Nitrite Accumulation Research Articles

Effect of humic acid on aerobic denitrification by Achromobacter sp. strain GAD-3

Humic acid (HA), a common natural organic matter, could affect conventional anoxic denitrification. Aim of this study was to investigate effect of HA on the process of aerobic denitrification in Achromobacter sp. GAD-3, an aerobic denitrifying strain. The findings demonstrated that an increase in HA concentrations (≥5mgL-1) promoted the aerobic denitrification process (excluding N2O reduction), manifesting as higher rates of nitrate removal (6.67-11.1mgL-1h-1) and lower levels of nitrite accumulation (30.2-20.7mgL-1). This was attributed to the increased electron transfer activities and denitrifying reductase activities (including NAR, NIR and NOR) facilitated by HA. Accordingly, the expression of denitrification genes such as napA, cnorB, and nirS was enhanced by HA. Nonetheless, the nosZ gene and N2OR activity underwent suppression by HA, which was accountable for N2O emission. It is crucial to understand the HA mechanism towards aerobic denitrifiers for wastewater treatment plants to enhance nitrogen removal.

Possible causes of high nitrite levels in some processed leafy vegetables

Nitrate is a compound that accumulates in the leaves of some leafy vegetables such as spinach, lettuce and rocket, and the European Regulation No. 2023/915 establishes maximum limits, due to some health effects. Nitrite should be absent or present only in traces in leafy vegetables. However, some recent studies demonstrated that not negligible concentrations of nitrite (higher than 50 mg kg−1) may be quantified in leafy vegetables such as spinach and lettuce. In this work, several possible sources of nitrite formation in leafy vegetables, agronomic, environmental, microbiological, and processing have been investigated to evaluate the main factors which led to this accumulation. Microbiological contamination, particularly Enterobacteriaceae, of “fresh-cut” samples of spinach, Swiss chard and wild rocket, has been ascertained as the main source of nitrite accumulation, regardless of other considered factors (agronomic, environmental, processing, etc.). The correlation between Enterobacteriaceae count and nitrite concentration was assessed at 0.98, confirming this one as the main source of nitrite increase, also at very high levels, in processed leafy vegetables.

Unlocking partial nitrification by acidophilic microorganisms in ultra-low strength wastewater: Long-term investigation and genomic insights

Achieving partial nitrification (PN) in low-strength wastewater is still a worldwide issue. In this study, conversion from nitrification to PN was successfully achieved with a self-sustained pH as low as 5.3 and a nitrite accumulation ratio (NAR) of 93.4 % by enriching novel acidic-tolerant functional flora. A strong positive correlation was observed between decreasing pH and NAR (R2 = 0.97). Even with 73.1 % reduction in influent ammonia (from 86.7 to 23.3 mg/L), high dissolved oxygen loading (4.5–5.3 mg/L), and extreme over-aeration, PN efficacy remained robust with the NAR consistently maintained at 90.6 ± 5.3 %. Over 180 days of continuous monitoring, novel ammonia-oxidizing bacteria (AOB) Candidatus Nitrosoglobus became the dominant nitrifying guild (1.65 %) with a specific growth rate of 0.091/day; however, traditional nitrifying bacteria exhibited impaired functionality and bacterial diversity was dramatically reduced. Amo abundance increased by 147 %, while Nxr in nitrifying bacteria dropped below detection limit, preventing complete nitrification. Upregulated electron transport chain genes enhanced electron flow, managing excess oxygen and effectively limiting harmful oxygen radical production, resulting in a desirable nitritation rate of 0.3 kg/m3·d with influent NH4+-N as low as 23.3 mg/L. These regulatory mechanisms boosted NH4+ oxidation efficiency, successfully unlocking scientific barriers associated with PN in ultralow-strength wastewater and laying the groundwork for future PN/A implementation.

