Nitrification Denitrification Research Articles

Nanoparticles of Zinc Oxides Mitigated N2O Emissions in Tea Plantation Soil

The excessive application of nitrogen in tea plantations leads to severe soil acidification and N2O emission boosting. To promote sustainable agriculture, nanoparticles (NPs) have emerged as alternative fertilizers, but their effects on soil nitrification and greenhouse gas emissions in tea plantations remain unclear. In this study, the effects of NP type (ZnO-NPs and Fe2O3-NPs) and dose (0, 1, 10, and 100 mg·kg⁻1) on soil N2O emissions were investigated via a lab incubation trial. Soil pH, ammonium, and nitrate changes were also monitored during the incubation period. The abundance of functional genes related to nitrification and denitrification processes was analyzed as well. The results showed that ZnO-NPs led to a decrease in N2O emissions. The reduction effect was stronger with increasing dose and resulted in a 33% reduction at an addition rate of 100 mg·kg−1. The cumulative N2O emissions had significantly positive correlations with NH4+-N and NO3−-N. ZnO-NP addition showed a significantly negative effect on Ammonia-Oxidizing Archaea (AOA) but a positive effect on Ammonia-Oxidizing Bacteria (AOB) gene abundance. In contrast, Fe2O3-NPs showed an insignificant impact on N2O emissions and soil N content, as well as nitrification–denitrification gene abundance, regardless of different doses. These results imply that the application of ZnO-NPs may inhibit nitrification through the retarding of AOA activity. This study provided us with a potential practice to reduce N2O emissions in tea plantations by applying ZnO-NPs, but the efficiency of this reduction needs further examination under ambient conditions before field application.

Nitrification–Autotrophic Denitrification Using Elemental Sulfur as an Electron Donor in a Sequencing Batch Reactor (SBR): Performance and Kinetic Analysis

Simultaneous nitrification and autotrophic denitrification (SNAD) has received attention as an efficient biological nitrogen removal alternative. However, SNAD using elemental sulfur (S0) has scarcely been studied. Thus, the main objective of this research was to study the behavior of a simultaneous nitrification–autotrophic denitrification operation in a sequential batch reactor (SNAD-SBR) at a lab scale using S0 as an electron donor, including its kinetics. Two-scale reactors were operated at lab scales in cycles for 155 days with an increasing nitrogen loading rate (NLR: 0.0296 to 0.0511 kg N-NH4+/m3/d) at 31 °C. As a result, simultaneous nitrification–autotrophic denitrification using S0 as an electron donor was performed successfully, with nitrification efficiency of 98.63% and denitrification efficiency of 44.9%, with autotrophic denitrification as the limiting phase. The kinetic model adjusted for ammonium-oxidizing bacteria (AOB) was the Monod-type kinetic model (µmax = 0.791 d−1), while, for nitrite-oxidizing bacteria (NOB), the Haldane-type model was employed (µmax = 0.822 d−1). For denitrifying microorganisms, the kinetic model was adjusted by a half order (k1/2v = 0.2054 mg1/2/L1/2/h). Thus, we concluded that SNAD could be feasible using S0 as an electron donor, with kinetic behavior similar to that of other processes.

Genetic variation in Zea mays influences microbial nitrification and denitrification in conventional agroecosystems

Abstract Background and Aims Nitrogenous fertilizers provide a short-lived benefit to crops in agroecosystems, but stimulate nitrification and denitrification, processes that result in nitrate pollution, N2O production, and reduced soil fertility. Recent advances in plant microbiome science suggest that genetic variation in plants can modulate the composition and activity of rhizosphere N-cycling microorganisms. Here we attempted to determine whether genetic variation exists in Zea mays for the ability to influence the rhizosphere nitrifier and denitrifier microbiome under “real-world” conventional agricultural conditions. Methods To capture an extensive amount of genetic diversity within maize we grew and sampled the rhizosphere microbiome of a diversity panel of germplasm that included ex-PVP inbreds (Z. mays ssp. mays), ex-PVP hybrids (Z. mays ssp. mays), and teosinte (Z. mays ssp. mexicana and Z. mays ssp. parviglumis). From these samples, we characterized the microbiome, a suite of microbial genes involved in nitrification and denitrification and carried out N-cycling potential assays. Results Here we are showing that populations/genotypes of a single species can vary in their ecological interaction with denitrifers and nitrifers. Some hybrid and teosinte genotypes supported microbial communities with lower potential nitrification and potential denitrification activity in the rhizosphere, while inbred genotypes stimulated/did not inhibit these N-cycling activities. These potential differences translated to functional differences in N2O fluxes, with teosinte plots producing less GHG than maize plots. Conclusion Taken together, these results suggest that Zea genetic variation can lead to changes in N-cycling processes that result in N leaching and N2O production, and thereby are selectable targets for crop improvement. Understanding the underlying genetic variation contributing to belowground microbiome N-cycling into our conventional agricultural system could be useful for sustainability.

