Response of nitrifiers to gradually increasing pH conditions in a membrane nitrification bioreactor: Microbial dynamics and alkali-resistant mechanism.

Nitrification and nitrifiers are pH-sensitive especially under the alkaline environment in the activated sludge system. However, it is unclear how nitrifiers and nitrification respond to long-term alkaline environment. This study employed a continuous flow membrane nitrification bioreactor to investigate the dynamics of nitrification efficiency and microbial community adaptation under a 320-day alkaline operation. Results showed that activated sludge adapted remarkably to a progressive increase in pH from 7.5 to 10.0, achieving robust nitrification with average ammonia removal efficiencies of 96.6 ± 2.2%. Subsequently, an integrated alkali-resistant mechanism of nitrifiers was proposed. Specifically, under the long-term operation of pH 10.0, certain bacteria secreted enhanced extracellular acidic polysaccharides (i.e., up to 10.95 ± 0.27 mg·g-1 MLVSS in soluble extracellular polymeric substances (EPS)) and acidic organic compounds (e.g., humic acids increased by 1.47-fold in tightly bounded EPS) to neutralize external alkalinity. Moreover, significant enrichments in both the ammonia oxidizing bacteria Nitrosomonas (by 1.3%) and the nitrite oxidizing bacteria Nitrospira (by 5.4%) were observed in a 170-day operation of pH 10.0 condition. Meanwhile, norank_f__JG30-KF-CM45 (2.0%) and Rhodobacter (0.9%) also contributed to ammonia removal at pH 10.0. On the cellular-level, bacteria enabled to maintain intracellular pH stabilization primarily through cation/proton antiporters, evidenced by significant increases in NhaA, TrkA and KefB activities by 98.0%, 151.7% and 115.2%, respectively. A 43.1% increase in carbonic anhydrase activity also facilitated consumption of aqueous OH- ions through biomineralization, leading to CaCO3 deposition on microbial surface. These findings further enhanced understandings of physiological adaptation of nitrifiers in the long-term alkaline activated sludge system.

Urban and agricultural land use led to niche differentiation of AOA, AOB and comammox along the Beiyun River continuum.

River ecological health has been severely threatened by anthropogenic land-use pressures. Here, by combining remote sensing and molecular biology methods, we evaluated the impact of land-use activities on nitrification, a fundamental ecological process in rivers, which is conducted by ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB), or the newly discovered complete ammonia oxidisers (comammox). We explored the relationships of the abundance, activity, and diversity of AOA, AOB, and comammox in river sediments with land-use pressure by proposing a quantitative land use pattern index (LPI) over a 184 km continuum along the Beiyun River in North China. We found that comammox dominated nitrification in the forestry upstream (67.07 % in summer, 56.40 % in winter), while AOB became the major player in the urban middle (56.51 % in summer, 53.08 % in winter) and agricultural downstream reaches (62.98 % in summer, 50.74 % in winter). In addition, urban and agricultural land use lowered the α diversity of AOA and comammox, as well as simplified their co-occurrence networks, but promoted AOB diversity and complicated their networks. The structural equation model illustrated that the key drivers affecting the key taxa and activities were ammonia, and C/N for AOB, and total organic matter, and pH for comammox. We thus conclude that watershed urban and agricultural land use drive the niche differentiation of AOA, AOB, and comammox, specifically leading to a robust AOB community but weakened AOA and comammox communities. Our study connects the macro and micro worlds and provides a new paradigm for studying the variation in microbial communities as well as the potential ecological consequences under the increased anthropogenic land-use pressures in the Anthropocene.

Nitrogen source influences the interactions of comammox bacteria with aerobic nitrifiers.

