heterotrophic nitrification and aerobic denitrification with cupric ions at 15 °C. At optimal culturing conditions, low cupric ion levels accelerated nitrogen degradation, and ammonium and nitrite removal efficiencies increased by 2.33%-4.85% and 6.76%-12.30%, respectively. Moreover, the maximum elimination rates for ammonium and nitrite increased from 9.48 to 10.26 mg/L/h and 6.20 to 6.80 mg/L/h upon adding 0.05 mg/L cupric ions. Additionally, low cupric ion concentrations promoted electron transport system activity (ETSA), especially for nitrite reduction. However, high concentrations of cupric ions decreased the ETSA during nitrogen conversion processes. The crucial enzymes ammonia monooxygenase, nitrate reductase, and nitrite reductase possessed similarly trends as ETSA upon exposure to cupric ion. These findings deepen the understanding for the effect of cupric ions on nitrogen consumption and bioremediation in nitrogen-polluted waters., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: This work was financially supported by the National Natural Science Foundation of China (No. 42,167,019 and 42007223). Scientific Research Inovation Team Project of Guizhou University [No. Guidakechuangtuan (2024) 06]., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The partial nitritation-anammox process based on a membrane-aerated biofilm reactor (MABR) faces several challenges, such as difficulty in suppressing nitrite-oxidizing bacteria (NOB), excessive effluent nitrate, and ineffective synergy between denitrification and anammox bacteria. Therefore, a novel partitioned granular sludge coupling with MABR (G-MABR) was constructed. The chemical oxygen demand (COD) and nitrogen removal efficiency were 88.8 ± 1.8 %-92.6 ± 1.2 % and 88.8 ± 1.5 %-93.6 ± 0.7 %, respectively. The COD was mainly lowered in the lower granular sludge-zone, while nitrogen was removed in the upper MABR-zone. NOB was significantly suppressed in the MABR-zone due to competition for substrate with denitrifying bacteria and anammox bacteria. This partitioned configuration reduced the C/N ratio in the MABR-zone, thus facilitating autotrophic nitrogen removal. Both partial nitrification and denitrification provided nitrite for anammox bacteria in granular sludge, whereas partial nitrification mainly supplied nitrite to the anammox bacteria in membrane biofilms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Anammox-based processes are pivotal for elevating nitrogen removal efficiency in municipal wastewater treatment. This study established a novel HF-EPDA system combined in-situ hydrolytic fermentation (HF) with endogenous partial denitrification (EPD) and anammox. Slowly-biodegradable organic matter (SBOM) was degraded and transformed into endogenous polymers for driving production of sufficient nitrite by EPD, further promoted the nitrogen removal via anammox process. Processes above formed positive feedback, guaranteeing the robustness and recoverability of system. After a 92-day suspension during operation, advanced nitrogen removal was still achieved with excellent nitrogen removal efficiency of 95.84 ± 1.73 %, treating with actual domestic wastewater and synthetic nitrate wastewater. Candidatus Brocadia and Candidatus Competibacter were dominant bacteria on biofilms responsible for the anammox and EPD process respectively, while the main hydrolytic fermentation organisms norank&#95;o SBR1031 was enriched in floc sludge. This study highlights the reliable potential for expanding anammox application with simultaneous improvement of SBOM utilization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Excessive nitrogen release during composting poses significant challenges to both the environment and compost quality. Biological enhancement of humification and nitrogen conservation is an environmentally friendly and cost-effective approach to composting. The aim of this study was to develop a psychrophilic and thermophilic nitrifying bacterial consortium (CNB) and investigate its role in nitrogen transformation and humification during cow manure composting. Analysis revealed that CNB inoculation promoted microbial proliferation and metabolism, significantly increased the number of nitrifying bacteria (p < 0.05), and elevated the activity of nitrite oxidoreductase and nxrA gene abundance. Compared to the control, CNB inoculation promoted the formation of NO 3 - -N (77.87-82.35 %), while reducing NH 3 (48.89 %) and N 2 O (20.05 %) emissions, and increased humus content (16.22 %). Mantel analysis showed that the higher abundance of nitrifying bacteria and nxrA facilitated the nitrification of NH 4 + -N. The improvement in nitrite oxidoreductase activity promoted NO 3 - -N formation, leading to increased humus content and enhanced compost safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Overcoming low nitrogen removal efficiency at low temperatures is a challenge in biological treatment. This study investigated the cold-tolerant heterotrophic nitrification-aerobic denitrification by