Successful operation of nitrifying granules at low pH in a continuous-flow reactor: Nitrification performance, granule stability, and microbial community

Acidic nitrification, as a novel process for treating wastewater without sufficient alkalinity, has received increasing attention over the years. In this study, a continuous-flow reactor with aerobic granular sludge was successful operated at low pH (<6.5) performing high-rate acidic nitrification. Volumetric ammonium oxidation rate of 0.4–1.2 kg/(m3·d) were achieved with the specific biomass activities of 5.8–13.9 mg N/(gVSS·h). Stable partial nitritation with nitrite accumulation efficiency over 85% could be maintained at pH above 6 with the aid of residual ammonium, whereas the nitrite accumulation disappeared when pH was below 6. Interestingly, the granule morphology significantly improved during the acidic operation. The increased secretion of extracellular polymeric substances (especially polysaccharides) suggested a self-protective behavior of microbes in the aerobic granules against acidic stress. 16S rRNA gene sequencing analyses indicated that Candidatus Nitrospira defluvii was always the dominant nitrite-oxidizing bacteria, while the dominant ammonia-oxidizing bacteria shifted from Nitrosomonas europaea to Nitrosomonas mobilis. This study, for the first time, demonstrated the improved stability of aerobic granules under acidic conditions, and also highlighted aerobic granules as a useful solution to achieve high-rate acidic nitrification.

Dynamic pH regulation drives Nitrosomonas for high-rate stable acidic partial nitritation

How to intensify the ammonia oxidation rate (AOR) is still a bottleneck impeding the technology development for the innovative acidic partial nitritation because the eosinophilic ammonia-oxidizing bacteria (AOB), such as Nitrosoglobus or Nitrosospira, were inhibited by the high-level free nitrous acid (FNA) accumulation in acidic environments. In this study, an innovative approach of dynamic acidic pH regulation control strategy was proposed to realize high-rate acidic partial nitritation driven by common AOB genus Nitrosomonas. The acidic partial nitrification process was carried out in a laboratory-scale sequencing batch moving bed biofilm reactor (SBMBBR) for long-term (700 days) to track the effect of dynamic acidic pH on nitrifying bacterial activity. The results indicated that the influent NH4+-N concentration was about 100 mg/L, the nitrite accumulation ratio was exceeding 90%, and the maximum AOR can reach 14.5 ± 2.6 mg N L−1h−1. Although the half-saturation inhibition constant of NOB (KI_FNA(AOB)) reached 0.37 ± 0.10 mg HNO2N/L and showed extreme adaptability in FNA, the inactivation effect of FNA (6.1 mg HNO2N/L) for NOB was much greater than that of AOB, with inactivation rates of 0.61 ± 0.08 h−1 and 0.06 ± 0.01 h−1, respectively. The effluent pH was gradually reduced to 4.5 by ammonia oxidation process and the periodic FNA concentration reached 6.5 mg HNO2N/L to inactivate nitrite-oxidizing bacteria (NOB) without negatively affecting Nitrosomonas during long-term operation. This result provides new insights for the future implementation of high-rate stabilized acidic partial nitritation by Nitrosomonas.

Algal-mediated nitrogen removal and sustainability of algal-derived dissolved organic matter supporting denitrification

Algae-mediated nitrogen removal from low carbon vs. nitrogen (C/N) wastewater techniques has garnered significant attention due to its superior autotrophic assimilation properties. This study investigated the ammonium-N removal potential of four algae species from low C/N synthetic wastewater. Results showed that 95 % and 99 % of ammonium-N are eliminated at initial concentrations of 11.05 ± 0.98 mg/L and 42.51 ± 2.20 mg/L with little nitrate and nitrite accumulation. The compositions of secreted algal-derived dissolved organic matter varied as C/N decreased and showed better bioavailability for nitrate-N removal by Pseudomonas sp. SZF15 without pre-oxidation, achieving an efficiency of 99 %. High-throughput sequencing revealed that the aquatic microbial communities, dominated by Scenedesmus, Kalenjinia, and Micractinium, remain relatively stable across different C/N, aligning with the underlying metabolic pathways. These findings may provide valuable insights into the sustainable elimination of multiple nitrogen contaminants from low C/N wastewater.

Ammonium oxidation from concentrated synthetic wastewater and landfill leachate using partial nitritation in sequencing batch reactor.