While the co-existence of comammox Nitrospira with canonical nitrifiers is well documented in diverse ecosystems, there is still a dearth of knowledge about the mechanisms underpinning their interactions. Understanding these interaction mechanisms is important as they may play a critical role in governing nitrogen biotransformation in natural and engineered ecosystems. In this study, we tested the ability of two environmentally relevant factors (nitrogen source and availability) to shape interactions between strict ammonia and nitrite-oxidizing bacteria and comammox Nitrospira in continuous flow column reactors. The composition of inorganic nitrogen species in reactors fed either ammonia or urea was similar during the lowest input nitrogen concentration (1 mg-N/L), but higher concentrations (2 and 4 mg-N/L) promoted significant differences in nitrogen species composition and nitrifier abundances. The abundance and diversity of comammox Nitrospira were dependent on both nitrogen source and input concentrations as multiple comammox Nitrospira populations were preferentially enriched in the urea-fed system. In contrast, their abundance was reduced in response to higher nitrogen concentrations in the ammonia-fed system. The preferential enrichment of comammox Nitrospira in the urea-fed system could be associated with their ureolytic activity calibrated to their ammonia oxidation rates, thus minimizing ammonia accumulation, which may be partially inhibitory. However, an increased abundance of comammox Nitrospira was not associated with a reduced abundance of nitrite oxidizers in the urea-fed system while a negative correlation was found between them in the ammonia-fed system, the latter dynamic likely emerging from reduced availability of nitrite to strict nitrite oxidizers at low ammonia concentrations. IMPORTANCE: Nitrification is an essential biological process in drinking water and wastewater treatment systems for treating nitrogen pollution. The discovery of comammox Nitrospira and their detection alongside canonical nitrifiers in these engineered ecosystems have made it necessary to understand the environmental conditions that regulate their abundance and activity relative to other better-studied nitrifiers. This study aimed to evaluate two important factors that could potentially influence the behavior of nitrifying bacteria and, therefore, impact nitrification processes. Column reactors fed with either ammonia or urea were systematically monitored to capture changes in nitrogen biotransformation and the nitrifying community as a function of influent nitrogen concentration, nitrogen source, and reactor depth. Our findings show that with increased ammonia availability, comammox Nitrospira decreased in abundance while nitrite oxidizers abundance increased. Yet, in systems with increasing urea availability, comammox Nitrospira abundance and diversity increased without an associated reduction in the abundance of canonical nitrifiers.

The response of structure and nitrogen removal function of the biofilm on submerged macrophytes to high ammonium in constructed wetlands.

The ammonium exceedance discharge from sewage treatment plants has a great risk to the stable operation of subsequent constructed wetlands (CWs). The effects of high ammonium shocks on submerged macrophytes and epiphytic biofilms on the leaves of submerged macrophytes in CWs were rarely mentioned in previous studies. In this paper, the 16S rRNA sequencing method was used to investigate the variation of the microbial communities in biofilms on the leaves of Vallisneria natans plants while the growth characteristics of V. natans plants were measured at different initial ammonium concentrations. The results demonstrated that the total chlorophyll and soluble sugar synthesis of V. natans plants decreased by 51.45% and 57.16%, respectively, and malondialdehyde content increased threefold after 8 days if the initial NH4+-N concentration was more than 5 mg/L. Algal density, bacterial quantity, dissolved oxygen, and pH increased with high ammonium shocks. The average removal efficiencies of total nitrogen and NH4+-N reached 73.26% and 83.94%, respectively. The heat map and relative abundance analysis represented that the relative abundances of phyla Proteobacteria, Cyanobacteria, and Bacteroidetes increased. The numbers of autotrophic nitrifiers and heterotrophic nitrification aerobic denitrification (HNAD) bacteria expanded in biofilms. In particular, HNAD bacteria of Flavobacterium, Hydrogenophaga, Acidovorax, Acinetobacter, Pseudomonas, Aeromonas, and Azospira had higher abundances than autotrophic nitrifiers because there were organic matters secreted from declining leaves of V. natans plants. The analysis of the nitrogen metabolic pathway showed aerobic denitrification was the main nitrogen removal pathway. Thus, the nitrification and denitrification bacterial communities increased in epiphytic biofilms on submerged macrophytes in constructed wetlands while submerged macrophytes declined under ammonium shock loading.

Blue-light irradiation induced partial nitrification.

