Ammonia Oxidation Rate Research Articles

Optimizing heterotrophic nitrification process: The significance of demand-driven aeration and organic matter concentration

Heterotrophic nitrification and aerobic denitrification (HNAD) sludge were successfully acclimated. The effects of organics and dissolved oxygen (DO) on nitrogen and phosphorus removal by the HNAD sludge were investigated. The nitrogen can be heterotrophically nitrified and denitrified in the sludge at a DO of 6 mg/L. The TOC/N (total organic carbon to nitrogen) ratio of 3 was found to result in removal efficiencies of over 88% for nitrogen and 99% for phosphorus. The use of demand-driven aeration with a TOC/N ratio of 1.7 improved nitrogen and phosphorus removal from 35.68% and 48.17% to 68% and 93%, respectively. The kinetics analysis generated an empirical formula, Ammonia oxidation rate = 0.08917·(TOC·Ammonia)0.329·Biomass0.342. The nitrogen, carbon, glycogen, and poly-β-hydroxybutyric acid (PHB) metabolism pathways of HNAD sludge were constructed using the Kyoto Encyclopedia of Genes and Genomes (KEGG). The findings suggest that heterotrophic nitrification precedes aerobic denitrification, glycogen synthesis, and PHB synthesis.

Modeling the kinetics of perfluorooctanoic and perfluorooctane sulfonic acid biodegradation by Acidimicrobium sp. Strain A6 during the feammox process

Per- and polyfluoroalkyl substances (PFAS) are emerging contaminants of concern due to their health effects and persistence in the environment. Although perfluoroalkyl acids (PFAAs), such as perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) are very difficult to biodegrade because they are completely saturated with fluorine, it has recently been shown that Acidimicrobium sp. A6 (A6), which oxidizes ammonium under iron reducing conditions (Feammox process), can defluorinate PFAAs. A kinetic model was developed and tested in this study using results from previously published laboratory experiments, augmented with results from additional incubations, to couple the Feammox process to PFAS defluorination. The experimental results show higher Feammox activity and PFAS degradation in the A6 enrichment cultures than in the highly enriched A6 cultures. The coupled experimental and modeling results show that the PFAS defluorination rate is proportional to the rate of ammonium oxidation. The ammonium oxidation rate and the defluorination rate increase monotonically, but not linearly, with increasing A6 biomass. Given that different experiments had different level of Feammox activity, the parameters required to simulate the Feammox varied between A6 cultures. Nonetheless, the kinetic model was able to simulate an anaerobic incubation system and show that PFAS defluorination is proportional to the Feammox activity.

Mechanisms Driving the Distribution and Activity of Mineralization and Nitrification in the Reservoir Riparian Zone.

The riparian zone ecosystems have greater energy flow and elemental cycling than adjacent terrestrial and aquatic ecosystems. Mineralization and nitrification are important initiating processes in the nitrogen cycle, but their distribution and activity under different environmental conditions in the riparian zone and the driving mechanisms are still not clear. We investigated the effects of environmental and microbial factors on mineralization and nitrification activities by analyzing the community of alkaline (apr) and neutral (npr) metallopeptidase, ammonia-oxidizing archaea (AOA), and bacteria (AOB) in soils and sediments under different land-use types in the riparian zone of Miyun Reservoir, as well as measuring potential nitrogen mineralization and ammonia oxidation rates (AOR). The results showed that the mineralization and nitrification activities of soils were greater than those of sediments. AOA and AOB dominate the ammonia oxidation activity of soil and sediment, respectively. NH4+ content was a key factor influencing the ecological niche differentiation between AOA and AOB. The high carbon and nitrogen content of the woodland significantly increased mineralization and nitrification activity. Microbial communities were significantly clustered in the woodland. The land-use type, not the flooding condition, determined the distribution of microbial community structure. The diversity of npr was significantly correlated with potential N mineralization rates, while the transcript abundance of AOA was significantly correlated with ammonia oxidation rates. Our study suggests that environmental changes regulate the distribution and activity of mineralization and nitrification processes in the reservoir riparian zone by affecting the transcript abundance, diversity and community structure of the microbial functional genes.