Partial nitritation (PN) is a novel treatment for nitrogen removal using aerobic ammonium oxidation with reduced oxygen requirements compared to conventional nitrification. This study evaluated the performance of the PN process and the factors influencing nitrogen removal from landfill leachate. During the reactivation of biomass, the results showed 70% ammonium removal, but only 20% total nitrogen removal. Further analysis showed that low nitrite accumulation and high nitrate production promoted the growth of nitrite-oxidizing bacteria (NOB). The ammonium removal activity after soaking the cultivated biomass in synthetic water and leachate was measured to be 0.57, 0.1, 0.17, and 0.25 g N•g VSS-1•d-1 for synthetic wastewater and leachate soaking for synthetic wastewater, 12 h, 3days, and 7days, respectively. The study found abundant ammonium-oxidizing bacteria (AOB) and NOBs in biomass soaked in synthetic wastewater. However, soaking in leachate promoted AOB growth and inhibited NOB growth making leachate suitable for PN. PRACTITIONER POINTS: The study found that with a longer leachate-soaking period for biomass, ammonium removal activity increases, which in turn increases ammonium conversions during the PN process. Ammonium-oxidizing bacteria (AOB) can acclimate to landfill leachate substrate and grow with a longer soaking period. Nitrite-oxidizing bacteria (NOB) were inhibited by landfill leachate substrate, which is beneficial for nitrite accumulation. Anabolized DO can convert nitrite to nitrate rapidly, which results in higher nitrate accumulation compared to nitrite accumulation. Hence, the DO level has to be sufficiently low to prevent nitrite oxidation and nitrate accumulation.

Response and recovery of nitrifying moving bed biofilm reactor systems exposed to 1°C with varying levels of ammonia starvation

This study investigated the impact of varying total ammonia nitrogen (TAN) feed levels along with water temperature decreases on the performance of nitrifying moving bed biofilm reactor (MBBR) at 1 °C and its recovery at 3 °C. Five MBBR reactors were operated with different TAN concentrations as water temperature decreased from 20 to 3 °C: reactor R1 at 30 mg N/L, reactor R2 at 20 mg N/L, reactor R3 at 15 mg N/L, reactor R4 at 10 mg N/L and reactor R5 at 0 mg N/L. The corresponding biofilm characteristics were also analyzed to understand further nitrifying MBBR under different TAN feeding scenarios. The findings revealed that the higher TAN levels were before reaching 1 °C, the better nitrification performance and the more biomass grew. However, the highest TAN concentration (30 mg N/L) might negatively affect the nitrification performance, the activity of nitrifiers, and the growth of biofilms at 1 °C because of the toxic effects of un-ionized or free ammonia (FA). It was observed that the activities of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were affected by FA concentrations ranging from 0.2 to 0.7 mg N/L at 1 °C, but they could gradually be adapted to such inhibitory environment, with NOB recovering more quickly and robustly than AOB. The study identified 20 mg N/L (67 % of maximum influent TAN at 1 °C in R2 as the optimal TAN feeding concentration, achieving over 90 % TAN removal and a surface area removal rate (SARR) of 0.78 ± 0.02 g N/m2·d at 1 °C. Meanwhile, R2 also exhibited the highest biofilm mass, with total solids at 13.3 mg/carrier and volatile solids at 11.3 mg/carrier. As TAN was removed, nitrite accumulation was observed at 1 °C, and higher influent TAN concentrations prior to 1 °C appeared to delay the accumulation. When water temperature increased from 1 °C to 3 °C, nitrification performance improved significantly in all reactors without nitrite accumulation, and the higher TAN feeding in the previous stage led to faster recovery. Compared with 20 °C, biofilm became thinner and denser at 1 °C and 3 °C. Furthermore, this study revealed significant shifts in microbial community composition and nitrifier abundances in response to changes in water temperature and influent TAN levels. The dominant nitrifiers were identified as Nitrosomonadaceae (AOB) and Nitrospiraceae (NOB). At 1 °C, the nitrifier abundances were significantly correlated with SARRs, FA, and biofilm density. R2, which exhibited the best nitrification performance, maintained higher nitrifier abundances at 1 °C.

Achieving Long-Term Stability of Partial Nitrification and Autotrophic Denitrification in an MABR via Sulfide Dosing.