The role of ray radiation from the sunlight acting on organisms has long-term been investigated. However, how the light with different wavelengths affects nitrification and the involved nitrifiers are still elusive. Here, we found more than 60 % of differentially expressed genes (DEGs) in nitrifiers were observed under irradiation of blue light with wavelengths of 440-480 nm, which were 13.4 % and 20.3 % under red light and white light irradiation respectively. Blue light was more helpful to achieve partial nitrification rather than white light or red light, where ammonium oxidization by ammonia-oxidizing archaea (AOA) with the increased relative abundance from 8.6 % to 14.2 % played a vital role. This was further evidenced by the enhanced TCA cycle, reactive oxygen species (ROS) scavenge and DNA repair capacity in AOA under blue-light irradiation. In contrast, nitrite-oxidizing bacteria (NOB) was inhibited severely to achieve partial nitrification, and the newly discovered encoded blue light photoreceptor proteins made them more sensitive to blue light and hindered cell activity. Ammonia-oxidizing bacteria (AOB) expressed genes for DNA repair capacity under blue-light irradiation, which ensured their tiny impact by light irradiation. This study provided valuable insights into the photosensitivity mechanism of nitrifiers and shed light on the diverse regulatory by light with different radiation wavelengths in artificial systems, broadening our comprehension of the nitrogen cycle on earth.

Detrimental effects of Cd and temperature on rice and functions of microbial community in paddy soils.

Heavy metal (HM) contamination and high environmental temperature (HT) are caused by anthropogenic activities that negatively impact soil microbial communities and agricultural productivity. Although HM contaminations have deleterious effects on microbes and plants; there are hardly any reports on the combined effects of HM and HT. Here, we reported that HT coupled with cadmium (Cd) accumulation in soil and irrigated water could seriously affect crop growth and productivity, alternatively influencing the microbial community and nutrient cycles of paddy soils in rice fields. We analyzed different mechanisms of plants and microflora in the rhizospheric region, such as plant rhizospheric nitrification, endophytes colonization, nutrient uptake, and physiology of temperature-sensitive (IR64) and temperature-resistant Huanghuazhan (HZ) rice cultivars against different Cd levels (2, 5 and 10 mg kg-1) with rice plants grown under 25 °C and 40 °C temperatures. Consequently, an increment in Cd accumulation was observed with rising temperature leading to enhanced expression of OsNTRs. In contrast, a greater decline in the microbial community was detected in IR64 cultivar than HZ. Similarly, ammonium oxidation, root-IAA, shoot-ABA production, and 16S rRNA gene abundance in the rhizosphere and endosphere were significantly influenced by HT and Cd levels, resulting in a significant decrease in the colonization of endophytes and the surface area of roots, leading to a decreased N uptake from the soil. Overall, the outcomes of this study unveiled the novel effects of Cd, temperature, and their combined effect on rice growth and functions of the microbial community. These results provide effective strategies to overcome Cd-phytotoxicity on the health of endophytes and rhizospheric bacteria in Cd-contaminated soil by using temperature-tolerant rice cultivars.

Influence of COD in Toxic Industrial Wastewater from a Chemical Concern on Nitrification Efficiency.

COD is an arbitrary indicator of the content of organic and inorganic compounds in wastewater. The aim of this research was to determine the effect of COD of industrial wastewater on the nitrification process. This research covered wastewater from acrylonitrile and styrene-butadiene rubbers, emulsifiers, polyvinyl acetate, styrene, solvents (butyl acetate, ethyl acetate) and owipian® (self-extinguishing polystyrene intended for expansion) production. The volume of the analyzed wastewater reflected the active sludge load in the real biological treatment system. This research was carried out by the method of short-term tests. The nitrification process was inhibited to the greatest extent by wastewater from the production of acrylonitrile (approx. 51%) and styrene-butadiene (approx. 60%) rubbers. In these wastewaters, nitrification inhibition occurred due to the high COD load and the presence of inhibitors. Four-fold dilution of the samples resulted in a two-fold reduction in the inhibition of nitrification. On the other hand, in the wastewater from the production of emulsifiers and polyvinyl acetate, a two-fold reduction in COD (to the values of 226.4 mgO2·dm-3 and 329.8 mgO2·dm-3, respectively) resulted in a significant decrease in nitrification inhibition. Wastewater from the production of styrene, solvents (butyl acetate, ethyl acetate) and owipian® inhibited nitrification under the influence of strong inhibitors. Lowering the COD value of these wastewaters did not significantly reduce the inhibition of nitrification.