Treatment of Pickle Wastewater under Varying Salinity Conditions within the Sequencing Batch Biofilm Reactor System

Treatment of Pickle Wastewater under Varying Salinity Conditions within the Sequencing Batch Biofilm Reactor System

Conversion behavior of heterotrophic nitrification - aerobic denitrification bacterium Paracoccus denitrificans HY-1 in nitrogen and phosphorus removal

The heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria harbored remarkable removal capacity in eliminating single nitrogen pollutant, but their comprehensive performance in dealing with mixed nitrogen sources remained unknown. In this study, the denitrification capacity of HN-AD bacteria Paracoccus denitrificans HY-1 was explored through whole-genome sequencing and nitrogen metabolic pathway analysis, and the complex nitrogen sources with gradient concentration were used to validate its nitrogen removal efficiency under both aerobic and anaerobic conditions. The results demonstrated that strain HY-1 harbored an efficient degradation capacity on three nitrogen sources, and the removal rates of NH4+-N, NO2−-N and NO3−-N were 10.67 mg·L−1·h−1, 27.81 mg·L−1·h−1, and 22.05 mg·L−1·h−1, respectively. The denitrification rate of 15.33 mg·L−1·h−1 was obtained under anaerobic condition with mixed nitrogen sources. Also, the simultaneous nitrification denitrification phosphorus removal (SNDPR) trial revealed that strain HY-1 could eliminate PO43−-P with a rate of 0.38 mg·L−1·h−1 under coexistence condition of the three nitrogen sources. Combined with the nitrogen balance analysis, the results indicated that strain HY-1 possesses a great potential for application in complex nitrogen and phosphorus contaminated environments.

Harnessing the Potential of Extracellular Polymeric Substances in Enhancing ANAMMOX Processes: Mechanisms, Strategies, and Perspectives

Anaerobic ammonium oxidation (ANAMMOX) has emerged as a promising sustainable nitrogen removal technology that offers significant advantages over conventional nitrification–denitrification processes, such as reduced energy consumption, a 60% reduction in oxygen demand, and a 90% reduction in sludge production. However, the practical application of ANAMMOX is hindered by several challenges, including the slow growth of ANAMMOX bacteria, long start-up periods, and high sensitivity to environmental disturbances. Recent studies have highlighted the crucial role of extracellular polymeric substances (EPSs) in the formation, activity, and stability of ANAMMOX biofilms and granules. An EPS is a complex mixture of high-molecular-weight polymers secreted by microorganisms, mainly composed of polysaccharides, proteins, nucleic acids, and lipids. The diverse physicochemical properties and functional groups of EPSs enable them to serve as a structural scaffold, protective barrier, sorption site, electron shuttle, and nutrient source for ANAMMOX bacteria. This review aims to provide an overview of the latest research progress on harnessing the potential of EPSs to enhance the ANAMMOX process. The characteristics, compositions, and extraction methods of ANAMMOX-derived EPSs are summarized. The mechanisms of how EPSs facilitate the enrichment, immobilization, aggregation, and adaptation of ANAMMOX bacteria are elucidated. The strategies and effects of EPS supplementation on improving the performance and robustness of ANAMMOX reactors under various stresses are critically reviewed. The challenges and future perspectives of the EPS-mediated optimization of the ANAMMOX process are also discussed. This review sheds new light on exploiting EPSs as a renewable bioresource to develop more efficient and stable ANAMMOX applications for sustainable wastewater treatment.