While partial nitrification (PN) has the potential to reduce energy for aeration, it has proven to be unstable when treating low-strength wastewater. This study introduces an innovative combined strategy incorporating a low rate of oxygen supply, pH control, and sulfide addition to selectively inhibit nitrite-oxidizing bacteria (NOB). This strategy led to a stable PN in a laboratory-scale membrane aerated biofilm reactor (MABR). Over a period of 260 days, the nitrite accumulation ratio exceeded 60% when treating synthetic sewage containing 50 mg NH4+-N/L. Through in situ activity testing and high-throughput sequencing, the combined strategy led to low levels of nitrite-oxidation activity (<5.5 mg N/m2 h), Nitrospira species (relative abundance <1%), and transcription of nitrite-oxidation genes (undetectable). The addition of sulfide led to simultaneous PN and autotrophic denitrification in the single-stage MABR, resulting in over 60% total inorganic nitrogen removal. Sulfur-based autotrophic denitrification consumed nitrite and inhibited NOB conversion of nitrite to nitrate. The combined strategy has potential to be applied in large-scale sewage treatment and deserves further exploration.

Light Enables Partial Nitrification and Algal-Bacterial Consortium in Rotating Biological Contactors: Performance and Microbial Community

Partial nitrification–anaerobic ammonia oxidation represents an innovative nitrogen removal technique, distinguished by its shortened nitrogen removal pathway and reduced energy demands. Currently, partial nitrification is mostly studied in sequential batch reactors, and some of the methods to realize partial nitrification in continuous flow reactors have problems such as complicated operation and management, and can be easily destabilized. This study introduces a novel system utilizing light to establish an algal-bacterial consortium within a partial nitrification framework, where oxygen is supplied by algae and a novel rotating biological contactor (RBC). This approach aims to simplify the control strategy and decrease the energy required for aeration. The results demonstrated that light at an intensity of 200 μmol/(m2·s) effectively inhibited nitrite-oxidizing bacteria (NOB), swiftly stabilizing partial nitrification. In the absence of light, free ammonia (FA) and free nitric acid (FNA) inhibited NOB, with ammonium removal efficiency (ARE) and nitrite accumulation ratio (NAR) at 68.35% and 34.00%, respectively. By day 88, under light exposure, effluent NO2−-N concentrations surged, with ARE and NAR at 64.21% and 69.45%, respectively. By day 98, NAR peaked at 80.28%. The specific oxygen uptake rate (SOUR) of ammonia-oxidizing bacteria (AOB) and NOB outside the disc was 3.24 mg O2/(g MLSS·h) and 0.75 mg O2/(g MLSS·h), respectively. Extracellular polymeric substance (EPS) content initially decreased, then increased, ultimately exceeding pre-light exposure levels. Microbial abundance significantly declined due to light exposure, with Nitrosomonas related-AOB decreasing by 91.88% from 1.6% to 0.13%, and Nitrospira related-NOB decreasing by 99.23% from 5.19% to 0.04%, respectively. The results indicated that both AOB and NOB were inhibited by light, especially NOB. It is a feasible strategy to achieve partial nitrification and algal-bacterial consortia by using light in a rotating biological contactor.

Effects of Inoculating Autochthonous Starter Cultures on Changes of N-Nitrosamines and Their Precursors in Chinese Traditional Fermented Fish during In Vitro Human Digestion.

The objective of this research was to investigate the impact of inoculating autochthonous starter cultures on the alterations in microorganisms, biogenic amines, nitrite, and N-nitrosamines in Chinese traditional fermented fish products (CTFPs) during in vitro human digestion. The results revealed that gastric digestion significantly (p < 0.05) inhibited the proliferation of lactic acid bacteria, yeast, Staphylococcus, and Enterobacteriaceae, whereas various microorganisms proliferated extensively during small intestine digestion. Meanwhile, small intestine digestion could significantly increase (p < 0.05) levels of putrescine, cadaverine, and tyramine. The reduced content observed in inoculated fermentation groups suggests that starter cultures may have the ability to deplete biogenic amines in this digestion stage. Gastric digestion significantly (p < 0.05) inhibited nitrite accumulation in all CTFPs samples. Conversely, the nitrite content increased significantly (p < 0.05) in all groups during subsequent small intestine digestion. However, the rise in the inoculated fermentation groups was smaller than that observed in the spontaneous fermentation group, indicating a potentially positive role of inoculated fermentation in inhibiting nitrite accumulation during this phase. Additionally, gastric digestion significantly (p < 0.05) elevated the levels of N-nitrosodimethylamine (NDMA) and N-nitrosopiperidine in CTFPs. Inoculation with L. plantarum 120, S. cerevisiae 2018, and mixed starter cultures (L. plantarum 120, S. cerevisiae 2018, and S. xylosus 135 [1:1:1]) effectively increased the degree of depletion of NDMA during this digestion process.