Comparative assessment of energy generation from ammonia oxidation by different functional bacterial communities

Bioelectrochemical ammonia oxidation (BEAO) in a microbial fuel cell (MFC) is a recently discovered process that has the potential to reduce energy consumption in wastewater treatment. However, level of energy and limiting factors of this process in different microbial groups are not fully understood. This study comparatively investigated the BEAO in wastewater treatment by MFCs enriched with different functional groups of bacteria (confirmed by 16S rRNA gene sequencing): electroactive bacteria (EAB), ammonia oxidizing bacteria (AOB), and anammox bacteria (AnAOB). Ammonia oxidation rates of 0.066, 0.083 and 0.082 g NH4+-N L−1 d−1 were achieved by biofilms enriched with EAB, AOB, and AnAOB, respectively. With influent 444 ± 65 mg NH4+-N d−1, nitrite accumulation between 84 and 105 mg N d−1 was observed independently of the biofilm type. The AnAOB-enriched biofilm released electrons at higher potential energy levels (anode potential of 0.253 V vs. SHE) but had high internal resistance (Rint) of 299 Ω, which limits its power density (0.2 W m−3). For AnAOB enriched biofilm, accumulation of nitrite was a limiting factor for power output by allowing conventional anammox activity without current generation. AOB enriched biofilm had Rint of 18 ± 1 Ω and yielded power density of up to 1.4 W m−3. The activity of the AOB-enriched biofilm was not dependent on the accumulation of dissolved oxygen and achieved 1.5 fold higher coulombic efficiency when sulfate was not available. The EAB-enriched biofilm adapted to oxidize ammonia without organic carbon, with Rint of 19 ± 1 Ω and achieved the highest power density of 11 W m−3. Based on lab-scale experiments (scaling-up factors not considered) energy savings of up to 7 % (AnAOB), 44 % (AOB) and 475 % (EAB) (positive energy balance), compared to conventional nitrification, are projected from the applications of BEAO in wastewater treatment plants.

Effect of free nitrous acid on nitritation process: Microbial community, inhibitory kinetics, and functional biomarker

This work comprehensively deciphered the effect of free nitrous acid (FNA) on the microbial community, inhibitory kinetics, and nitrifiers in nitritation process. Nitritation was first successfully achieved through selective inhibition of free ammonia (FA) on nitrite oxidizers (NOB). Then, batch tests clearly showed that FNA significantly inhibits the ammonia oxidation rate (rsu) and the growth rate (μ) of ammonia oxidizers (AOB), which was well described by the Hellinga model (KI = 0.222 mg·L−1). The structural equation model indicated that FNA was significantly and negatively associated with rsu, μ, Nitrosomonas, Commamons, Nitrospira, and Nitrotoga and positively correlated with Paracoccus. Furthermore, Nitrosomonas significantly drove the ammonia utilization and growth of AOB and was identified as the most important functional biomarker indicating the nitritation in response to FNA levels using random forest model. This study provides helpful information on the kinetics of the mechanism underlying the FNA inhibition on nitrification.

Study of effect of various temperatures on the abundance of ammonia oxidizing archaea and bacteria

Temperature plays significant role in the oxidation of ammonia in filtration units of recirculating aquaculture system. The impact of temperature on the abundance of ammonia oxidizing archaea and bacteria, and the expression of ammonia oxidizing gene (amoA) at specific temperature was evaluated. The broken earthen pot pieces used as filter bed materials of recirculating system, showing the presence of microorganisms were introduced in glass containers (5 pieces/5l) filled with synthetic wastewater and exposed to four different temperatures of 10, 20, 30 and 40°C for 40 days. The ammonia oxidation rate was minimum at 10°C. In 20, 30 and 40°C treatments, 99% ammonia was reduced on day-18, 8 and 18, respectively compared to the initial day. Fresh ammonium chloride (2 mM) was added twice to maintain the ammonia concentration in all treatments, except 10°C one. Nitrite-N level was < 1 mg/l at 10°C. The level was highest on day-22 at 20° and 40°C and on day-12 at 30°C. The nitrification was 10 days delayed at 20°C and 40°C compared to 30°C treatment. Concentration of nitrate-N was lowest at 10°C. Highest concentration of nitrate-N was observed on day-40 at 20°C and 40°C and day-26 at 30°C. Highest copy number of bacterial amoA was recorded at 30°C (2.59×107) followed by 20°C (4.08×106), 40°C (1.45×106) and 10°C (5.664×103). Archaeal amoA was highest at 30°C (7.47×103) followed by 40°C (2.98×102) and 20°C (46.8) treatments. Hence it may be concluded that 30°C temperature was optimum for the efficient and faster oxidation of ammonia in the present recirculating system.