Upgrading integrated fixed-biofilm activated sludge (IFAS) system into separated two-sludge denitrifying phosphorus removal system: Nutrient removal and microbial structure.

An integrated fixed-biofilm activated sludge (IFAS) system was upgraded into a separated two-sludge denitrifying phosphorus removal system to treat domestic sewage with a low carbon/nitrogen ratio (C/N = 3.6). The system was operated in IFAS mode for 135 days and then converted to the separated two-sludge mode to perform denitrifying phosphorus removal for 110 days. With denitrification and phosphorus removal conducted using "single carbon source", the experimental results show that the total inorganic nitrogen (TIN) and phosphorus (P) removal efficiencies were improved by 13.4% and 36.5%, respectively. As the activated sludge unit and biofilm unit were separated, Dechloromonas (4.10%) and Candidatus Accumulibacter (1.56%) were enriched in activated sludge system, while the nitrifiers Nitrospira and Nitrosomonas occupied the biofilm with proportions of 19.6% and 5.68%, respectively. The separated two-sludge denitrifying phosphorus removal system was characterized by higher nutrient removal and functional bacteria enrichment.

Urea fertilization and grass species alter microbial nitrogen cycling capacity and activity in a C4 native grassland.

Soil microbial transformation of nitrogen (N) in nutrient-limited native C4 grasslands can be affected by N fertilization rate and C4 grass species. Here, we report in situ dynamics of the population size (gene copy abundances) and activity (transcript copy abundances) of five functional genes involved in soil N cycling (nifH, bacterial amoA, nirK, nirS, and nosZ) in a field experiment with two C4 grass species (switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii)) under three N fertilization rates (0, 67, and 202 kg N ha-1). Diazotroph (nifH) abundance and activity were not affected by N fertilization rate nor grass species. However, moderate and high N fertilization promoted population size and activity of ammonia oxidizing bacteria (AOB, quantified via amoA genes and transcripts) and nitrification potential. Moderate N fertilization increased abundances of nitrite-reducing bacterial genes (nirK and nirS) under switchgrass but decreased these genes under big bluestem. The activity of nitrous oxide reducing bacteria (nosZ transcripts) was also promoted by moderate N fertilization. In general, high N fertilization had a negative effect on N-cycling populations compared to moderate N addition. Compared to big bluestem, the soils planted with switchgrass had a greater population size of AOB and nitrite reducers. The significant interaction effects of sampling season, grass species, and N fertilization rate on N-cycling microbial community at genetic-level rather than transcriptional-level suggested the activity of N-cycling microbial communities may be driven by more complex environmental factors in native C4 grass systems, such as climatic and edaphic factors.

Machine Learning Prediction of Nitrification From Ammonia- and Nitrite-Oxidizer Community Structure.

Accurately modeling nitrification and understanding the role specific ammonia- or nitrite-oxidizing taxa play in it are of great interest and importance to microbial ecologists. In this study, we applied machine learning to 16S rRNA sequence and nitrification potential data from an experiment examining interactions between cropping systems and rhizosphere on microbial community assembly and nitrogen cycling processes. Given the high dimensionality of microbiome datasets, we only included nitrifers since only a few taxa are capable of ammonia and nitrite oxidation. We compared the performance of linear and nonlinear algorithms with and without qPCR measures of bacterial and archaea ammonia monooxygenase subunit A (amoA) gene abundance. Our feature selection process facilitated the identification of taxons that are most predictive of nitrification and to compare habitats. We found that Nitrosomonas and Nitrospirae were more frequently identified as important predictors of nitrification in conventional systems, whereas Thaumarchaeota were more important predictors in diversified systems. Our results suggest that model performance was not substantively improved by incorporating additional time-consuming and expensive qPCR data on amoA gene abundance. We also identified several clades of nitrifiers important for nitrification in different cropping systems, though we were unable to detect system- or rhizosphere-specific patterns in OTU-level biomarkers for nitrification. Finally, our results highlight the inherent risk of combining data from disparate habitats with the goal of increasing sample size to avoid overfitting models. This study represents a step toward developing machine learning approaches for microbiome research to identify nitrifier ecotypes that may be important for distinguishing ecotypes with defining roles in different habitats.