Inhibition of zinc ions in sulfur-driven autotrophic denitrification process: What is the behavior of extracellular polymeric substances?

Sulfur-driven autotrophic denitrification (SAD) process is a cost-effective and sustainable method for nitrogen removal from wastewater. However, a higher concentration of zinc ions (Zn(II)) flowing into wastewater treatment plants poses a potential threat to the SAD process. This study examined that a half maximal inhibitory concentration (IC50) of Zn(II) was 7 mg·L−1 in the SAD process. Additionally, the addition of 20 mg·L−1 Zn(II) resulted in a severe accumulation of nitrite to 150.20 ± 6.00 mg·L−1 when the initial concentration of nitrate was 500 mg·L−1. Moreover, the activities of nitrate reductase, nitrite reductase, dehydrogenase and electron transport system were significantly inhibited under Zn(II) stress. The addition of Zn(II) inhibited EPS secretion and worsened electrochemical properties. The result was attributed to the spontaneous binding between EPS and Zn(II), with a ΔG of −17.50 KJ·mol−1 and a binding constant of 1.77 × 104 M−1, respectively. Meanwhile, the protein, fulvic acid, and humic-like substances occurred static quenching after Zn(II) addition, with -OH and -C=O groups providing binding sites. The binding sequence was fulvic acid→protein→humic acid and -OH → -C=O. Zn(II) also reduced the content of α-helix, which was unfavorable for electron transfer. Additionally, the Zn(II) loosened protein structure, resulting in a 50 % decrease in α-helix/(β-sheet+random coil). This study reveals the effect of Zn(II) on the SAD process and enhances our understanding of EPS behavior under metal ions stress.

Full-Scale Demonstration of Nitrogen Removal from Mature Landfill Leachate Using a Two-Stage Partial Nitritation and Anammox Process

The excessive discharge of nitrogen leads to water eutrophication. The partial nitritation and anammox (PN/A) process is a promising technology for biological nitrogen removal in wastewater treatment. However, applying it to mature landfill leachate (MLL) faces challenges, as the toxic substances (e.g., heavy metal) within MLL inhibit the activity of anammox bacteria. Therefore, most previous studies focused on diluted, pretreated, or chemically adjusted MLL. This study demonstrated at full scale that the two-stage PN/A process can treat raw MLL. Initially, the operational issue of sludge floatation resulted in rapid biomass loss with overflow discharging, which selectively suppresses nitrite-oxidizing bacteria (NOB), promoting the achievement of nitrite accumulation. After that, the NOB suppression was self-sustained by the high in situ free ammonia concentration, i.e., 26.2 ± 15.9 mg N/L. In the subsequent anammox tank, nitrogen removal primarily occurred via the anammox process, complemented by denitrification, achieving total nitrogen removal efficiency exceeding 72%. In addition, the nitrogen removal capacity of this system was significantly influenced by temperature with the nitrogen-loading rate above 0.4 kg N/m3/d at 38 °C and approximately 0.1 kg N/m3/d at 21 °C. The optimization of system operation, such as gradually increasing MLL content, remains necessary to enhance nitrogen removal capacity further.

Enhanced anammox performance under lower nitrite accumulation in modified partial nitritation-anammox (PN/A) process

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 NH4+-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.

Coupling Nitrate-Dependent Anaerobic Ethane Degradation with Anaerobic Ammonium Oxidation.

The microbial oxidation of short-chain gaseous alkanes (SCGAs, consisting of ethane, propane, and butane) serves as an efficient sink to mitigate these gases' emission to the atmosphere, thus reducing their negative impacts on air quality and climate. "Candidatus Alkanivorans nitratireducens" are recently found to mediate nitrate-dependent anaerobic ethane oxidation (n-DAEO). In natural ecosystems, anaerobic ammonium-oxidizing (anammox) bacteria may consume nitrite generated from nitrate reduction by "Ca. A. nitratireducens", thereby alleviating the inhibition caused by nitrite accumulation on the metabolism of "Ca. A. nitratireducens". Here, we demonstrate the coupling of n-DAEO with anammox in a laboratory-scale model system to prevent nitrite accumulation. Our results suggest that a high concentration of ethane (6.9-7.9%) has acute inhibition on anammox activities, thus making the coupling process a significant challenge. By maintaining ethane concentrations within the range of 1.7-5.5%, stable ethane and ammonium oxidation, nitrate reduction, and dinitrogen gas generation without nitrite accumulation were finally achieved. After the accomplished coupling of n-DAEO with anammox, nitrate reduction rates increased by 8.1 times compared to the rate observed with n-DAEO alone. Microbial community profiling via 16S rRNA gene amplicon sequencing showed "Ca. A. nitratireducens" (6.6-12.9%) and anammox bacteria "Candidatus Kuenenia" (3.4-5.6%) were both dominant in the system, indicating they potentially form a syntrophic partnership to jointly contribute to nitrogen removal. Our findings offer insights into the cross-feeding interaction between "Ca. A. nitratireducens" and anammox bacteria in anoxic environments.

