Partial Denitrification Process Research Articles

Efficient partial denitrification-anammox process enabled by a novel denitrifier with truncated nitrite reduction pathway

Partial denitrification coupled with anaerobic ammonium oxidation (PD-anammox) is a promising technology for cost-effective nitrogen removal from wastewater. Nitrite availability is crucial to anammox performance but often limited by the slow partial denitrification process. Here we report an efficient PD-anammox system driven by the novel denitrifier Bacillus velezensis C1-3 with truncated nitrite reduction pathway. Whole-genome sequencing analysis revealed that the lack of nitrite reductase genes nirS/nirK and norBC in strain C1-3 enabled nitrite accumulation without the need for precise control of carbon dosage. By coupling it with anammox sludge, over 79% total nitrogen (TN) removal was stably achieved, under a TN loading rate of 660mg/L/d and a carbon/nitrogen ratio below 1.0. Mechanism explorations indicate that the niche differentiation of C1-3 and anammox bacteria facilitated their mutualism while avoiding nitrite competition. This study demonstrates a novel strategy for establishing efficient PD-anammox process by harnessing the unique metabolic deficiency of denitrifiers, shedding light on the development of stable and sustainable biological nitrogen removal technologies with minimal carbon footprint.

Sulfamethoxazole exposure shifts partial denitrification to complete denitrification: Reactor performance and microbial community

This study elucidated the influence on a partial denitrification (PD) system under 0–1 mg/L sulfamethoxazole (SMX) stress in a sequencing batch reactor. The results showed that the nitrite accumulation rate (NAR) significantly (P ≤ 0.01) decreased from 68.68 ± 9.00% to 49.05 ± 11.68%, while the total nitrogen removal efficiency significantly (P ≤ 0.001) increased from 23.19 ± 4.42% to 31.36 ± 2.73% in presence of SMX. The results indicated that SMX exposure switched the PD process to complete denitrification through the deterioration of the nitrite accumulation and the promotion of further nitrite reduction. The SMX removal loading rate increased from 0.21 ± 0.04 to 5.03 ± 0.77 mg-SMX/(g-MLVSS·d) with the extended reactor operation under SMX stress. Low SMX concentration exposure increased extracellular polymeric substances (EPS) content from 107.69 ± 20.78 mg/g-MLVSS (0.05 mg-SMX/L) to 123.64 ± 9.66 mg/g-MLVSS (0.5 mg-SMX/L), while EPS secretion was inhibited under high SMX concentration exposure (i.e., 1 mg-SMX/L). Moreover, SMX exposure promoted the synthesis of aromatic protein-like compounds and changed the functional groups as revealed by EEM and FTIR analysis. Additionally, SMX exposure significantly shifted the microbial community structures at both phylum and genus levels. Particularly, the abundance of Thauera, i.e., functional bacteria related to PD, considerably decreased from 41.69% to 11.62% after SMX exposure, whereas the abundances of Denitratisoma and SM1A02 significantly rose under different SMX concentrations. These outcomes hinted that the addition of SMX resulted in the shifting of partial denitrification to complete denitrification.

Molecular evidence of internal carbon-driven partial denitrification in a mainstream pilot A-B system coupled with side-stream EBPR treating municipal wastewater

Achieving mainstream short-cut nitrogen removal via nitrite has become a carbon and energy efficient way, but still remains challenging for low-strength municipal wastewaters. This study integrated sidestream enhanced biological phosphorus removal system in a pilot-scale adsorption/bio-oxidation (A-B) process (named A-B-S2EBPR system) and nitrite accumulation was successfully achieved for treating the municipal wastewater. Nitrite could accumulate to 5.5 ± 0.3 mg N/L in the intermittently aerated tanks of B-stage with the nitrite accumulation ratio (NAR) of 79.1 ± 6.5 %. The final effluent concentration and removal efficiency of total inorganic nitrogen (TIN) were 4.6 ± 1.8 mg N/L and 84.9 ± 5.6 %, respectively. In-situ process performance of nitrogen conversions, routine batch nitrification/denitrification activity tests and functional gene abundance of nitrifiers collectively suggested that the nitrite accumulation was mainly caused by partial denitrification rather than out-selection of nitrite oxidizing bacteria (NOB). Moreover, the single-cell Raman spectroscopy analysis first demonstrated that there was a specific microbial population that could utilize polyhydroxyalkanoates (PHA) as the potential internal carbon source during the partial denitrification process. The integration of S2EBPR brings unique features to the conventional A-B process, such as extended anaerobic retention time, lower oxidation–reduction potential (ORP), much higher and complex volatile fatty acids (VFAs) etc., which can largely reshape the microbial communities. The dominant genera were Acinetobacter and Comamonadaceae, which accounted for (17.8 ± 15.5)% and (6.7 ± 3.4)%, respectively, while the relative abundance of conventional nitrifiers was less than 0.2%. This study provides insights into phylogenetic and phenotypic shifts of microbial communities when incorporating S2EBPR into the sustainable A-B process to achieve mainstream short-cut nitrogen removal.