Acinetobacter calcoaceticus TY1. Transcriptomic and biochemical analyses indicated that strain TY1 upregulated genes for energy production, assimilation, cell motility, and antioxidant enzyme production under cold stress, maintaining functions such as energy supply, nitrogen utilization, and oxidative defense. Increasing the synthesis of extracellular polysaccharides, unsaturated fatty acids, and medium-chain fatty acids and secreting large amounts of antioxidant enzymes ensured cell membrane flexibility while enhancing the antioxidant system. Immobilization experiments showed that biofilms accelerated the removal of nitrogen pollutants and demonstrated good stability, with carriers being reusable to five times, maintaining high ammonia nitrogen (63.90 %) and total nitrogen (50.66 %) removal rates. These findings reveal the cold tolerance mechanisms of strain TY1 and its excellent practical potential as a candidate for wastewater treatment in cold regions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Two parallel pilot-scale reactors were operated to investigate pollutant removal performance and metabolic pathways in elemental sulfur-driven autotrophic denitrification (SDAD) process under low temperature and after addition of external electron donors. The results showed that low temperature slightly inhibited SDAD (average total nitrogen removal of ∼4.7 mg L -1 ) while supplement of sodium thiosulfate (stage 2) and sodium acetate (stage 3) enhanced denitrification and secretion of extracellular polymeric substances (EPS), leading to the average removal rate of 0.75 and 1.01 kg N m -3  d -1 , respectively with over twice higher total EPS. Correspondingly, nitrogen and sulfur related microbial metabolisms especially nitrite reductase and nitric oxide reductase encoding were promoted by genera including Thermomonas and Thiobacillus. The variations revealed that extra sodium acetate improved denitrification and enriched more SDAD-related microorganisms compared with sodium thiosulfate, which potentially catalyzed the refinement of practical strategies for optimizing denitrification in low carbon to nitrogen ratio wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigates the performance, resilience and microbial community dynamics of two anaerobic processes, i.e. pure anammox (R1) and partial denitrification/anammox (PD/A) (R2), following a 30-day starvation period. The tolerance to starvation was assessed by comparing nitrogen removal efficiency and microbial activity across both reactors. Results show that the PD/A process recovery to pre-starvation performance levels within just one day, as compared to the pure anammox process. Notably, although the activity of anammox bacteria decreased in both processes during starvation, the decay rate in R1 was 69.59 % higher than in R2, potentially explaining the quicker recovery of R2. Furthermore, enhanced secretion of extracellular polymeric substance (EPS) during starvation served as a protective mechanism. The potential functions and genes in microorganisms, as well as the pathway of nitrogen cycling, were demonstrated through analyses using the KEGG database. This research reveals essential mechanistic insights and strategic guidance for the effective implementation of anammox-based biological nitrogen removal processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The feasibility of inducing simultaneous nitrification and denitrification (SND) by S 0 for low carbon to nitrogen (C/N) ratio wastewater remediation was investigated. Compared with S 0 and/or organics absent systems (-3.4 %∼5.0 %), the higher nitrogen removal performance (18.2 %∼59.8 %) was achieved with C/N ratios and S 0 dosages increasing when S 0 and organics added simultaneously. The synergistic effect of S 0 and organics stimulated extracellular polymeric substances secretion and weakened intermolecular binding force of S 0 , facilitating S 0 bio-utilization and reducing the external organics requirement. It also promoted microbial metabolism (0.16 ∼ 0.24 μg O 2 /(g VSS·h)) and ammonia assimilation (5.9 %∼20.5 %), thereby enhancing the capture of organics and providing more electron donors for SND. Furthermore, aerobic denitrifiers (15.91 %∼27.45 %) and aerobic denitrifying (napA and nirS) and ammonia assimilating genes were accumulated by this synergistic effect. This study revealed the mechanism of SND induced by coordination of S 0 and organics and provided an innovative strategy for triggering efficient and stable SND., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