Microbial communities contribute to the elimination of As, Fe, Mn, and NH4+ from groundwater in household sand filters.

Household sand filters (SFs) are widely applied to remove iron (Fe), manganese (Mn), arsenic (As), and ammonium (NH4+) from groundwater in the Red River delta, Vietnam. Processes in the filters probably include a combination of biotic and abiotic reactions. However, there is limited information on the microbial communities treating varied groundwater compositions and on whether biological oxidation of Fe(II), Mn(II), As(III), and NH4+ contributes to the overall performance of SFs. We therefore analyzed the removal efficiencies, as well as the microbial communities and their potential activities, of SFs fed by groundwater with varying compositions from low (3.3 µg L-1) to high (600 µg L-1) As concentrations. The results revealed that Fe(II)-, Mn(II)-, NH4+-, and NO2--oxidizing microorganisms were prevalent and contributed to the performance of SFs. Additionally, groundwater composition was responsible for the differences among the present microbial communities. We found i) microaerophilic Fe(II) oxidation by Sideroxydans in all SFs, with the highest abundance in SFs fed by low-As and high-Fe groundwater, ii) Hyphomicropbiaceae as the main Mn(II)-oxidizers in all SFs, iii) As sequestration on formed Fe and Mn (oxyhydr)oxide minerals, iv) nitrification by ammonium-oxidizing archaea (AOA) followed by nitrite-oxidizing bacteria (NOB), and v) unexpectedly, the presence of a substantial amount of methane monooxygenase genes (pmoA), suggesting microbial methane oxidation taking place in SFs. Overall, our study revealed diverse microbial communities in SFs used for purifying arsenic-contaminated groundwater, and our data indicate an important contribution of microbial activities to the key functional processes in SFs.

[Responses of soil ammonia-oxidizing microorganisms to simulated nitrogen deposition in a natural Castanopsis carlesii forest]. / äºšçƒ­å¸¦ç±³æ§ å¤©ç„¶æž—åœŸå£¤æ°¨æ°§åŒ–å¾®ç”Ÿç‰©å¯¹æ¨¡æ‹Ÿæ°®æ²‰é™çš„å“åº”.

Subtropical region of China is one of the global hotspots receiving nitrogen deposition. Nitrogen deposition could affect the abundance and community structure of ammonia oxidizers including ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA) and complete ammonia oxidizer (comammox Nitrospira), with consequences on soil nutrient cycling that are driven by microorganisms. There is limited understanding for the newly discovered comammox Nitrospira in the subtropical forest soils. Here, we investigated the effect of simulated N deposition on abundances of soil ammonia oxidizers in the Castanopsis fargesii Nature Reserve in Xinkou Town, Sanming City, Fujian Province, China. Soil samples were collected from the field plots which received long-term nitrogen deposition with different dosages, including: CK, no additional treatment; LN, low nitrogen deposition treatment, dosage of 40 kg N·hm-2·a-1; and HN, high nitrogen deposition treatment, dosage of 80 kg N·hm-2·a-1. After 8-year treatment, simulated N deposition decreased soil pH and organic matter content, and increased nitrate content. We failed to amplify the amoA gene of AOB in the tested soils. High nitrogen deposition increased the abundance of AOA, but did not affect the abundance of comammox Nitrospira clade A and clade B. The ratio of comammox Nitrospira to AOA decreased with N addition, indicating that N addition weakened the role of comammox Nitrospira in nitrification in the subtropical forest soils. However, there were strong non-specific amplifications for both comammox Nitrospira clades A and B, highlighting the demand for the development of high coverage and specificity primers for comammox Nitrospira investigations in the future. The abundance of comammox Nitrospira clade A was positively correlated with total nitrogen (TN) and NH4+ concentration, while that of clade B was positively associated with soil organic carbon (SOC), TN and NH4+ Concentration. Overall, our findings demonstrated that simulated N deposition increased the relative importance of AOA in nitrification in the natural Castanopsis carlesii forest soil. These findings could provide theoretical support in coping with global change and N deposition in these regions.