Activated sludge vs. biofilm – effect of temperature on ammonia and nitrite oxidation rate in the hybrid reactor

Activated sludge vs. biofilm – effect of temperature on ammonia and nitrite oxidation rate in the hybrid reactor

New insights into exogenous N-acyl-homoserine lactone manipulation in biological nitrogen removal system against ZnO nanoparticle shock

The effects and mechanisms of three N-acyl-homoserine lactones (AHLs) (C4-HSL, C6-HSL, and C10-HSL) on responses of biological nitrogen removal (BNR) systems to zinc oxide nanoparticle (NP) shock were investigated. All three AHLs improved the NP-impaired ammonia oxidation rates by up to 50.0 % but inhibited the denitrification process via regulating nitrogen metabolism-related enzyme activities. C4-HSL accelerated the catalase activity by 13.2 %, while C6-HSL and C10-HSL promoted the superoxide dismutase activity by 26.6 % and 18.4 %, respectively, to reduce reactive oxygen species levels. Besides, the enhancements of tryptophan protein and humic acid levels in tightly-bound extracellular polymeric substance by AHLs were vital for NP toxicity attenuation. The metabonomic analysis demonstrated that all three AHLs up-regulated the levels of lipid- and antioxidation-related metabolites to advance the system’s resistance to NP shock. The “dual character” of AHLs emphasized the concernment of legitimately employing AHLs to alleviate NP stress for BNR systems.

Influence of supplementary carbon on reducing the hydraulic retention time in microalgae-bacteria (MaB) treatment of municipal wastewater

Real municipal wastewater was treated using microalgae and bacteria (MaB) flocs with the goal to achieve the European standards (EUS91) in less than 2 days of hydraulic retention time (HRT). The pH in the control series (CS) was allowed to increase freely. In the inorganic carbon supplement (ICS) series, the pH was maintained below 8.0 by means of CO2 injection. The presence of CO2 regulated the nitrogen profiles, but the total removal did not improve. Nitrification was stronger and advanced more when CO2 was applied during 2 days of HRT with an average ammonium oxidation rate of 40.5 ± 4.9 %. The residual phosphate and nitrogen oxides were assimilated on the third day of treatment. Balancing the organic carbon (COD:NH4–N ratio of 15.9) suppressed the autotrophic activities (nitrification) and improved the biomass assimilation significantly. Batch experiments with balanced organic carbon could remove nitrogen and phosphorus with rates of 43.1 ± 5.1 mg N L−1 d−1 (88.3 ± 13.9 %) and 6.21 ± 0.07 mg PO4–P L−1 d−1 (98.2 ± 1.6 %) after 28 hours. Our results show that balanced MaB culture can satisfy the EUS91 limits within approximately 24 hours of treatment.

Evaluation of the stability of shortcut nitrification-denitrification process based on online specific oxygen uptake rate monitoring

Shortcut nitrification-denitrification (SCND) is widely concerned because of its low energy consumption and high nitrogen removal efficiency. However, the current difficulty lies in the stable maintenance of SCND performance, which leads to the challenge of large-scale application of this new denitrification technology. In this study, the nitrogen removal pathway from complete nitrification-denitrification (CND) to SCND was rapidly realized under high free ammonia (FA), high pH and low dissolved oxygen (DO) conditions. The variations of specific oxygen uptake rate (SOUR) of activated sludge in both processes were investigated by an online SOUR monitoring device. Different curves of SOUR from CND to SCND process were observed, and the ammonia peak obtained based on SOUR monitoring could be used to control aeration time accurately in SCND process. Accordingly, the SOUR ratio of ammonia oxidizing bacteria (AOB) to nitrite oxidizing bacteria (NOB) (SOURAOB/SOURNOB) was increased from 1.40 to 2.93. 16S rRNA Miseq high throughput sequencing revealed the dynamics of AOB and NOB, and the ratio of relative abundance (AOB/NOB) was increased from 1.03 to 3.12. Besides, SOURAOB/SOURNOB displayed significant correlations to ammonia removal rate (P < 0.05), ammonia oxidation rate / nitrite oxidation rate (P < 0.05), nitrite accumulation rate (P < 0.05) and the relative abundance of AOB/NOB (P < 0.05). Thus, a strategy for evaluation the SCND process stability based on online SOUR monitoring is proposed, which provides a theoretical basis for optimizing the SCND performance.