Simultaneous removal of ammonia nitrogen, calcium and cadmium in a biofilm reactor based on microbial-induced calcium precipitation: Optimization of conditions, mechanism and community biological response

With the enhancement of environmental governance regulations, the discharge requirements for reverse osmosis wastewater have become increasingly stringent. This study proposes an innovative approach utilizing heterotrophic nitrification and aerobic denitrification (HNAD)-based biomineralization technology, combined with coconut palm silk loaded biochar, to offer a novel solution for resource-efficient and eco-friendly treatment of reverse osmosis wastewater. Zobellella denitrificans sp. LX16 were loaded onto modified coir silk and showed removal efficiencies of up to 97.38, 94.58, 86.24, and 100% for NH4+-N (65 mg L−1), COD (900 mg L−1), Ca2+ (180 mg L−1), and Cd2+ (25 mg L−1). Analysis of the metabolites of microorganisms reveals that coconut palm silk loaded with deciduous biochar (BCPS) not only exerts a protective effect on microorganisms, but also enhances their growth, metabolism, and electron transfer capabilities. Characterization of precipitation phenomena elucidated the mechanism of Cd2+ removal via ion exchange, precipitation, and adsorption. Employing high-throughput and KEGG functional analyses has confirmed the biota environmental response strategies and the identification of key genes like HNAD.

Nitrogen removal and carbon reduction of mature landfill leachate under extremely low dissolved oxygen conditions by simultaneous partial nitrification anammox and denitrification

In this study, a SNAD-SBBR process was implemented to achieve ammonia removal and carbon reduction of mature landfill leachate under extremely low dissolved oxygen conditions (0.051 mg/L) for a continuous operation of 266 days. The process demonstrated excellent removal performance, with ammonia nitrogen removal efficiency reaching 100 %, total nitrogen removal efficiency reaching 87.56 %, and an average removal rate of 0.180 kg/(m3·d). The recalcitrant organic compound removal efficiency reached 34.96 %. Nitrogen mass balance analysis revealed that the Anammox process contributed to approximately 98.1 % of the nitrogen removal. Candidatus Kuenenia achieved a relative abundance of 1.49 % in the inner layer of the carrier. In the SNAD-SBBR system, the extremely low DO environment created by the highly efficient partial nitrification stage enabled the coexistence of AnAOB, denitrifying bacteria, and Nitrosomonas, synergistically achieving ammonia removal and carbon reduction. Overall, the SNAD-SBBR process exhibits low-cost and high-efficiency characteristics, holding tremendous potential for landfill leachate treatment.

Nitrate denitrification rate response to temperature gradient change during river bank infiltration.

Nitrate attenuation during river bank infiltration is the key process for reducing nitrogen pollution. Temperature is considered to be an important factor affecting nitrate attenuation. However, the magnitude and mechanism of its impact have not been clear for a long time. In this study, the effects of temperature and temperature gradient on the nitrate denitrification rate were investigated via static batch and dynamic soil column simulation experiments. The results showed that temperature had a significant effect on the denitrification rate. Temperature effects were first observed in denitrifying bacteria. At low temperatures, the microorganism diversity was low, resulting in a lower denitrification rate constant. The static experimental results showed that the denitrification rate at 19°C was approximately 2.4 times that at 10°C. The dynamic soil column experiment established an exponential positive correlation between the nitrate denitrification decay kinetic constant and temperature. The affinity of denitrifying enzymes for nitrate in the reaction substrate was ordered as follows: decreasing temperature gradient (30°C → 10°C) > zero temperature gradient (10°C) > increasing temperature gradient condition (0°C → 10°C). This study provides a theoretical basis for the biogeochemical processes underlying river bank infiltration, which will help aid in the development and utilization of groundwater resources.