Nitrogen removal characteristics and Cr(VI) tolerance mechanisms of heterotrophic nitrifying bacterium Pseudomonas putida strain LX1

To solve the severe inhibition effect of nitrification process by heavy metals, necessitating the discovery of heavy metals-resistant nitrifying bacteria. In this study, a highly efficient bacterium, Pseudomonas putida strain LX1, capable of heterotrophic nitrification–aerobic denitrification (HN-AD) and resistant to hexavalent chromium (Cr(VI)) was isolated from activated sludge. Strain LX1 exhibited excellent growth activity and achieved efficient NH4+-N removal rates (94.07%–83.36 %) under 0–25 mg/L Cr(VI) stress. The heterotrophic nitrification ability of strain LX1 was superior to that of aerobic denitrification, and there was no accumulation of nitrite or nitrate during HN-AD process. Moreover, Three-dimensional excitation-emission matrix and Fourier-transform infrared spectroscopy analysis indicated that strain LX1 secreted more extracellular polymeric substances with negatively charged functional groups to bind Cr(VI) and alleviate its toxicity. Enzymatic analysis showed that strain LX1 countered the oxidative damage of reactive oxygen species by promoting the activities of oxidative dismutase and catalase. Furthermore, transcriptomic analyses revealed that the upregulation of genes associated with heavy metal substrate transporter proteins and antioxidant systems in strain LX1 contributed to the resistance against the high biotoxicity caused by Cr(VI). This study is important for understanding the heavy metal response mechanisms of HN-AD strains and its potential application in practical wastewater treatment.

Effects of Weak Electric Fields on the Denitrification Performance of Pseudomonas stutzeri: Insights into Enzymes and Metabolic Pathways.

Enhanced denitrification has been reported under weak electric fields. However, it is difficult to investigate the mechanism of enhanced denitrification due to the complex interspecific interactions of mixed-culture systems. In this study, Pseudomonas stutzeri, capable of denitrification under anaerobic conditions, was selected for treating low COD/N (2.0, ratio between concentration of chemical oxygen demand and NO3--N) artificial wastewater under constant external voltages of 0.2, 0.4, and 0.6 V. The results revealed that P. stutzeri exhibited the highest efficiency in nitrate reduction at 0.2 V. Moreover, the maximum nitrate removal rate was 15.96 mg/(L·h) among the closed-circuit groups, 19.39% higher than that under the open-circuit group. Additionally, a notable reduction in nitrite accumulation was observed under weak electric fields. Enzyme activity analysis showed that the nitrate reductase activities were significantly increased among the closed-circuit groups, while nitrite reductase activities were inhibited. Transcriptomic analysis indicated that amino acid metabolism, carbohydrate metabolism, and energy metabolism were increased, enhancing the resistance of P. stutzeri to environmental stress and the efficiency of carbon source utilization for denitrification. The current study examined the impacts of weak electric fields on enzyme activities and microbial metabolic pathways and offers valuable insights into the mechanism by which denitrification is enhanced by weak electric fields.