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 > 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% > 83.9% and 1.58 > 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.

Rapid enrichment of AnAOB with a novel vermiculite/tourmaline modification technology for enhanced DEAMOX process

The DEnitrifying AMmonium OXidation (DEAMOX) has been proven to be a promising process treating contaminated surface water containing ammonia and nitrate, while the enrichment of the slow-growing anammox bacteria (AnAOB) remains a challenge. In this study, a novel polyurethane-adhesion vermiculite/tourmaline (VTP) modified carrier was developed to achieve effective enrichment of AnAOB. The results demonstrated that the VTP-1 (vermiculite: tourmaline = 1:1) system exhibited the greatest performance with the total nitrogen removal efficiency reaching 87.6% and anammox contributing 63% to nitrogen removal. Scanning electron microscope analysis revealed the superior biofilm structure of the VTP-1 carrier, providing attachment for AnAOB. The addition of VTP-1 promoted the secretion of EPS (extracellular polymeric substances) by microorganisms, which increased to 85.34 mg/g VSS, contributing to the aggregation of anammox cells. The favorable substrate microenvironment created by NH4+ adsorption and NO2− supply via partial denitrification process facilitated the growth of AnAOB. The relative abundance of Candidatus Brocadia and Thauera increased from 0.04% to 0.3%–1.03% and 2.06% in the VTP-1 system, respectively. This study sheds new light on the anammox biofilm formation and provides a valid approach to initiate the DEAMOX process for low nitrogen polluted water treatment.

Mechanisms of partial denitrification in an integrated fixed-film activated sludge (IFAS) system: From progressive startup to stabilization

The rapid startup and long-term stability of partial denitrification (PD) processes are essential prerequisites for successful PD/Anammox systems. Bioaggregates play a significant role in impacting mass transfer efficiency and shaping the microbial community structure within the PD process. Thus, the startup and stable operation of the PD process were investigated in an integrated fixed-film activated sludge (IFAS) by inoculating activated sludge and biofilm from anoxic tanks of a wastewater treatment plant. During the initial stage of startup, a sudden increase in NO3−-N concentration induced a rapid enzyme stress response, leading to a significant increase in total NaR activity (1.38 ± 0.0 μmol·(L·min)−1 to 25.94 ± 0.23 μmol·(L·min)−1), concomitant with the inhibition of total NiR activity (13.93 ± 0.48 μmol·(L·min)−1 to 4.25 ± 0.26 μmol·(L·min)−1), thereby inducing the rapid accumulation of NO2−-N (13.39 mg·L−1) within 6 days. During the later stage of startup, enzyme stress response was subsided but the partial denitrifiers were gradually enriched, consequently leading to continuous accumulation of NO2−-N. The results of ex-situ activity showed that flocs exhibit superior PD performance in contrast to biofilm throughout the entire operational period. Meanwhile, the total extracellular polymeric substances (EPS) content and community structure in both flocs and biofilm indicated that the migration of EPS-secreted partial denitrifiers (Thauera) from flocs to biofilm was obliged by substrate deficiency during the stable operating stage (days 35–100). Therefore, the startup of PD may be sequentially mediated by two distinct mechanisms, enzyme stress response and functional bacterial enrichment, with microbial community migration occurring during the stable phase.