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 PO 4 3 - -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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The nitrogen loss in composting is primarily driven by the transformation of organic nitrogen, yet the mechanisms underlying the degradation process remain incompletely understood. This study employed protein family domains (Pfams) analysis based on metagenomic sequencing to investigate the functional characteristics, key microorganisms, and environmental parameters influencing organic nitrogen degradation in chicken manure and pig manure composting. 154 Pfams associated with ammonification function were identified. Predominant Pfams: proteolytic peptidase, followed by chitin/cell wall degraders, least involved in nucleic acid degradation. Ammonifying microbial diversity was basically consistent among compost types, particularly in the thermophilic stage with the peak of abundance of dominant ammonifying microorganisms. Viruses played an important role in ammonification process, especially Uroviricota. 26 key ammonifying genera were identified by the microbial network. pH dominated the metabolic activity of ammonifying microorganisms in various manure compost types, primarily consisting of protein-degrading bacteria with stable community structures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cupriavidus necator is a facultative chemolithoautotrophic bacterium able to convert carbon dioxide into poly-3-hydroxybutyrate. This is highly promising as the conversion process allows the production of sustainable and biodegradable plastics. Poly-3-hydroxybutyrate accumulation is known to be induced by nutrient starvation, but information regarding the optimal stress conditions controlling the process is still heterogeneous and fragmentary. This study presents a comprehensive comparison of the effects of nutrient stress conditions, namely nitrogen, hydrogen, phosphorus, oxygen, and magnesium deprivation, on poly-3-hydroxybutyrate accumulation in C. necator DSM545. Nitrogen starvation exhibited the highest poly-3-hydroxybutyrate accumulation, achieving 54% of total cell dry weight after four days of nutrient stress, and a carbon conversion efficiency of 85%. The gas consumption patterns indicated flexible physiological mechanisms underlying polymer accumulation and depolymerization. These findings provide insights into strategies for efficient carbon conversion into bioplastics, and highlight the key role of C. necator for future industrial-scale applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

To reveal the key enzymes in the nitrogen removal pathway and to further elucidate the mechanism of the catalytic reaction, this study utilized metaproteomics combined with molecular dynamics and density functional theory calculation. K. stuttgartiensis provided the proteins up to 88.37 % in the anammox-based system. Hydrazine synthase (HZS) and hydrazine dehydrogenase (HDH) accounted for 15.94 % and 3.45 % of the total proteins expressed by K. stuttgartiensis, thus were considered as critical enzymes in the nitrogen removal pathway. The process of HZSγ binding to NO with lowest binding free energy of -4.91 ± 1.33 kJ/mol. The reaction catalyzed by HZSα was calculated to be the rate-limiting catalyzing step, because it transferred the proton from NH 3 to ·OH by crossing an energy barrier of up to 190.29 kJ/mol. This study provided molecular level insights to enhance the performance of nitrogen removal in anammox-based system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study addressed the treatment of high ammonia, low biodegradable chemical oxygen demand (bCOD) anaerobically digested molasses wastewater, utilizing an aerobic granular sludge (AGS) reactor. The AGS achieved 99 % ammonia removal regardless of the bCOD supplementation. By adding low ammonia (<60 mg/L), high bCOD raw molasses wastewater (before anaerobic digestion) as a carbon source, enhanced nitrogen removal, increasing from 10 % to 97 %, and improved sludge settleability via bio-induced calcite precipitation were observed. Functional genes prediction suggested two potential denitrification pathways, including heterotrophic denitrification by Paracoccus and Thauera, and autotrophic denitrification, specifically sulfide-oxidizing autotrophic denitrification by Thiobacillus. An increase in the relative abundance of microorganisms involved in heterotrophic denitrification was observed with the addition of high bCOD raw molasses wastewater. Consequently, incorporating raw molasses wastewater into the AGS presents a sustainable approach to achieve mixotrophic denitrification, maintain stable granular sludge and ensure stable treatment performance when treating anaerobically digested molasses wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Effluent from anammox granular sludge (AnGS) bioreactor contains microbes and microbial products. This study explored mechanisms of utilizing AnGS-effluent as biostimulant for anammox process enhancement. Compared with no AnGS-effluent supplemented control reactor, 5.0 and 1.3 times higher ammonium nitrogen and total inorganic nitrogen removal rates, respectively were obtained with continuous AnGS-effluent supplementation after 98 days' operation. Anammox bacteria from Candidatus Brocadia accounted for 0.1 % (DNA level) and 1.3 %-1.5 % (RNA level) in control reactor, and 2.9 % (DNA level) and 54.5 %-55.4 % (RNA level) in the AnGS-effluent-fed