Intermittent hydroxylamine dosing to strengthen stability of partial nitrification and nitrogen removal efficiency through continuous-flow anaerobic–aerobic-anoxic reactor treating municipal wastewater

Intermittent hydroxylamine (NH2OH) dosing strategy was applied to enhance the stability of partial nitrification and total nitrogen (N) removal efficiency (TNRE) in a continuous-flow process. The results showed 2 mg/L of NH2OH dosing (once every 6 h) could maintain stably partial nitrification with nitrite accumulation rate (NAR) of 91.6 % and TNRE of 92.6 %. The typical cycle suggested NH2OH dosing could promote simultaneous nitrification–denitrification (SND) and endogenous denitrification (END) while inhibit exogenous denitrification (EXD). Nitrification characteristics indicated the NH2OH dosing enhanced stability of partial nitrification by suppressing specific nitrite oxidation rate (SNOR), Nitrospira and nitrite oxidoreductase enzyme (Nxr). The microbial community suggested the aerobic denitrfiers, denitrifying glycogen accumulating organisms (DGAOs) and traditional denitrfiers were the potential contributor for advanced N removal. Moreover, NH2OH dosage was positively associated with NAR, SND and END. Overall, this study offers a feasible strategy to maintain sustainably partial nitrification that has great application potential.

Effect of Carbon Source on Endogenous Partial Denitrification Process: Characteristics of Intracellular Carbon Transformation and Nitrite Accumulation

This study focused on the start-up and operating characteristics of the endogenous partial denitrification (EPD) process with different carbon sources. Two sequencing batch reactors (SBRs) with sodium acetate (SBR1#) and glucose (SBR2#) as carbon sources were operated under anaerobic/oxic (A/O) and anaerobic/anoxic/oxic (A/A/O) modes successively for 240 d. The results showed that COD removal efficiency reached 85% and effluent COD concentrations were below 35 mg/L in both SBRs. The difference was that faster absorption and transformation of sodium acetate was achieved compared to glucose (COD removal rate (CRR) was 7.54 &gt; 2.22 mgCOD/(L·min) in SBR1# compared to SBR2#). EPD could be started up with sodium acetate and glucose as carbon sources, respectively, and desirable high nitrite accumulations were both obtained at influent NO3−−N (NO3−-Ninf) increased from 20 to 40 mg/L with nitrate-to-nitrite transformation ratio (NTR) and specific NO3−-N deduction rate (rNa) of 88.4~90% and 2.41~2.38 mgN/(gVSS·h), respectively. However, at NO3−-N of 50~60 mg/L, both the NTR and rNa in SBR1# were higher compared to SBR2# (86.5% &gt; 83.9% and 1.58 &gt; 1.20 mgN/(gVSS·h), respectively). Hereafter, when NO3−-N was increased by 70~90 mg/L, lower NTR and rNa were observed in SBR1# than in SBR2# (72% and 78%, 1.16 and 1.32 mgN/(gVSS·h), respectively). Additionally, similar internal carbon transformations were observed to drive EPD for NO2−−N accumulation, especially for higher and faster carbon transformation with sodium acetate as carbon source compared to glucose. However, precise control of anoxic time as the peak point of nitrite (TNi,max) was still the key to achieve high NO2−−N accumulation.

How mineral induced antibiotic transformation products impact bacterial growth and denitrification activity.

The abiotic transformations of quinolones and tetracyclines facilitated by redox-active minerals has been studied extensively, however limited information is available regarding the antimicrobial activity and toxicity of their resultant transformation products. In this study, we first investigated the mechanisms underlying the transformation of two commonly used antibiotics, ciprofloxacin (CIP) and tetracycline (TC), by the ubiquitous redox soil mineral, birnessite (MnO2). Subsequently, we evaluated the impact of these transformation products on both the growth and activity of the environmental denitrifier Pseudomonas veronii. Following the reaction with birnessite, four transformation products for CIP and five for TC were identified. Remarkably, the antibacterial activity of both CIP and TC was lost upon the formation of transformation products during their interaction with birnessite. This loss of antimicrobial efficacy was associated with specific chemical transformations, such as the opening of the piperazine ring for CIP and hydroxylation and demethylation for TC. Interestingly, denitrifying activity, quantified in terms of nitrate reduction rates, remained unaffected by both CIP and TC at low concentrations that did not impact bacterial growth. However, under certain conditions, specifically at low concentrations of CIP, the second step of denitrification-nitrite reduction-was hindered, leading to the accumulation of nitrite. Our findings highlight that the transformation products induced by the mineral-mediated reactions of CIP or TC lose the initial antibacterial activity observed in the parent compounds. This research contributes valuable insights into the intricate interplay between antibiotics, redox-active minerals, and microbial activity in environmental systems.