Interactions and coupling mechanisms between Anammox, partial denitrification (PD) and hydroxyapatite (HAP) biomineralization in a PD/A-HAP system

In this study, hydroxyapatite (HAP) crystallization was introduced into a partial denitrification/Anammox (PD/A) process to develop PD/A-HAP granules and achieve simultaneous removal of nitrogen and phosphorus from wastewater. During the formation of PD/A-HAP granules, an unbalanced relationship between the biomass growth and phosphorus crystallization was observed, which resulted in a mass transfer inhibition inside the system. PD and Anammox activities were both impacted by the impeded nutrient contact, and PD process was revealed to be more susceptible to inhibition. Through the regulation of COD and NO3–-N, a dynamic balance between microbial proliferation and phosphorus crystallization was achieved. As a result, high nitrogen removal efficiency (NRE) was attained at 97.8±1.2 % in the final stage of operation, accompanied by a phosphorus removal efficiency (PRE) of 59.4±2.9 %. Furthermore, the recovered phosphorus was transformed into HAP crystals and encapsulated by the PD/A biomass to form granules, which greatly promoted the settling performance of the sludge. This study underlines the importance of reconciling biomass and HAP growth for a strengthened PD/A granular sludge and an enhanced nitrogen removal system. The constructed PD/A-HAP process will expand the application of PD/A with its ability to remove phosphorus while efficiently removing nitrogen.

Study of bacterial population dynamics in seed culture developed for ammonia reduction from synthetic wastewater.

The waterbodies have been polluted by various natural and anthropogenic activities. The aquatic waste includes ammonia as one of the most toxic pollutants. Several biological treatment systems involving anoxic and semi anoxic bacteria have been proposed for reducing nitrogen loads from wastewater and increasing the efficiency and cost effectiveness. These bacteria play a vital role in the processes involved in the nitrogen cycle in nature. However, the enrichment, sustainability and identification of bacterial communities for wastewater treatment is an important aspect. Most of the chemolithotrophs are unculturable hence their identification and measurement of abundance remains a challenging task. In this study the different bacteria involved in total nitrogen removal from the wastewater are enriched for 700days under anoxic condition. The synthetic wastewater containing 0.382g/L of ammonium chloride was used. Molecular identification of the bacteria involved in various steps of the nitrogen cycle was carried out based on amplification of functional genes and 16S rRNA gene Polymerase chain reaction followed by DNA sequencing. Change in the abundance of chemolithotrophs was studied using qPCR. The mutual growth of various nitrifiers along with anaerobic bacteria were identified by molecular characterisation of DNA at various time intervals with the different genes involved in the nitrogen cycle. Nitrosomonas species like Nitrosomonas europaea were identified throughout the batch scale studies possessing the genes associated with ammonia oxidizing bacteria and nitrite oxidizing bacteria which act as a complete ammonia oxidizer. The uncultured species of Nitrospira and anammox bacteria were also observed which predicts the coexistence of the anammox and comammox bacteria in a batch scale study. The coexistence of the semi anoxic and anoxic bacteria helped in the growth of these bacteria for a longer duration of time. The nitrite produced by the comammox during nitrification can be utilized by anammox as an electron carrier. The other species of denitrifiers like Pseudomonas denitrificans and Aminobacter aminovorans were also observed. It is concluded that the enrichment of semi anoxic and anoxic bacteria was faster with the increase in growth of the bacteria involved in nitrification, comammox, anammox and partial denitrification process. The bacterial growth is enhanced and the efficiency is increased which can be further used in the development of small pilot scale bioreactor for total nitrogen removal.

Efficient nitrogen removal from real municipal wastewater and mature landfill leachate using partial nitrification-simultaneous anammox and partial denitrification process