Efficient partial nitrification with hybrid nitrifying granular sludge based on a simultaneous fill/draw SBR mode

In this study, a simultaneous fill/draw SBR was applied to investigate the feasibility of partial nitrification process with inoculation of matured aerobic granular sludge. The system operated stably over 120 days with the relatively high ammonium removal efficiency (≥ 98.83%) and nitrite accumulation rate (≥ 89.60%). Moreover, a hybrid flocs/granules system was formed stably after long-term operation. The nitrite-oxidizing bacteria (NOB) was suppressed effectively because of the combined effect of simultaneous fill/draw mode and intermittent aeration conditions. Furthermore, batch tests were separately tested with isolated granules (> 200 μm) and flocs (< 200 μm), showing that the specific ammonia oxidation rate of granules and flocs were 15.94 ± 2.85 and 66.77 ± 0.83 mg N/(g MLSS·h), respectively. Correspondingly, the abundance of Nitrosomonas as a typical AOB in granules (6.24%) and flocs (11.94%) was obtained via the microbial diversity analysis, while NOB was almost hardly detected in granules and flocs.

Iron-containing carriers stimulate nitrogen conversions in one-stage reactors treating N-rich digester supernatant

Iron incorporation in biomass carriers (0%, 1%, 5%, and 10% of carrier weight) at nitrogen loading rates (NLR) from 0.08 to 0.30 g/(g MLVSS·d) was tested during the treatment of nitrogen-rich digester supernatant in one-stage reactors, in which hybrid biomass (suspended/attached) was inhabited by ammonia-oxidizing bacteria, Anammox bacteria, and denitrifiers. The Anammox process dominated at the highest NLR and increased the volumetric nitrogen removal rate to about 500 g/(m3·d). Total microbial composition was most strongly affected by NLR, however, Anammox bacteria abundance was also affected by the iron content. These bacteria predominated in reactors with 5%-iron carriers, which ensured the highest nitrogen removal efficiency (up to 75%) and kinetics: the average ammonium removal rate was at least 40% higher than in the reactor without iron. The presence of 10%-iron carriers in the reactor inhibited the ammonium oxidation rate; an increase in ammonium removal rate with increasing initial ammonium concentration was the lowest. The contribution of Anammox to nitrogen removal was demonstrated by the simultaneous increase in biomass heme c content to about 0.1 µmol/mg protein along with increasing iron content to 5% and NLR. The incorporation of iron into the carrier structure allowed for a long-term release of mostly ferrous iron to the supernatant, thus accelerating its treatment.

Controls on the relative abundances and rates of nitrifying microorganisms in the ocean