Biochemistry shapes growth kinetics of nitrifiers and defines their activity under specific environmental conditions.

Is it possible to find trends between the parameters that define microbial growth to help us explain the vast microbial diversity? Through an extensive database of kinetic parameters of nitrifiers, we analyzed if the dominance of specific populations of nitrifiers could be predicted and explained. We concluded that, in general, higher growth yield (YXS ) and ammonia affinity (a0NH3 ) and lower growth rate (µmax ) are observed for ammonia-oxidizing archaea (AOA) than bacteria (AOB), which would explain their considered dominance in oligotrophic environments. However, comammox (CMX), with the maximum energy harvest per mole of ammonia, and some AOB, have higher a0NH3 and lower µmax than some AOA. Although we were able to correlate the presence of specific terminal oxidases with observed oxygen affinities (a0O2 ) for nitrite-oxidizing bacteria (NOB), that correlation was not observed for AOB. Moreover, the presumed dominance of AOB over NOB in O2 -limiting environments is discussed. Additionally, lower statistical variance of a0O2 values than for ammonia and nitrite affinities was observed, suggesting nitrogen limitation as a stronger selective pressure. Overall, specific growth strategies within nitrifying groups were not identified through the reported kinetic parameters, which might suggest that mostly, fundamental differences in biochemistry are responsible for underlying kinetic parameters.

Dimethylpyrazole-based nitrification inhibitors have a dual role in N2O emissions mitigation in forage systems under Atlantic climate conditions.

Nitrogen fertilization is the most important factor increasing nitrous oxide (N2O) emissions from agriculture, which is a powerful greenhouse gas. These emissions are mainly produced by the soil microbial processes of nitrification and denitrification, and the application of nitrification inhibitors (NIs) together with an ammonium-based fertilizer has been proved as an efficient way to decrease them. In this work the NIs dimethylpyrazole phosphate (DMPP) and dimethylpyrazole succinic acid (DMPSA) were evaluated in a temperate grassland under environmental changing field conditions in terms of their efficiency reducing N2O emissions and their effect on the amount of nitrifying and denitrifying bacterial populations responsible of these emissions. The stimulation of nitrifying bacteria induced by the application of ammonium sulphate as fertilizer was efficiently avoided by the application of both DMPP and DMPSA whatever the soil water content. The denitrifying bacteria population capable of reducing N2O up to N2 was also enhanced by both NIs provided that sufficiently high soil water conditions and low nitrate content were occurring. Therefore, both NIs showed the capacity to promote the denitrification process up to N2 as a mechanism to mitigate N2O emissions. DMPSA proved to be a promising NI, since it showed a more significant effect than DMPP in decreasing N2O emissions and increasing ryegrass yield.

Nitrifiers drive successions of particulate organic matter and microbial community composition in a starved macrocosm.

Organic carbon produced by nitrifiers plays an important role in maintaining the microbial metabolism in the aphotic ocean layer with carbon and energy scarcity. However, the contribution of nitrifiers to organic carbon processing remains unclear. To explore how nitrification impacts the material cycle in the starved ecosystem, we set up an ultra-large volume, long-term incubation experiment. Seawater collected from the Halifax coastal ocean was pumped into the Aquatron Tower Tank located at Dalhousie University, Canada, and was incubated under dark conditions for 73 days. The results indicated that the relative abundance of nitrifiers increased and nitrification was strengthened in the dark system where energy and organic carbon were scarce. The importance of nitrogenous compounds in particulate materials increased over the course of the incubation. Correlation analysis showed that the relative abundances of nitrifiers and particulate organic compounds containing nitrogen were significantly and positively correlated. Furthermore, network analysis suggested that metabolic processes related to nitrogenous and aromatic compounds are most important to particle associated bacteria. This study suggests that the nitrifiers could produce a series of organic compounds that result in the alteration of organic matter composition by promoting the degradation of recalcitrant aromatic compounds, which has important implications for organic matter processing in the starved dark ecosystem.