Anaerobic ammonia oxidation (anammox) of municipal wastewater is a research focus, especially the combined treatment with mature landfill leachate is a current research hotspot. In this study, municipal wastewater was treated by partial nitrification via sequencing batch reactor (SBR), and its effluent and mature landfill leachate were then mixed into an up-flow anaerobic sludge blanket (UASB) for simultaneous anammox and partial denitrification reaction. Through partial nitrification, a high nitrite accumulation rate (93.0 ± 3.8 %) was achieved by low dissolved oxygen (0.5–1.6 mg/L) and controlled aerobic time (3.5 h) in SBR. The UASB system was responsible for 78.8 ± 2.1 % nitrogen removal of the entire system with a hydraulic reaction time (HRT) of 3.8 h, accompanied by the anammox contribution up to 89.4 ± 6.0 %. The overall partial nitrification-simultaneous anammox and partial denitrification (PN-SAPD) system was controlled at a total COD/TIN of 2.8 ± 0.3 and a total HRT of only 10.2 h, achieving the nitrogen removal efficiency and effluent TIN were 95.2 ± 2.2 % and 3.4 ± 1.5 mg/L, respectively. The qPCR results showed functional genes (hzsA(B), hdh) associated with anaerobic ammonia-oxidizing bacteria (AnAOB), whose high gene copy abundance and transcription expression ensured the removal of major nitrogen from municipal wastewater and mature landfill leachate. 16S amplicon sequencing showed that the Ca. Brocadia (9.72–12.6 %) was further enrichment after sodium acetate was added, and the transcription expression of Thauera (0.5–7.0 %) caused nitrate to nitrite. The high abundance of related enzymes (hao, hzs, hdh, narGHI) involved in anammox and partial denitrification processes were found in the macrogenomic sequencing, and only Ca. Brocadia was involved in multi-pathway nitrogen metabolism in AnAOB. Based on the efficient nitrogen removal by AnAOB and denitrifying bacteria, this modified PN-SAPD process provides a new option for the co-treatment of mature landfill leachate in municipal wastewater treatment plants.

Development of partial denitrification process in upflow-anaerobic sludge blanket and effect of electric field on partial denitrification performance

Partial denitrification (PD) is an alternative to providing NO2– for the anaerobic ammonium oxidation (anammox) process. In this study, three upflow anaerobic sludge blankets (UASB) were used to investigate the effect of an external electric field on PD performance. The results indicated that the maximum nitrite transformation ratio (NTR) reached 76.3 %, with an average NTR of 54.1 %, in the presence of external electric field, whereas the average NTR of the control was only 49.8 %. The fitted maximum specific nitrate reduction rates of PD1, PD2, and PD3 were 83.7, 90.5, and 92.3 mg N g−1VSS h−1, respectively, according to the Haldane model analysis. Microbial community analysis demonstrated that the abundance of Thauera, Comamonas, and Accumulibacter increased with electric assistance. In summary, UASB reactor with electrodes set in the upper region was most feasible for the stable PD process, providing an alternative for developing a coupled PD-anammox process.

Municipal wastewater driven partial-denitrification (PD) aggravated nitrous oxide (N2O) production

Partial Denitrification (PD) provides a new route for nitrate-rich wastewater treatment by combining with energy-efficient anammox. This process was reported to hardly produce N2O despite high nitrite accumulation using acetate as carbon source. This study, however, demonstrated significant N2O would be produced when feeding with real municipal wastewater, the NTR-N2O (NO3−-to-N2O transformation ratio) reached as high as 10%. The supply of feed with a high COD/NO3−-N ratio was unable to alleviate N2O production but was severely aggravated after nitrate was depleted. Excess carbon source resulted in accumulated nitrite further being reduced with NTR-N2O largely improved to 63.4%. Besides, it was observed that acid pH inhibited PD activity and resulted in N2O production significantly increased with the NTR-N2O of 39.9% at an initial pH of 5.5. The increase of pH to alkaline conditions cannot mitigate N2O production. A low feeding proportion of real municipal wastewater for low nitrate wastewater treatment can drastically lower N2O production. The NTR-N2O was only 0.1% at a nitrate concentration of 10 mgN/L (10%), much lower than that at 30 mgN/L with a 30% feeding proportion of municipal wastewater (9.4%). Complex organics and ammonium present in municipal wastewater were supposed to be the primary reasons for the high N2O production. Overall, this work offered a new insight into N2O production during the PD process using the complex organics from wastewater itself.