Research Progress,Problems and Future Prospects of A New Combined Anaerobic Ammonia Oxidation and Nitrogen Removal Process

Anaerobic ammonium oxidation (Anammox) is a cost-effective and efficient process for nitrogen removal, which has attracted widespread attention in the field of wastewater treatment due to its significant advantages. However, the slow rate of growth of anaerobic ammonium oxidizing bacteria (AnAOB) and its sensitivity to operational and environmental conditions hinder its wide application. Although various control strategies have been explored and developed, the application of Anammox process still faces many challenges, including unstable nitrogen removal performance and the necessity for additional treatment of effluent nitrate nitrogen. Due to the importance and necessity of Anammox in wastewater denitrification, this paper first introduces the present biological denitrification system based on Anammo. In view of the insufficient source of nitrite nitrogen in nitrogen-containing wastewater, nitrite nitrogen can be provided by partial nitrification (PN) or partial denitrification (PD) , however, the stability of system operation and efficiency of denitrification are still the difficulties in Anammox engineering applications. It is especially crucial to study the combined new processes based on Anammox to achieve stable deep biological nitrogen removal from wastewater. Therefore, this paper focuses on latest research progress in Anammox coupled simultaneous partial nitrification, denitrification and anammox (SPNDA) , constructed wetland (CW) , phosphorus removal/ recovery process, sulfur denitrification removal (SDA) and denitrifying anaerobic methane oxidation (ADAMO) . The nitrogen removal performance, influencing factors, operation advantages, research progress, application prospects and challenges are reviewed. Compared with the single Anammox process, the combined process offers several advantages, including overcoming the issue of nitrate in the effluent (SPNDA, A-DAMO, SDA) , alleviate the inhibition of organic matter on AnAOB (SPNDA, SDA) , recover non-renewable resources (HAP) , and achieve a clean environment with significant economic benefits (A-CW) , providing flexibility for the promotion and application of Anammox process. The study outlines future directionsfor researching, developing, and applying the Anammox combined nitrogen removal process, and at the same time provides as a reference for lowering energy consumption, achieving efficient and stable nitrogen removal processes, and broadensing the scope of engineering application.

Start-up of pilot-scale ANAMMOX reactor for low-carbon nitrogen removal from anaerobic digestion effluent of kitchen waste

The pilot-scale simultaneous denitrification and methanation (SDM)-partial nitrification (PN)-anaerobic ammonia oxidation (Anammox) system was designed to treat anaerobic digestion effluent of kitchen waste (ADE-KW). The SDM-PN was first started to avoid the inhibition of high-concentration pollutants. Subsequently, Anammox was coupled to realize autotrophic nitrogen removal. Shortcut nitrification–denitrification achieved by the SDM-PN. The NO2–-N accumulation (92 %) and NH4+-N conversion (60 %) were achieved by PN, and the removal of TN and COD from the SDM-PN was 70 % and 73 %, respectively. After coupling Anammox, the TN (95 %) was removed with a TN removal rate of 0.51 kg·m−3·d-1. Microbiological analyses showed a shift from dominance by Methanothermobacter to co-dominance by Methanothermobacter, Thermomonas, and Flavobacterium in SDM during the SDM-PN. While after coupling Anammox, Candidatus kuenenia was enriched in the Anammox zone, the SDM zone shifted back to being dominated by Methanothermobacter. Overall, this study provides new ideas for the treatment of ADE-KW.

Enhancing swine manure treatment: A full-scale techno-economic assessment of nitrogen recovery, pure oxygen aeration and effluent polishing

In regions with intensive livestock production, managing the environmental impact of manure is a critical challenge. This study, set in Flanders (Belgium), evaluates the effectiveness of integrating process intensification measures into the treatment of piggery manure to mitigate nitrogen (N) surplus issues. The research investigates the techno-economic benefits of implementing three key interventions: pure oxygen (PO) aeration, ammonia (NH3) stripping-scrubbing (SS) pretreatment, and tertiary treatment using constructed wetlands (CW), within the conventional nitrification-denitrification (NDN) process.Conducted at a full-scale pig manure treatment facility, our analysis employs steady-state mass balances for N and phosphorus (P) to assess the impact of these process intensification strategies. Findings indicate that the incorporation of advanced treatment steps significantly enhances the efficiency and cost-effectiveness of the manure management system. Specifically, the application of PO aeration is shown to reduce overall treatment costs by nearly 4%, while the addition of an NH3 SS unit further decreases expenses by 1–2%, depending on the counter acid utilized. Moreover, the implementation of a CW contributes an additional 4% in cost savings.Collectively, these measures offer substantial improvements in processing capacity, reduction of by-product disposal costs, and generation of additional revenue from high-quality fertilising products. The study highlights the potential of advanced treatment technologies to provide economically viable and environmentally sustainable solutions for manure management in livestock-dense regions, emphasizing the cumulative economic benefit of a holistic approach to process intensification (10%).