Impact of irrigation with fish-processing effluents on nitrification and ammonia-oxidizer abundances in Patagonian arid soils.

This study aimed to evaluate the short-term effects of irrigation with diluted fish-processing effluents on soil pH, electrical conductivity, nitrification rate and abundance of ammonia oxidizers. To accomplish that, we constructed microcosms of soil from an undisturbed arid ecosystem of Patagonia, and irrigated them for 2 months with diluted effluents from a fish-processing factory or with water as control. In the initial soil sample, and along the experiment, we determined soil pH, electrical conductivity, and the concentration of inorganic nitrogen forms, which we used to calculate the net nitrification rate. We further estimated the abundances of ammonia-oxidizing archaea and bacteria in the initial soil sample and at the end of the experiment, by qPCR of amoA genes. Soil pH decreased and electrical conductivity increased in both irrigation treatments, although the effect was higher in effluent-irrigated microcosms. Soil nitrate + nitrite concentration, and thus the nitrification rate, was higher in effluent than in water-irrigated microcosms. The abundance of archaeal amoA genes was higher under effluent than water-irrigation, but that of bacterial amoA genes did not vary significantly between treatments. Neither ammonia-oxidizing archaea nor bacteria were influenced by the changes in soil pH and electrical conductivity induced by effluent irrigation.

In-situ expressions of comammox Nitrospira along the Yangtze River.

The recent discovery of comammox Nitrospira as complete nitrifiers has significantly enriched our understanding on the nitrogen cycle, yet little is known about their metabolic transcripts in natural aquatic ecosystems. Using the genome-centric metatranscriptomics, we provided the first in-situ expression patterns of comammox Nitrospira along the Yangtze River. Our study confirmed widespread expressions of comammox Nitrospira, with the highest transcription accounting for 33.3% and 63.8% of amoA and nxrAB genes expressed in ammonia-oxidizing prokaryotes (AOPs) and Nitrospira sublineages I/II, respectively. Moreover, comammox two clades differed in nitrification, with clade A acting as the dominator to ammonia oxidation in comammox, and clade B contributing more transcripts to nitrite oxidation than to ammonia oxidation. Compared to canonical Nitrospira, comammox community had lower expressions of ammonia/nitrite transporters and nitrogen assimilatory genes, but far higher expressions in urea transport and hydrolysis, facilitating to derivation of ammonia and energy mainly through intracellular ureolytic metabolism. This suggests no need for "reciprocal-feeding" between canonical Nitrospira and AOPs in a natural river. Aerobic mixotrophy of comammox bacteria was suggested by expressions of genes coding for respiratory complexes I-V, oxidative/reductive TCA cycle, oxygen stress defenses, and transport/catabolism of simple carbohydrates and low-biosynthetic-cost amino acids. Intriguingly, significant positive correlations among expressions of ammonia monooxygenases, hydroxylamine dehydrogenase and copper-dependent nitrite reductase indicated that comammox Nitrospira had the potential of converting nitrite to nitric oxide accompanied by ammonia oxidation under low-C/N and aerobic conditions, while gene expressions in this pathway were significantly and positively associated with pH. Overall, this study illustrated novel transcriptional characteristics of comammox Nitrospira, and highlighted the necessity of reassessing their contributions to biogeochemical carbon and nitrogen cycling with perspective of in-situ meta-omics as well as culture experiments.

Application of glucose for improving NH4+-N removal in micro-polluted source water by immobilized heterotrophic nitrifiers at low temperature.