Abstract. Nitrification controls the oxidation state of bioavailable nitrogen. Distinct clades of chemoautotrophic microorganisms – predominantly ammonia-oxidizing archaea (AOA) and nitrite-oxidizing bacteria (NOB) – regulate the two steps of nitrification in the ocean, but explanations for their observed relative abundances and nitrification rates remain incomplete and their contributions to the global marine carbon cycle via carbon fixation remain unresolved. Using a mechanistic microbial ecosystem model with nitrifying functional types, we derive simple expressions for the controls on AOA and NOB in the deep, oxygenated open ocean. The relative biomass yields, loss rates, and cell quotas of AOA and NOB control their relative abundances, though we do not need to invoke a difference in loss rates to explain the observed relative abundances. The supply of ammonium, not the traits of AOA or NOB, controls the relatively equal ammonia and nitrite oxidation rates at steady state. The relative yields of AOA and NOB alone set their relative bulk carbon fixation rates in the water column. The quantitative relationships are consistent with multiple in situ datasets. In a complex global ecosystem model, nitrification emerges dynamically across diverse ocean environments, and ammonia and nitrite oxidation and their associated carbon fixation rates are decoupled due to physical transport and complex ecological interactions in some environments. Nevertheless, the simple expressions capture global patterns to first order. The model provides a mechanistic upper estimate on global chemoautotrophic carbon fixation of 0.2–0.5 Pg C yr−1, which is on the low end of the wide range of previous estimates. Modeled carbon fixation by AOA (0.2–0.3 Pg C yr−1) exceeds that of NOB (about 0.1 Pg C yr−1) because of the higher biomass yield of AOA. The simple expressions derived here can be used to quantify the biogeochemical impacts of additional metabolic pathways (i.e., mixotrophy) of nitrifying clades and to identify alternative metabolisms fueling carbon fixation in the deep ocean.

Functional biological anode as electron acceptor for direct oxidation of ammonia nitrogen to N2 in an anaerobic single-chamber reactor

A functional biological anode (nonwoven-wrapped functional microbial carbon felt bag) was constructed to realize the ammonia oxidation to nitrogen gas in a single-chamber anaerobic microbial electrochemical system (MES). The applied potential has a significant impact on oxidation products of ammonia. As the applied potential was reduced from +0.8 V to +0.3 V (vs. Ag/AgCl), the ammonia oxidation rate increased from 65.2% to 94.7% and the TN removal rate increased from almost 0 to 88.0% on the bio-anode. At +0.3 V ammonia could be converted to N2 on the bio-anode and hydrogen is generated on the cathode. The zeolite adsorbent added inside the anodic carbon felt bag was also a key factor to accelerate nitrogen removal in the MES. The dominant genera of SWB02 sp., Nitrospira sp. and Bryobacter sp. played a crucial role in ammonia oxidation according to 16S rRNA analysis. This paper provides a new idea for NH4+-N removal and hydrogen production from wastewater.

Long-term N-addition alters the community structure of functionally important N-cycling soil microorganisms across global grasslands

Anthropogenic nitrogen (N) input is known to alter the soil microbiome, but how N enrichment influences the abundance, alpha-diversity and community structure of N-cycling functional microbial communities in grasslands remains poorly understood. Here, we collected soils from plant communities subjected to up to 9 years of annual N-addition (10 g N m−2 per year using urea as a N-source) and from unfertilized plots (control) in 30 grasslands worldwide spanning a large range of climatic and soil conditions. We focused on three key microbial groups responsible for two essential processes of the global N cycle: N2 fixation (soil diazotrophs) and nitrification (AOA: ammonia-oxidizing archaea and AOB: ammonia-oxidizing bacteria). We targeted soil diazotrophs, AOA and AOB using Illumina MiSeq sequencing and measured the abundance (gene copy numbers) using quantitative PCR. N-addition shifted the structure of the diazotrophic communities, although their alpha-diversity and abundance were not affected. AOA and AOB responded differently to N-addition. The abundance and alpha-diversity of AOB increased, and their community structure shifted with N-addition. In contrast, AOA were not affected by N-addition. AOA abundance outnumbered AOB in control plots under conditions of low N availability, whereas N-addition favoured copiotrophic AOB. Overall, N-addition showed a low impact on soil diazotrophs and AOA while effects for AOB communities were considerable. These results reveal that long-term N-addition has important ecological implications for key microbial groups involved in two critical soil N-cycling processes. Increased AOB abundance and community shifts following N-addition may change soil N-cycling, as larger population sizes may promote higher rates of ammonia oxidation and subsequently increase N loss via gaseous and soil N-leaching. These findings bring us a step closer to predicting the responses and feedbacks of microbial-mediated N-cycling processes to long-term anthropogenic N-addition in grasslands.