Promoting denitrification via high electron conversion ratio in the iron valence Cycle: Prospects for Pseudomonas sp. JM-7 application in environmental engineering

The interdependence between iron and nitrogen in the environment is of utmost importance, and this study has explored the intricate relationship between denitrification and iron valence cycling using Pseudomonas sp. strain JM-7 (P. JM-7) as a model bacteria. Our research demonstrated that P. JM-7 had the ability to effectuate electron transfer from the periplasmic space to electron acceptors, such as nitrate, via Cytochrome c-proteins (c-Cyts). We found that iron valence state transformations with Fe(II)/Fe(III) promoted P. JM-7′s denitrification of nitrate coupled to yeast extract powder (YEP) oxidation. Denitrification rates depended on temperature (>21 °C) and pH (6–9). During logarithmic growth, 5 g/L YEP enabled 350 mg/L nitrate removal, increasing to 1650 mg/L in stable growth. Addition of 4 mM Fe(II)/Fe(III) increased denitrification to 500 mg/L during logarithmic growth. The dynamic test results displayed that compared to the SiO2 filler control group, iron-containing fillers could facilitate P. JM-7 to reduce 100 mg/L nitrate with 50 mg/L YEP electrons, corresponding to 6 times the maximum electron transfer efficiency in the static test. Overall, iron redox cycling significantly enhanced P. JM-7 denitrification, imparting insights into iron–nitrogen interactions. In summary, our research imparts valuable insights into the nexus between iron and nitrogen in the environment, highlighting the potential application of P. JM-7 in environmental engineering. Our findings indicate that P. JM-7 exhibits a significant denitrification capability, and further research is recommended to investigate its ability to remove phosphorus and promote sustainable environmental remediation practices.

Retrofitting a poorly designed system into a full performing nitrogen‐removal activated sludge process—a scientific challenge

AbstractBACKGROUNDThis study intended to detect the inherent deficiencies of prescription‐type empirical design procedures often adopted for nitrogen ‐removal activated sludge systems. Principles of process stoichiometry and kinetics were used to identify a series of hotspots that were likely to impair and disrupt system performance and efficiency. Available data, derived from an existing plant subject to a similar design approach, were utilized to numerically assess significant hotspots and to define a sustainable remedial action in compliance with applicable scientific principles.RESULTSCritical appraisal of the implemented design exposed a large spectrum of deficiencies affecting system efficiency and performance, including nitrification efficiency, denitrification potential, anoxic volume and sludge age, internal recycling and nitrate removal. The major role of the inorganic suspended solids as a design parameter was also ignored. The remedial strategy adopted for retrofitting corrected all the observed discrepancies and it devised a sustainable approach of flipping around the existing reactors so that they could be operated as a series of carousel/fast recycle reactors, each including anoxic and aerobic volume fractions to provide full nitrification.CONCLUSIONThe merit of the proposed remedial strategy was justified by offering two additional options of equally effective system operation with either reduced reactor volume or increased sewage flow. The study presented conclusive evidence to show why prescription‐type design should be avoided, providing numerical proof that this methodology is incompatible with stoichiometric and kinetic characteristics of sewage under local conditions. © 2024 Society of Chemical Industry (SCI).

Research progress of anaerobic ammonium oxidation (Anammox) process based on integrated fixed-film activated sludge (IFAS).

The integrated fixed-film activated sludge (IFAS) process is considered one of the cutting-edge solutions to the traditional wastewater treatment challenges, allowing suspended sludge and attached biofilm to grow in the same system. In addition, the coupling of IFAS with anaerobic ammonium oxidation (Anammox) can further improve the efficiency of biological denitrification. This paper summarises the research progress of IFAS coupled with the anammox process, including partial nitrification anammox, simultaneous partial nitrification anammox and denitrification, and partial denitrification anammox technologies, and describes the factors that limit the development of related processes. The effects of dissolved oxygen, influent carbon source, sludge retention time, temperature, microbial community, and nitrite-oxidising bacteria inhibition methods on the anammox of IFAS are presented. At the same time, this paper gives an outlook on future research focus and engineering practice direction of the process.