Bio-enhanced activated carbon (BEAC) filters have shown potential in source water purification. The key drawback of this system is the difficulty of the set-up at low temperature. Here, glucose was applied to help immobilize more functional heterotrophic nitrifiers and further improve NH4+-N removal by BEAC. Results showed that pre-loading glucose on granular activated carbon could achieve better immobilization efficiency with 5.12 × 108 CFU/g-DW C biomass and 3.77 mg TF/L/g-DW C dehydrogenase activity after artificial immobilization, which were separately 12.5 and 4.2 times of the control. 95-d running data at different conditions showed the superiority of both immobilization and NH4+-N removal could last and defend environment changes during relatively long period. Even at the end of operating, the abundance of targeting genus (Acinetobacter) still occupied 9.59% of microbial communities on BEAC, while this value was only 1.24% without pre-loading glucose. Biolog-ECO plate analysis found pre-loading glucose improved organic nitrogen metabolism effectively, along with carbohydrate, amino, alcohol, amine and carboxylic acid metabolism on BEAC.

[Seasonal Effects of Influent Ammonia Oxidizing Bacteria of Municipal Wastewater Treatment Plants on Activated Sludge System].

The specific ammonia uptake rates (SAUR) and ammonia oxidizing bacteria (AOB) community of influent sewage and activated sludge in the 2nd wastewater treatment plant (WWTP) of Xi'an without the primary settling tank were analyzed over multiple years to explore the seasonal effects of the influent AOB on the activated sludge systems. During the experiment, the SAUR of the raw sewage and activated sludge were 0.48-3.02 mg·(g·h)-1 and 0.68-2.25 mg·(g·h)-1, respectively. Meanwhile, the correlation analysis indicated that the monthly SAUR of the raw sewage was highly correlated with that of the activated sludge of the following month (r=0.862,P<0.05), which indicated that influent nitrifiers had a significant effect on the nitrification performance of activated sludge. Considering that the estimated AOB seeding intensities based on the ammonia oxidizing activity were 0.21-0.92 g·(g·d)-1, the nitrifier immigration from the raw sewage should added to the design of WWTP and the activated sludge modeling. Moreover, the qPCR results revealed that the AOB abundance of activated sludge in winter decreased but remained at 1010 cells·g-1, indicating that the immigration of influent nitrifiers could partially compensate for the reduction of the AOB abundance in the activated sludge caused by decreasing temperatures. Finally, the Illumina MiSeq sequencing demonstrated that the shared dominant AOB between the raw sewage and activated sludge were Nitrosomonas sp. Nm58, Nitrosomonas sp. JL21, and bacterium CYCU-0253. These findings can provide theoretical support for the design and operation of a WWTP.

The contrasting effects of biochar and straw on N2O emissions in the maize season in intensively farmed soil.

This study evaluated the combined effects of biochar and straw on N2O flux and the community compositions of nitrifiers and denitrifiers in the maize season in an intensively farmed area in northern China. The experiment consisted of four treatments: (1) CK (only chemical fertilizer application); (2) C (biochar application); (3) SR (straw application to the field); and (4) C+SR (the application of both biochar and straw). The results indicated that during the maize growing season, N2O flux decreased by 30.3% in the C treatment and increased by 13.2% and 37.0% in the SR and C+SR treatments compared with CK, respectively. NO3--N, NH4+-N, and microbial biomass carbon (MBC) were the main soil factors affecting N2O flux, and they were positively correlated with NO3--N and negatively correlated with MBC in the C treatment and positively correlated with NH4+-N in the SR and C+SR treatments. Both biochar addition and straw return shifted the community compositions of nitrifiers and denitrifiers. N2O production was mainly reduced by promoting the ammonia-oxidizing bacteria (AOB) gene abundance and inhibiting the nirK gene abundance in the C treatment but promoted by inhibiting the AOB and nosZ gene abundances in the SR and C+SR treatments. Nitrosospira (AOB) and Rhizobium (nirK) were the main contributors among the treatments. NO3--N, NH4+-N, and MBC were the main soil factors affecting the denitrifier communities. The predominant species associated with the nirK, nirS, and nosZ genes were positively correlated with NO3--N and MBC and negatively correlated with NH4+-N. These results provide valuable information on the mechanism of N2O production and reduction in biochar- and straw-amended soil under field conditions.