Differential bacterial ammonia oxidation patterns in soil particles from two contrasting forests: The importance of interfacial interactions

Soil inorganic nitrogen (N) deficiency limits forest productivity in subalpine regions. The traditional view holds that subalpine coniferous forests are characterized by lower soil inorganic N availability, and such differences are greatly affected by the diversity and abundance of ammonia-oxidizing bacteria. It is known that interfacial interactions between bacteria and solid particles in agricultural and aquatic ecosystems can alter bacterial metabolic activity and biogeochemical processes. However, whether bacteria-soil interfacial interactions induce different ammonia oxidation processes in distinct soil particles remain unknown. Herein, we collected soil particles of different sizes from two contrasting forest sites (a spruce-dominated natural forest and spruce plantation) and examined the effect of soil-Nitrosomonas europaea (a model ammonia-oxidizing bacteria) interfacial interactions on the oxidation of ammonium (NH4+-N) using batch adsorption and semipermeable membrane experiments. The metabolic activity of bacteria was also examined using an isothermal microcalorimetric method. Upon adsorption on soil particles, the ammonia oxidation rate and bacterial metabolism activity were slower than those of free bacteria. The inhibition effect ceased when the bacteria were physically isolated from the soil particles by a membrane, indicating the importance of interfacial interactions in the ammonia oxidation process. Additionally, clay particles were more effective than silts and sands in binding cells and greatly reduced the ammonia oxidation reaction. Across the two forests, natural forest soils exhibited a higher bacterial adsorption capacity but lower bacterial metabolic activity, resulting in a fewer loss of NH4+-N. Our results suggested that stronger bacteria-soil interfacial interactions (especially for clay fractions) were responsible for maintaining high soil inorganic N availability in forests, highlighting the need to incorporate interfacial interactions into biogeochemical models of N cycling in forest ecosystems.

Shifts of ammonia-oxidation process along salinity gradient in an estuarine wetland

Ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) play an important role in nitrogen (N) cycling of the estuarine sediments. However, the shifts of potential ammonia oxidation rates (PARs) of AOA and AOB along different salinity gradients are not well understood. Here, salinity-treated (0, 5, 15, 25 and 35 ‰) incubation experiments of sediment from the Pearl River Estuary were performed to explore the influence of salinity on ammonia oxidation process, including the PARs and gene abundances of AOA and AOB. Significant shifts of PARs and amoA gene abundances of ammonia oxidation process along salinity gradient were observed during the incubation (0–60 d). After a long-term incubation, both PARtotal (4.61 ± 0.93 μg N g−1 d−1) and PARaob (3.05 ± 1.04 μg N g−1 d−1) reached their maximum values at 25 ‰ salinity, and PARaoa (1.96 ± 0.10 μg N g−1 d−1) reached its maximum values at 35 ‰ salinity, which showed that PARs had higher values under high salinity conditions. AOB amoA gene was more abundant than AOA under moderate and high salinity, and only a significant linear correlation was observed between PARaob and AOB abundances. In addition, the ammonia oxidation activities showed a wide range of salinity tolerance, which is worthy of further study on community structure and transcript richness. Our study illustrates that salinity plays a key role in regulating the ammonia oxidation process in estuarine sediments, and has guiding significance for N fixation and N loss associated with saltwater incursion.

Responses of various carbon to nitrogen ratios to microbial communities, kinetics, and nitrogen metabolic pathways in aerobic granular sludge reactor

The role of different ammonia concentrations (mg N/L) (of 100 (carbon to nitrogen ratio (C/N) = 12; Stage I), 200 (C/N = 6; Stage II), 400 (C/N = 3; Stage III) and 200 (C/N = 6; Stage IV)) in nitrogen metabolic pathways, microbial community, and specific microbial activity were investigated in an aerobic granular sludge reactor. Heterotrophic ammonia oxidizing bacteria (HAOB) showed higher ammonia oxidation rates (AORs) than autotrophic ammonia oxidizing bacteria (AAOB) at higher C/N conditions (Stages I and II). Paracoccus was the dominant HAOB. AAOB, with only 0.2–0.3 % in relative abundance, showed 2.7-fold higher AORs than HAOB at elevated ammonia and free ammonia (FA) concentrations with C/N at 3. Nitrosomonas and a genus in Nitrosomondaceae family were the major AAOB. This study proposed that FA inhibition on heterotrophic bacteria might be the mechanism that contributes to the development of the autotrophic ammonia oxidation pathway and enrichment of AAOB.