Methylomarinovum tepidoasis sp. nov., a moderately thermophilic methanotroph of the family Methylothermaceae isolated from a deep-sea hydrothermal field.

A novel aerobic methanotrophic bacterium, designated as strain IN45T, was isolated from in situ colonisation systems deployed at the Iheya North deep-sea hydrothermal field in the mid-Okinawa Trough. IN45T was a moderately thermophilic obligate methanotroph that grew only on methane or methanol at temperatures between 25 and 56 °C (optimum 45-50 °C). It was an oval-shaped, Gram-reaction-negative, motile bacterium with a single polar flagellum and an intracytoplasmic membrane system. It required 1.5-4.0 % (w/v) NaCl (optimum 2-3 %) for growth. The major phospholipid fatty acids were C16 : 1ω7c, C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The 16S rRNA gene sequence comparison revealed 99.1 % sequence identity with Methylomarinovum caldicuralii IT-9T, the only species of the genus Methylomarinovum with a validly published name within the family Methylothermaceae. The complete genome sequence of IN45T consisted of a 2.42-Mbp chromosome (DNA G+C content, 64.1 mol%) and a 20.5-kbp plasmid. The genome encodes genes for particulate methane monooxygenase and two types of methanol dehydrogenase (mxaFI and xoxF). Genes involved in the ribulose monophosphate pathway for carbon assimilation are encoded, but the transaldolase gene was not found. The genome indicated that IN45T performs partial denitrification of nitrate to N2O, and its occurrence was indirectly confirmed by N2O production in cultures grown with nitrate. Genomic relatedness indices between the complete genome sequences of IN45T and M. caldicuralii IT-9T, such as digital DNA-DNA hybridisation (51.2 %), average nucleotide identity (92.94 %) and average amino acid identity (93.21 %), indicated that these two methanotrophs should be separated at the species level. On the basis of these results, strain IN45T represents a novel species, for which we propose the name Methylomarinovum tepidoasis sp. nov. with IN45T (=JCM 35101T =DSM 113422T) as the type strain.

Transcriptomic analysis of nitrogen metabolism pathways in Klebsiella aerogenes under nitrogen-rich conditions.

The acceleration of the nitrogen cycle and the nitrogen excess observed in some coastal waters has increased interest into understanding the biochemical and molecular basis of nitrogen metabolism in various microorganisms. To investigate nitrogen metabolism of a novel heterotrophic nitrification and aerobic denitrification bacterium Klebsiella aerogenes strain (B23) under nitrogen-rich conditions, we conducted physiological and transcriptomic high-throughput sequencing analyses on strain B23 cultured on potassium nitrate-free or potassium nitrate-rich media. Overall, K. aerogenes B23 assimilated 82.47% of the nitrate present into cellular nitrogen. Further, 1,195 differentially expressed genes were observed between K. aerogenes B23 cultured on potassium nitrate-free media and those cultured on potassium nitrate-rich media. Gene annotation and metabolic pathway analysis of the transcriptome were performed using a series of bioinformatics tools, including Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Non-Redundant Protein Database annotation. Accordingly, the nitrogen metabolism pathway of K. aerogenes B23 was analyzed; overall, 39 genes were determined to be involved in this pathway. Differential expression analysis of the genes involved in the nitrogen metabolism pathway demonstrated that, compared to the control, FNR, NarK/14945, fdx, gshA, proB, proA, gapA, argH, artQ, artJ, artM, ArgR, GAT1, prmB, pyrG, glnS, and Ca1 were significantly upregulated in the nitrogen-treated K. aerogenes B23; these genes have been established to be involved in the regulation of nitrate, arginine, glutamate, and ammonia assimilation. Further, norV, norR, and narI were also upregulated in nitrogen-treated K. aerogenes B23; these genes are involved in the regulation of NO metabolism. These differential expression results are important for understanding the regulation process of key nitrogen metabolism enzyme genes in K. aerogenes B23. Therefore, this study establishes a solid foundation for further research into the expression regulation patterns of nitrogen metabolism-associated genes in K. aerogenes B23 under nitrogen-rich conditions; moreover, this research provides essential insight into how K. aerogenes B23 utilizes nutritional elements.