Responses of partial denitrification process to long-term perfluorooctane sulfonic acid stress: Performance, microbial community and functional genes

This study investigated the changes in reactor performances, microbial communities and functional genes in partial denitrification (PD) process under elevated perfluorooctane sulfonic acid (PFOS) stress. The results showed that PFOS significantly (P ≤ 0.001) decreased NO-2-N accumulation rate from 71.15 ± 1.83 % to 65.96 ± 3.97 % and increased total nitrogen removal efficiency from 25.49 ± 1.67 % to 30.90 ± 4.97 %, which indicated that PFOS addition hindered the PD process but promoted the total nitrogen removal. Excitation emission matrix fluorescence spectroscopy and fourier transform infrared spectroscopy suggested significant changes in fluorescence characteristics and functional groups of extracellular polymeric substances, especially, PFOS induced the considerable synthesis of protein-like substances. In addition, high-throughput sequencing revealed that the abundance of genus Thauera, which is responsible for the NO-2-N accumulation, was significantly decreased from 35.98 % to 26.00 % under PFOS stress. While the abundances of common denitrifying bacteria, such as genus Denitratisoma, SM1A02 and OLB8, increased after PFOS exposure, which was consistent with the changes of nitrogen transition. Moreover, PICRUSt showed that the abundance of NO-3-N reductase genes (napA) significantly (P ≤ 0.001) downregulated from 0.02 % to 0.01 %, while NO-2-N reductase genes (nirK) significantly (P ≤ 0.001) upregulated by 1.97 folds after PFOS addition. These results showed that PFOS stress had a profound influence on microbial structure and the functional genes. Overall, this study demonstrated the response mechanism of PD system to long-term PFOS stress and made some contributions to the improvement in the control strategy of wastewater treatment plants (WWTPs).

Deciphering the partial denitrification function of companion bacteria in mixotrophic anammox systems under different carbon/nitrogen ratios

Although partial denitrification coupled with the anammox process has received considerable attention, the partial denitrification function of companion bacteria in mixotrophic anammox systems remains experimentally investigated. Herein, a partial denitrification process was realized in mixotrophic anammox systems with a COD/TN ratio of 0.75. Furthermore, the experimental verification results revealed that nitrate and ammonia were eliminated concurrently, showing that partial denitrifying bacteria were present in the mixotrophic anammox systems. Denitratisoma and Norank Xanthobacteraceae were the dominant genera of companion bacteria containing functional genes related to partial denitrification. Especially, Denitratisoma was a companion bifunctional bacterium for Candidatus Kuenenia shifted between complete and partial denitrification at different COD/TN ratios. Moreover, nir gene abundance in the mixotrophic anammox system increased by 135.4 % compared to that in the autotrophic anammox system. In contrast, nor gene abundance decreased by 28.00 %, indicating that NO might be used as a medium for cross-feeding anammox and partial denitrification. This study provides a more thorough understanding of companion bacteria in mixotrophic anammox systems.

Multi-omics analysis revealed the selective enrichment of partial denitrifying bacteria for the stable coupling of partial-denitrification and anammox process under the influence of low strength magnetic field

The microbial consortium involving anaerobic ammonium oxidation (anammox) and partial denitrification (PD), known as PD-anammox, is an emerging energy-efficient and lower carbon nitrogen removal process from wastewater. However, maintaining a stable PD process by locking nitrate reduction until nitrite was challenging. This study established the first stable connection of anammox with constant nitrite generation by PD bacteria under a low-strength (1.3 mT) magnetic field (MF). When the nitrogen loading rate was 1.81 kg-N/m3/d, the nitrogen removal efficiency of the control reactor (R1) was 75%, lower than that of the experimental reactor (R2), which was 85%. The expression of Thauera and Zoogloea, potential PD bacteria was substantially lower in R1 (5.75% and 1.21%, respectively) than in R2 (10.25 and 6.61%, respectively), according to a meta-transcriptomic analysis. At the same time, the mRNA expression of anammox genera Candidatus Brocadia and Candidatus Kuenenia was 33.53% and 3.83% in R1 and 22.86% and 1.87% in R2. Moreover, carbon and nitrogen metabolism pathways were more abundant under the influence of low-strength MF. The selective enrichment of PD bacteria can be attributed to the increased expression of carbon metabolic pathways like the citrate cycle, glycolysis/gluconeogenesis, and pyruvate metabolism. Interestingly, the control reactor was dominated by a hydroxylamine-dependent anammox process while a low-strength MF-enhanced nitric-oxide-dependent anammox process. For successful anammox-centered nitrogen removal from wastewater, this study demonstrated that low-strength MF is a convenient and applicable technique to lock the nitrate reduction until nitrite.