reactor. Influent microbial immigration evaluation showed that bacterial immigration via AnGS-effluent supplementation was not the main contributor to active anammox community development. Amino acids biosynthesis, B-vitamins and coenzymes metabolism related pathways were facilitated by AnGS-effluent supplementation. AnGS-effluent supplementation aided anammox metabolic activity by shaping microenvironment and microbial interactions. This study provides insights into enhancing anammox bacterial metabolism with AnGS-effluent microbial products as biostimulant., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, two bioprocess models were first constructed with the newly-discovered comammox process described as one-step and two-step nitrification and evaluated against relevant experimental data. The validated models were then applied to reveal the potential effect of comammox bacteria on the granular bioreactor particularly suitable for undertaking partial nitritation/anammox (PN/A) under different operating conditions of bulk dissolved oxygen (DO) and influent NH 4 + . The results showed although comammox bacteria-based PN/A could achieve > 80.0 % total nitrogen (TN) removal over a relatively wider range of bulk DO and influent NH 4 + (i.e., 0.25-0.40 g-O 2 /m 3 and 470-870 g-N/m 3 , respectively) without significant nitrous oxide (N 2 O) production (< 0.1 %), the bulk DO should be finely controlled based on the influent NH 4 + to avoid the undesired full nitrification by comammox bacteria. Comparatively, conventional ammonium-oxidizing bacteria (AOB)-based PN/A not only required higher bulk DO to achieve > 80.0 % TN removal but also suffered from 1.7 %∼2.8 % N 2 O production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Higher nitrite accumulation, which is challenging to achieve reliably, is always sought to obtain better nitrogen removal performance in traditional partial nitritation-anammox (PN/A) process. This study developed a modified PN/A process by introducing nitrite-oxidizing bacteria and endogenous metabolism. Advanced nitrogen removal performance of 95.5 % was achieved at a low C/N ratio of 2.7 under nitrite accumulation ratio (NAR) fluctuations. Higher nitrate accumulation at lower NAR (70 ∼ 40 %) resulted in superior anammox contribution (60 ∼ 75 %) and nitrogen removal performance (93 ∼ 98 %). This was attributed to the higher nitrogen removal efficiency of the post-anoxic endogenous partial denitrification coupling anammox process, although the PN/A process occurring first possessed a faster anammox rate of 2.0 mg NH 4 + -N /(g VSS⋅h). The introduction of nitrate allowed more nitrite flow to anammox, promoting a high enrichment of anammox bacteria (Ca. Brocadia, 0.3 % to 2.8 %). This study provides new insights into the practical application of the PN/A process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cyanobacteria hold promise for simultaneous carbon capture and chemicals production, but the regulation and effect of nitrogen (N) and phosphorus (P) remains unclear. This study investigates major productions of glycogen, protein, and C-phycocyanin (C-PC) in Cyanobacterium aponinum PCC10605 under different N/P levels, alongside changes in light and CO 2 . Increasing nitrate (NO 3 - ) from 2 to 6 mM resulted in a 9.7-fold increase in C-PC and reduced glycogen to 8.9 %. On the other hand, elevating phosphorus from 0.1 to 2 mM under limited nitrogen enhanced biomass and glycogen through the upregulation of carbonic anhydrase, ADP-glucose pyrophosphorylase, and glycogen phosphorylase. Changes in phosphorus levels and CO 2 inlet concentrations affected metabolites accumulation and carbon capture efficiency, leading to the best condition of 76 % uptake capacity in direct air capture (DAC). All findings underscore the trade-off between glycogen and protein, representing the importance of N/P levels in nutrient modulation of PCC10605., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study constructed an integrated algae/partial nitrification/anammox biofilm system and operated it for 240 days. The total nitrogen removal efficiency exceeded 90 %. The structure, compositions, and function of this symbiotic biofilm, which played a pivotal role in the system, were analyzed in detail. Microscope photos and fluorescence in situ hybridization both showed that bacteria and algae were well integrated. The dissolved oxygen gradient further confirmed that different functional microorganisms grew at varying depths within biofilm. Algae formed an oxygen-producing zone (0-0.48 mm), followed by ammonia oxidizing bacteria (AOB) consuming oxygen to form an oxygen-consuming zone (0.48-0.86 mm), and anaerobic ammonia oxidizing bacteria (AnAOB) removed nitrogen in anaerobic zone (>0.86 mm). Chlorella, Nitrosomonas and Candidatus&#95;Kuenenia were identified as the dominant algae, AOB and AnAOB, with relative abundances of 11.80 %, 19.77 % and 3.07 %, respectively. This layered biofilm benefitted providing a suitable environment for various