Comprehensive study on pilot nitrification-sludge fermentation coupled denitrification system with extended sludge retention time.

The sludge fermentation-coupled denitrification process, utilized for sludge reduction and nitrogen removal from wastewater, is frequently hindered by its hydrolysis step's efficacy. This study addresses this limitation by extending the sludge retention time (SRT) to 120 days. As a result, the nitrate removal efficiency (NRE) of the nitrification-sludge fermentation coupled denitrification (NSFD) pilot system increased from 67.1 ±â€¯0.2 % to 96.7 ±â€¯0.1 %, and the sludge reduction efficiency (SRE) rose from 40.2 ±â€¯0.5 % to 62.2 ±â€¯0.9 %. Longer SRT enhanced predation and energy dissipation, reducing intact cells from 99.2 % to 78.0 % and decreasing particle size from 135.2 ±â€¯4.6 µm and 19.4 ±â€¯2.1 µm to 64.5 ±â€¯3.5 µm and 15.5 ±â€¯1.6 µm, respectively. It also created different niches by altering the biofilm's adsorption capacity, with interactions between these niches driving improved performance. In conclusion, extending SRT optimized the microbial structure and enhanced the performance of the NSFD system.

Interspecies cooperation-driven photogenerated electron transfer processes and efficient multi-pathway nitrogen removal in the g-C3N4-anammox consortia biohybrid system.

Photocatalytic materials-microbial biohybrid systems pave the way for solar-driven wastewater nitrogen removal. In this study, interspecies cooperation in photogenerated electron transfer and efficient nitrogen removal mechanism in the g-C3N4-anammox consortia biohybrid system were first deciphered. The results indicated that the essential extracellular electron carriers (cytochrome c and flavin) for anammox genomes were provided by associated bacteria (BACT3 and CHLO2). This cooperation, regulated by the ArcAB system and electron transfer flavoprotein, made anammox bacteria the primary photogenerated electron sink. Furthermore, an efficient photogenerated electron harness was used to construct a reductive glycine pathway (rGlyP) in anammox bacteria inventively, which coexisted with the Wood-Ljungdahl pathway (WLP), constituting a dual-pathway carbon fixation model, rGlyP-WLP. Carbon fixation products efficiently contributed to the tricarboxylic acid cycle, while inhibiting electron diversion in anabolism. Photogenerated electrons were targeted channeled into nitrogen metabolism-available electron carriers, enhancing anammox and dissimilatory nitrate reduction to ammonium (DNRA) processes. Moreover, ammonia assimilation by the glycine cleavage system in rGlyP established an alternative ammonia removal route. Ultimately, multi-pathway nitrogen removal involving anammox, DNRA, and rGlyP achieved 100 % ammonia removal and 94.25 % total nitrogen removal efficiency. This study has expanded understanding of anammox metabolic diversity, enhancing its potential application in carbon-neutral wastewater treatment.

Anammox Coupled with Photocatalyst for Enhanced Nitrogen Removal and the Activated Aerobic Respiration of Anammox Bacteria Based on cbb3-Type Cytochrome c Oxidase.

This study introduced photogenerated electrons into the anammox system by coupling them to a g-C3N4 nanoparticle photocatalyst. A high nitrogen removal efficiency (94.25%) was achieved, exceeding the biochemical limit of 89% imposed by anammox stoichiometry. Photogenerated electrons boosted anammox metabolic activity by empowering key enzymes (NIR, HZS, and WLP-related proteins) and triggered rapid algal enrichment by enhancing the algal Calvin cycle, thus developing multiple anammox-algae synergistic nitrogen removal processes. Remarkably, the homologous expression of cbb3-type cytochrome c oxidase (CcO) in anammox bacteria was discovered and reported in this study for the first time. This conferred aerobic respiration capability to anammox bacteria and rendered them the principal oxygen consumer under 7.9-19.8 mg/L dissolved oxygen, originating from algal photosynthesis. Additionally, photogenerated electrons selectively targeted the cb1 complex and cbb3-type CcO as activation sites while mobilizing the RegA/B regulatory system to activate the expression of cbb3-type CcO. Furthermore, cbb3-type CcO blocked oxidative stress in anammox by depleting intracellular oxygen, a substrate for reactive oxygen species synthesis. This optimized the environmental sensitivity of anammox bacteria and maintained their high metabolic activity. This study expands our understanding of the physiological aptitudes of anammox bacteria and provides valuable insights into applying solar energy for enhanced wastewater treatment.

Metagenomic insights into responses of microbial population and key functional genes to fulvic acid during partial nitritation.