Hydroxyapatite (HAP) formation in acetate-driven partial denitrification process: Enhancing sludge granulation and phosphorus removal

Partial denitrification/anammox (PD/A) processes have emerged as a promising technology for efficient nitrogen removal from wastewater. However, these processes fail to remove phosphorus (P), a key pollutant that contributes to water eutrophication. To address this issue, the potential of inducing hydroxyapatite (HAP) precipitation in PD processes to achieve simultaneous P removal was investigated for the first time. Specifically, three SBRs (R1-R3) for PD were operated with adding varying concentrations of external Ca (30, 60, and 120 mg/L, respectively). Results demonstrated significant P reduction in all three SBRs, particularly in R3 with high Ca, which achieved an 80 % removal efficiency. Notably, sludge granulation was observed during operation, with the granule size in R3 with high Ca reaching 906.1 μm during the stable period, exceeding those in R2 (788.7 μm) and R1 (707.1 μm). This led to good settle ability of the PD sludge, as demonstrated by the lowest SVI5 (20 mL/g MLSS). Moreover, the decrease in the MLVSS/MLSS ratio suggested that the inorganic content accumulated, as observed by confocal laser scanning microscopy in the interior of the granules. Elemental composition analysis suggested that PD granules contained high P and Ca, while the X-ray diffraction (XRD) results confirmed the formation of HAP. Overall, this study demonstrated that PD-HAP coupled granular sludge process has potential as a robust and efficient method for nitrite production, as well as effective P removal and recovery, thereby advancing the application of anammox processes in wastewater treatment.

Insights into the Responses of the Partial Denitrification Process to Elevated Perfluorooctanoic Acid Stress: Performance, EPS Characteristic and Microbial Community

This study aimed at investigating the potential impacts of perfluorooctanoic acid (PFOA) exposure on the partial denitrification (PD) system. Our results indicated that nitrite accumulation rates were significantly decreased to 67.94 ± 1.25%–69.52 ± 3.13% after long-term PFOA exposure (0.5–20 mg/L), while the nitrate transformation ratio was slightly impacted. The PFOA removal efficiency gradually decreased from 67.42 ± 3.39% to 6.56 ± 5.25% with an increasing PFOA dosage, indicating that the main PFOA removal pathway was biosorption. The average EPS contents increased by two folds, which suggested that exposure to PFOA significantly stimulated EPS secretion. Excitation emission matrix analysis revealed that PFOA exposure promoted the secretion of tryptophan protein-like, humic acid-like, and aromatic protein II-like substances, which may act as a protective barrier against PFOA toxicity. Moreover, significant changes in characteristic peaks after PFOA exposure were shown as indicated by Fourier transform infrared spectroscopy. High-throughput sequencing suggested that PFOA significantly decreased bacterial richness and increased evenness, indicating that toxicity effects of PFOA were more pronounced for abundant species (e.g., Thauera) than rare species. Thauera was the most dominant genus responsible for nitrite accumulation, whose abundance significantly decreased from 35.99 ± 2.67% to 18.60 ± 2.18% after PFOA exposure. In comparison, the abundances of common denitrifiers, such as Denitratisoma, Bdellovibrio, and OLB8, significantly increased, suggesting that these genera were potential PFOA-resistant bacteria. This study presents new insights into the effect of PFOA on a PD system.

Revealing the metabolic characteristics and nitrite accumulation mechanism of glycerol-driven partial denitrification process by batch tests and untargeted metabolomic analysis

Glycerol has been proven as an effective carbon source for partial denitrification (PD), while the underlying mechanism of the nitrite accumulation remains unknown. In this study, the nitrite accumulation characteristics was firstly investigated by batch tests. Denitrifying bacteria exhibited high nitrate to nitrite ratio (NTR) (84.6%) and nitrite accumulation rate (NAR) (30.23 mg N h−1 gMLVSS−1) in a typical PD process with sufficient glycerol supply. But when glycerol was exhausted, denitrifying bacteria entered endogenous PD process driven by polyhydroxyalkanoates (PHA) and glycogen, and the NTR and NAR significantly decreased to 63.9% and 1.74 mg N h−1 gMLVSS−1, respectively. More analysis found that keeping the endogenous carbon source at a relatively high level before PD reaction could further improved the NTR up to 99.7%. Metabolomic analysis showed that most required amino acids were synthesized in nitrite accumulation phase with active RNA metabolism. While in the subsequent nitrite reduction phase with limited glycerol, microbes tended to synthesize those amino acids with lower energetic cost, and activities related to DNA metabolism was more active in this phase. Moreover, reactive oxygen species generated in the glycerol degradation were deduced to be the key factor that induced the efficient nitrite accumulation. This study provided a comprehensive insight into the mechanism of glycerol-driven PD process.