microorganisms to survive within a complex biofilm., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study effectively coupled anammox and mixotrophic denitrification at a high nitrogen load rate of 6.84 g N/L/d with 40 mg/L Fe(II). Fe(II) enhanced the activity of nitrate reductase, nitrite reductase, and hydrazine dehydrogenase enzymes, facilitating accelerated ATP synthesis. Through electrochemical experiments, interspecies electron transfer processes in coupled system were explored. Fe(II) promoted flavin mononucleotide secretion, enhancing electron-donating and electron-accepting capacity by 2.8 and 1.3 times, respectively. Fe(II) triggered the enrichment of autotrophic denitrifying bacteria (Azospira and Hydrogenophaga), transitioning from single organic nutrient to mixotrophic denitrification. Meanwhile, Fe(II) increased Candidatus&#95;Kuenenia abundance from 35.2 % to 49.0 %, establishing the competitive advantage of anammox bacteria over completed denitrifying bacteria (Comamonas). The synergistic interactions between anammox and various denitrification pathways achieved a nitrogen removal rate of 5.88 g N/L/d, with anammox contribution rate of 88.3 %. This study provides insights into broadening the application of partial denitrification /anammox and electron transfer in multi-bacterial coupling systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study successfully established Iron Sulfide-Mediated mixotrophic Partial Denitrification/Anammox system, achieving nitrogen and phosphorus removal efficiency of 97.26% and 78.12%, respectively, with COD/NO 3 - -N of 1.00. Isotopic experiments and X-ray Photoelectron Spectroscopy analysis confirmed that iron sulfide enhanced autotrophic Partial Denitrification performance. Meanwhile, various sulfur valence states functioned as electron buffers, reinforcing nitrogen and sulfur cycles. Microbial community analysis indicated reduced heterotrophic denitrifiers (OLB8, OLB13) under lower COD/NO 3 - -N, creating more niche space for autotrophic bacteria and other heterotrophic denitrifiers. The prediction of functional genes illustrated that iron Sulfide upregulated genes related to carbon metabolism, denitrification, anammox and sulfur oxidation-reduction, facilitating the establishment of carbon-nitrogen-sulfur cycle. Furthermore, this cycle primarily produced electrons via nicotinamide adenine dinucleotide and sulfur oxidation-reduction processes, subsequently utilized within the electron transfer chain. In summary, the Partial Denitrification/Anammox system under the influence of iron sulfide achieved effient nitrogen removal by expediting electron transfer through the carbon-nitrogen-sulfur cycle., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Sulfate-dependent ammonium oxidation (Sulfammox) is a critical process linking nitrogen and sulfur cycles. However, the metabolic pathway of microbes driven Sulfammox is still in suspense. The study demonstrated that ammonium was not consumed with sulfate as the sole electron acceptor during long-term enrichment, probably due to inhibition from sulfide accumulation, while ammonium was removed at ∼ 10 mg N/L/d with sulfate and nitrate as electron acceptors. Ammonium and sulfate were converted into nitrogen gas, sulfide, and elemental sulfur. Sulfammox was mainly performed by Candidatus Brocadia sapporoensis and Candidatus Brocadia fulgida, both of which encoded ammonium oxidation pathway and dissimilatory sulfate reduction pathway. Not sulfide-driven autotrophic denitrifiers but Candidatus Kuenenia stuttgartiensis converted nitrate to nitrite with sulfide. The results of this study reveal the specialized metabolism of Sulfammox bacteria (Candidatus Brocadia sapporoensis and Candidatus Brocadia fulgida) and provide insight into microbial relationships during the nitrogen and sulfur cycles., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

To reduce the cost of docosahexaenoic acid (DHA) production from Schizochytrium sp., the waste Pichia pastoris was successfully used as an alternative nitrogen source to achieve high-density cultivation during the cell growth phase. However, due to the high oxygen consumption feature when implementing high-density cultivation, the control of both the nitrogen source and dissolved oxygen concentration (DO) at each sufficient level was impossible; thus, two realistic control strategies, including "DO sufficiency-nitrogen limitation" and "DO limitation-nitrogen sufficiency", were proposed. When using the strategy of "DO sufficiency-nitrogen limitation", the lowest maintenance coefficient of glucose (12.3 mg/g/h vs. 17.0 mg/g/h) and the highest activities of related enzymes in DHA biosynthetic routes were simultaneously obtained; thus, a maximum DHA concentration of 12.8 ± 1.2 g/L was achieved, which was 1.58-fold greater than that of the control group. Overall, two-stage feeding control for alternative nitrogen sources is an efficient strategy to industrial DHA fermentation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)