The long-term impact of fulvic acid (FA) on partial nitritation (PN) system was initially examined in this study. The obtained results revealed that the FA lower than 50 mg/L had negligible effect on the nitrite accumulation rate (NAR nearly 100%) and ammonium removal rate (ARR 56.85%), while FA over 50 mg/L decreased ARR from 56.85% to 0.7%. Sludge characteristics analysis found that appropriate FA (<50 mg/L) exposure promoted the settling performance and granulation of PN sludge by removing Bacteroidetes and accumulating Chloroflexi. The analysis of metagenomics suggested that the presence of limited FA (0-50 mg/L) stimulated the generation of NADH, which favors the denitrification and nitrite reduction. The negative impact of FA on the PN system could be divided into two stages. Initially, limited FA (50-120 mg/L) was decomposed by Anaerolineae to stimulate the growth and propagation of heterotrophic bacteria (Thauera). Increasing heterotrophs competed with AOB (Nitrosomonas) for dissolved oxygen, causing AOB to be eliminated and ARR to declined. Subsequently, when FA dosage was over 120 mg/L, Anaerolineae were inhibited and heterotrophic bacteria reduced, resulting in the abundance of AOB recovered. Nevertheless, the ammonium transformation pathway was suppressed because genes amoABC and hao were obviously reduced, leading to the deterioration of reactor performance. Overall, these results provide theoretical guidance for the practical application of PN for the treatment of FA-containing sewage.

Multi-Omics Analysis Reveals the Nitrogen Removal Mechanism Induced by Electron Flow during the Start-up of the Anammox-Centered Process.

Significant progress in understanding the key enzymes or species of anammox has been made; however, the nitrogen removal mechanism in complex coupling systems centered on anammox remains limited. In this study, by the combination of metagenomics-metatranscriptomics analyses, the nitrogen removal in the anammox-centered coupling system that entails partial denitrification (PD) and hydrolytic acidification (HA, A-PDHA) was elucidated to be the nitrogen transformation driven by the electron generation-transport-consumption process. The results showed that a total nitrogen (TN) removal efficiency of >98%, with a TN effluence of <1 mg/L and a TN removal contribution via anammox of >98%, was achieved after 59 days under famine operation and alkaline conditions during the start-up process. Further investigation confirmed that famine operation promoted the activity of genes responsible for electron generation in anammox, and increased the abundance or expression of genes related to electron consumption. Alkaline conditions enhanced the electron generation for PD by upregulating the activity of glyceraldehyde 3-phosphate dehydrogenase and strengthened electron transfer by increasing the gene encoding quinone pool. Altogether, these variations in the electron flow led to efficient nitrogen removal. These results improve our understanding of the nitrogen removal mechanism and application of the anammox-centered coupling systems in treating nitrogen wastewater.

Microbial driving mechanism for simultaneous removal of nitrogen and phosphorus in a pure anammox reactor under ferrous ion exposure.

The mechanisms of Fe2+ on nitrogen and phosphorus removal and functional bacterial competition in anammox systems was investigated. Under 0.12 mM Fe2+, the performance of nitrogen and phosphorus removal increased by 10.08 % and 151.91 %, respectively, compared with the control stage. Phosphorus removal was achieved through extracellular polymeric substance (EPS) induced biomineralization to form Fe-P minerals, and functional group COC in EPS played a critical role. T-EPSs was the major nucleation site due to it maintaining the supersaturated state (saturation index > 0) of Fe-P minerals for a long time. Population succession showed that Fe2+ weakened the competition between heterotrophic denitrifier (Denitrasoma) and anammox microbe (Candidatus Brocadia) for space and substrates, which was favorable for the enrichment of anammox biomass. Moreover, the variation in gene abundance (such as Hao, Cyt c, and Nir) indicated that Fe2+ improved electron behaviors (generation, transport, and consumption) during the nitrogen metabolism of anammox systems.

Metagenomics illuminated the mechanism of enhanced nitrogen removal and vivianite recovery induced by zero-valent iron in partial-denitrification/anammox process.

In this study, a novel strategy of zero-valent iron (ZVI) combined with acetic acid was proposed to optimize partial-denitrification/anammox (PD/A) process, and enhanced nitrogen removal mechanism was elucidated through metagenomics. Results showed that the optimal nitrogen and phosphorus removal were as high as 99.50% and 98.37%, respectively, with vivianite being precipitated as the main byproduct. The occurrence of Feammox was a crucial link for enhanced ammonia removal and vivianite recovery. Metagenomic analysis further certified that long-term acclimation of optimization strategy triggered DNRA-based nitrate reducing genes (narY/Z and nrfA) assigned to Candidatus Brocadia, which allow direct uptake of nitrate by the anammox. Additionally, ZVI might act as a new electron donor to decrease organics dependence of PD by reducing the abundance of genes for electron production involved in carbon metabolism. However, FA addition enhanced the relative abundances of genes involved in anammox including nitrogen reduction and oxidation, thereby accelerating nitrogen removal.