Response of nitrite accumulation to elevated C/NO– 3-N ratio during partial denitrification process: Insights of extracellular polymeric substance, microbial community and metabolic function

This study investigated the response of nitrite accumulation to elevated COD/NO3–-N ratio (C/N) during partial denitrification (PD). Results indicated nitrite was gradually accumulated and remained stable (C/N = 1.5 ∼ 3.0), while that rapidly declined after reaching the peak (C/N = 4.0 ∼ 5.0). The polysaccharide (PS) and protein (PN) content of tightly-bound extracellular polymeric substances (TB-EPS) reached the maximum at C/N of 2.5 ∼ 3.0, which might be stimulated by high level of nitrite. Illumina MiSeq sequencing showed Thauera and OLB8 were dominated denitrifying genera at C/N of 1.5 ∼ 3.0, while Thauera was further enriched with fading OLB8 at C/N of 4.0 ∼ 5.0. Meanwhile, the highly-enriched Thauera might enhance the activity of nitrite reductase (nirK) promoting further nitrite reduction. Redundancy analysis (RDA) showed positive correlations between nitrite production and PN content of TB-EPS, denitrifying bacteria (Thauera and OLB8) and nitrate reductases (narG/H/I) in low C/N. Finally, their synergistic effects for driving nitrite accumulation were comprehensively elucidated.

Extending reaction duration has minor influence on nitrite production in partial-denitrification process

Partial denitrification (PD) is another important pathway producing nitrite for anammox, however, whether its performance is affected by overlong reaction time, a situation that often takes place is still unknown. Three sequencing batch reactors were operated for PD to evaluate this factor on nitrite production. Results indicated effluent nitrite was very close despite reaction time even extending to four times longer than control (i.e., nitrate-to-nitrite transformation ratio (NTR) of 94.4%-89.8%). Meanwhile, it was found PD could recover to the normal after suffering from high organics shocking. Cycle studies suggested produced nitrite would not be further reduced with prolonged time, as indicated by changing trend of pH and alkalinity. Microbial analysis revealed PD functional bacteria, Thauera, slightly decreased with prolonged reaction, while it was always predominated. Taken together, this study indicated overlong reaction time had minor influence on PD, demonstrating its robustness with great technological superiority in supplying nitrite for anammox.

Exposure to Cr(VI) affects partial denitrification process-nitrite accumulation, EPS characteristic and microbial community assembly

This study was aimed at assessing the effects of elevated Cr(VI) stresses on partial denitrification (PD) process. The results showed that 0.5–5 mg/L Cr(VI) exposure significantly decreased nitrite accumulation rate, while COD removal efficiencies increased due to the further conversion of NO2--N to N2. This suggested that Cr(VI) exposure shifted the biological process from partial denitrification to complete denitrification. The Cr(VI) removal efficiency decreased from 37.75 ± 15.97% to 10.48 ± 3.78% with increasing Cr(VI) concentration. Excitation-emission matrix fluorescence spectroscopy showed that Cr(VI) exposure significantly stimulated the synthesis of tryptophan protein-like, aromatic protein-like and humic acid-like substances in extracellular polymeric substances (EPS). In addition, significant changes in functional groups of EPS were found as revealed by Fourier transform infrared spectroscopy. Particularly, there was a considerable decrease in the α-Helix from 31.36% to 0% and a major increase in the antiparallel β-Sheet from 4.81% to 9.03% with increasing Cr(VI) stresses. The biodiversity decreased significantly under Cr(VI) stresses, and the microbial community structures gradually shifted with increasing Cr(VI) dosage. Sloan model revealed that the relative importance of deterministic processes on the community assembly increased after Cr(VI) exposure, indicating that Cr(VI)-induced selective processes have a profound influence on microbial community of PD process. Network analysis revealed that Cr(VI) exposure significantly altered species-species associations, topological features of individual OTU, and keystone species, implying that substantial changes occurred in bacterial ecology after exposure to Cr(VI).