In situ membrane separation drives nitrous oxide enrichment from nitrate denitrification for energy recovery

Efficiently enriching nitrous oxide (N2O) can provide a foundation for energy recovery during wastewater treatment. In this study, gas-liquid separation membranes were placed in a sequencing batch reactor fed with acidic nitrate-containing wastewater to determine the feasibility of enriching N2O by in situ separation of dissolved gases. Gas separation across the membrane (GSM) had no obvious effect on chemical oxygen demand and nitrate removal, but N2O production efficiency (NPE) increased from 11.0 % to 82.6 %. Denitrifying bacteria with low N2O yield such as Thauera, Diaphorobacter, and Flavobacterium were eliminated, and Propioniclava (46.79 %) and Candidatus_Competibacter (22.53 %) became dominant. Metagenomic sequencing showed that the abundance of the nosZ gene encoding N2O reductase decreased from 17.3 % to 6.1 %. In conclusion, GSM prevented the further reduction of N2O, eliminating denitrifying bacteria carrying the nosZ gene, which the enriched Propioniclava likely lacked. This demonstrates that GSM is an alternative strategy for N2O enrichment through nitrate denitrification.

Metabolic Rates of Rainbow Trout Eggs in Reconstructed Salmonid Egg Pockets

In situ evaluations of the metabolic rates (i.e., respiration and excretion) of salmonid eggs are mostly indirect, focusing on the sampling of hyporheic water from wild or artificial nests. Comparatively, experimental studies carried out under controlled, laboratory conditions are less abundant due to methodological difficulties. This study presents a novel experimental setup aimed to address this issue and enable the measurement of oxygen and dissolved inorganic nitrogen fluxes in simulated rainbow trout (O. mykiss) egg pockets. The experimental setup consists of reconstructed egg pockets in cylindrical cores under flow-through conditions. Live and dead eyed-stage eggs were incubated in a natural, sterilised gravel substrate. Hyporheic water circulation was ensured using peristaltic pumps, with the possibility to collect and analyse inflowing and outflowing water for chemical analyses. Microcosm incubations, with closed respirometry of eggs in water alone, were also carried out in order to infer the importance of microbial respiration in the simulated egg pockets. The results show an increasing trend in oxygen demand, due to the development of biofilm in the reconstructed egg pockets and increased egg respiration rates. Moreover, egg pockets showed positive ammonium net fluxes connected with the advancing developmental egg stage, while nitrate removal peaked during the last phase of the experiment, mainly due to the formation of oxic-hypoxic interfaces, leading to couple nitrification–denitrification processes. The suggested approach enables to test a number of in situ situations, including the effects of extreme hydrological conditions, sediment clogging and sudden changes in water chemistry or temperature on the survival and metabolic performances of nests, at different egg development stages.

Self-cultivating anammox granules for enhancing wastewater nitrogen removal in nitrification–denitrification flocculent sludge system

The feasibility of self-cultivating anammox granules for enhancing wastewater nitrogen removal was investigated in a nitrification–denitrification flocculent sludge system. Desirable nitrogen removal efficiency of 84 ± 4 % was obtained for the influent carbon to nitrogen ratio of 1–1.3 (NH4+-N: 150–200 mg N/L) via alternate anaerobic/oxic/anoxic mode. Meanwhile, some red granular sludge was formed in the system. The abundance and activity of anaerobic ammonia oxidation bacteria (AnAOB) increased from ‘not detected’ in seed sludge to 0.57 % and 29.4 ± 0.7 mg N/(g mixed liquor volatile suspended solids·h) in granules, respectively, suggesting successful cultivation of anammox granules. Furthermore, some denitrifying bacteria with capability of partial denitrification were enriched, such as Candidatus Competibacter (2.45 %) and Thauera (5.75 %), which could cooperate with AnAOB, facilitating AnAOB enrichment. Anammox was dominant in nitrogen removal with the contribution to nitrogen removed above 68.8 ± 0.3 %. The strategy of self-cultivating anammox granules could promote the application of anammox.