Impact mechanism and performance enhancement of ultrasound on ZVI-anammox system.

The zero-valent iron-anaerobic ammonium oxidation (ZVI-anammox) system has received widespread attention due to its excellent nitrogen removal performance and user-friendly operation. However, its disadvantages include a short service life, high ZVI consumption, and poor system stability. The use of ultrasound as a physical method is increasing in various water treatment processes. In this study, a series of batch tests were conducted to obtain the best ultrasonic parameter and explore the comprehensive effects of ultrasound on a ZVI-anammox system. The highest specific anammox activity of the ZVI-anammox system was found to be 2.88 mg total nitrogen/g of volatile suspended solids/h after ultrasonic treatment (0.2 w/mL, 5 min), which was 37.85% higher than a control group. Additionally, the service life of ZVI extended by 28.57% and the total nitrogen removal efficiency changed from 58.03-72.08 to 63.92-78.33% under ultrasonic irradiation. These phenomena were related to the mechanical force and cavitation of ultrasonic waves. Judging from the characteristics of sludge and ZVI, ultrasound can promote anammox sludge granulation, ease ZVI passivation, and enhance the stability of the entire system. This paper also briefly discusses the impact mechanisms of ultrasound on the ZVI-anammox system. In brief, ultrasound destroys the surface structure of ZVI and thus provides numerous attachment points for microorganisms that improve the performance of the entire system. The proposed ultrasound combined with ZVI is a novel method that has potential for use in large-scale engineering applications in actual sewage treatment after comprehensive analysis.

Anammox-synchronous zero-valent iron oxidation promoting synergistic nitrogen and phosphorus removal from wastewater.

In this study, a novel process, anammox-synchronous zero-valent iron (ZVI) oxidationin whichnitrate byproductsare usedfor in situoxidization of ZVIwas first developed to simultaneously remove nitrogen and phosphorus from wastewater. The optimal ZVI dosage of 4 g/L significantly improved the nitrogen removal efficiency to 86.02 ± 1.98%, with the highest phosphorus removal efficiency enhanced from 39.62% to 98.97%. Several analytical techniques revealed that iron phosphate minerals formed by biologically induced mineralization promoted the phosphorus removal of the system and enhanced the settleability of granules. Candidatus Brocadia was the main anammox functional bacteria, which could promote the formation of iron phosphorus minerals. The increase of key functional genes involved in denitrification, especially narG, played a pivotal role in reducing nitrate to improve nitrogen removal performance. In addition, abundance regulation of gene fur enabled anammox bacteria always maintain high activity under different pH and ZVI dosages.

Rapid start-up strategy of partial denitrification and microbially driven mechanism of nitrite accumulation mediated by dissolved organic matter.

The rapid start-up of Partial denitrification (PD; nitrate to nitrite) was investigated based on the analysis of microbially driven mechanism of nitrite accumulation mediated by Dissolved organic matter (DOM) in this study. The nitrate to Nitrite transformation ratio (NTR) > 90% and effluent nitrate < 5 mg/L were achieved in 17 days by feeding with lower nitrate of ~ 35 mg/L and removing the idling period. And the enhanced nitrite accumulation when applying the above strategy is related to the decreased utilization of the aliphatic DOM during nitrite reduction process. Additionally, the rapid enriched Thauera and OLB13 (37.21%) and inhibited norank_f__Blastocatellaceae (2.86%), and the increased disparity (2.0-fold) between the genes involved in nitrite generation (e.g., narH) and for nitrite reduction (e.g., nirK) jointly contributed to PD start-up. While the genes (e.g., DLD) related to producing electrons from aliphatic DOM also up-regulated by 0.1-fold, which led to the increased nitrate removal and NTR.

Effect of fulvic acid on bioreactor performance and on microbial populations within the anammox process.

The long-term effect of fulvic acid (FA) on bioreactor performance and on microbial populations within the anammox process were firstly investigated in this study. The average nitrogen removal rate showed an upward trend when the influent TOC concentration of FA was 25.2-65.1 mg/L. However, when FA was increased to 80.3 mg/L, the reactor performance was slightly inhibited. In addition, judging from the particle size and settling properties, FA can promote anammox sludge granulation. After 53 days of exposure to FA, the genus Anaerolineaincreased in number, while Denitratisoma decreased. Candidatus Jettenia and Candidatus Kuenenia survived and enriched in the changed environment, potentially due to the interaction between anammox bacteria and some heterotrophic bacteria, which could protect anammox bacteria from adverse environments. These results indicate that FA can change the bacterial community and trigger different microbial interaction mechanisms within the anammox reactor.