An enriched ammonia-oxidizing microbiota enables high removal efficiency of ammonia in antibiotic production wastewater

High ammonia concentration hinders the efficient treatment of antibiotic production wastewater (APW). Developing effective ammonia oxidation wastewater treatment strategies is an ideal approach for facilitating APW treatment. Compared with traditional nitrification strategies, the partial nitrification process is more eco-friendly, less energy-intensive, and less excess sludge. The primary limiting factor of the partial nitrification process is increasing ammonia-oxidizing bacteria (AOB) while decreasing nitrite-oxidizing bacteria (NOB). In this study, an efficient AOB microbiota (named AF2) was obtained via enrichment of an aerobic activated sludge (AS0) collected from a pharmaceutical wastewater treatment plant. After a 52-day enrichment of AS0 in 250 mL flasks, the microbiota AE1 with 69.18% Nitrosomonas microorganisms was obtained. Subsequent scaled-up cultivation in a 10 L fermenter led to the AF2 microbiota with 59.22% Nitrosomonas. Low concentration of free ammonia (FA, < 42.01 mg L−1) had a negligible effect on the activity of AF2, and the nitrite-nitrogen accumulation rate (NAR) of AF2 was 98% when FA concentration was 42.01 mg L−1. The specific ammonia oxidation rates (SAORs) at 30 °C and 15 °C were 3.64 kg NH4+-N·kg MLVSS−1·d−1 and 1.43 kg NH4+-N·kg MLVSS−1·d−1 (MLVSS: mixed liquor volatile suspended solids). The SAOR was 0.52 kg NH4+-N·kg MLVSS−1·d−1 when the NaCl concentration was increased from 0 to 20 g L−1, showing that AF2 functioning was stable in a high-level salt environment. The ammonia oxidation performance of AF2 was verified by treating abamectin and lincomycin production wastewater. The NARs of AF2 used for abamectin and lincomycin production wastewater treatment were >90% and the SAORs were 2.39 kg NH4+-N·kg MLVSS−1·d−1 and 0.54 kg NH4+-N·kg MLVSS−1·d−1, respectively, which was higher than the traditional biological denitrification process. In summary, AF2 was effective for APW treatment via enhanced ammonia removal efficiency, demonstrating great potential for future industrial wastewater treatment.

Comammox biogeography subject to anthropogenic interferences along a high-altitude river

The recent discovery of comammox Nitrospira performing complete ammonia oxidation to nitrate has overturned the long-held dogma of two-step nitrification on Earth, yet little is known about the effect of urbanization interference on their distribution. Using gene-centric metagenomics, we provided the first blueprints about comammox community, biogeography, and environmental drivers along a high-elevation (> 2000 m) river flowing through the largest city on the vulnerable Qinghai-Tibetan Plateau. Our study confirmed a wide presence and diversity of yet-uncultured comammox clade B across wet and dry seasons, with average 3.0 and 2.0 times as abundant as clade-A amoA genes in water and sediments, respectively. Species identified from freshwater and drinking water treatment plants dominated the comammox guilds (58∼100%), suggesting this plateau river shared a similar comammox assemblage with the above habitat types. Compared with the urban area harboring more abundant canonical Nitrospira identified in wastewater (average 24%), the upstream suburban reach had a smaller human population but larger proportions of comammox in ammonia-oxidizing prokaryotes (24∼72% of abundances) and Nitrospira sublineages I/II. Higher contents of nitrate and nitrite in water, and antibiotics in water and sediments, may restrain comammox niches in nitrifiers over the urban area. Further random forest analysis revealed that lincosamides and quinolones were the most important antibiotic predictors for the niche differentiations between comammox and canonical nitrifiers in water, while macrolides for those in sediments. Finally, by incubation experiments, we demonstrated higher activity contributions of benthic comammox in the suburban area (36.2∼92.8% of potential ammonia-oxidation rates) than in the urban reach, and that the contribution variation had significant negative relations with macrolides and their major components. Overall, this study highlighted that anthropogenic activities hampered the advantage of riverine complete nitrifiers over the canonical two-step ones.