Heterotrophic Nitrogen Research Articles

DOM and DON transformation in full-scale wastewater treatment plants: Comparison of autotrophic and heterotrophic nitrogen removal units

There is limited knowledge regarding the differences and relationships in molecular composition between dissolved organic matters (DOM) and dissolved organic nitrogen (DON) in autotrophic and heterotrophic nitrogen removal processes. In this study, the composition of DOM and DON during two full-scale wastewater treatment plants (WWTPs), an autotrophic nitrogen removal with partial nitrification-anammox process (PN/A) and a heterotrophic nitrogen removal with anaerobic/anoxic/oxic process (A2O), were systematically investigated by using ultraviolet–visible spectroscopy, Excitation-Emission Matrix (EEM) and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The results indicated that both PN/A and A2O resulted a decrease in molecular diversity and bio-availability of DOM. However, there were distinct variations in DON between the two processes. In PN/A, transformation of DOM and DON was found to be coupled, while there was a lack of correlation observed between DOM and DON in A2O. Pearson correlation analysis revealed that transformation of DOM and DON in WWTPs was associated with microbial roles. During biotreatment, PN/A exhibited a higher metabolic capacity for degrading high bio-available and high aromatic compounds compared to A2O. The autotrophic nitrogen removal unit in PN/A resulted in an increase in molecular weight (MW) of DON, as well as a decrease in bio-availability and unsaturation degree of DON. Therefore, autotrophic nitrogen removal unit may have a positive influence on control of eutrophication and disinfection by-products (DBPs) formation. This study improved the understanding of molecular composition and relationship between DOM and DON, as well as microbial roles on DON transformation within autotrophic and heterotrophic nitrogen removal processes.

Nitrogen-fixing sulfate reducing bacteria in shallow coastal sediments under simulated resuspension

Nitrogen (N2) fixation by heterotrophic non-cyanobacterial diazotrophs is common in marine deep-sea sediments. However, in shallow coastal areas, where resuspension of sediments is extensive, the magnitude of sediment-associated N2 fixation during resuspension is unknown. We examined nitrogen fixation in dark slurry incubations with sediments (0–5 cm and 0–10 cm depths) under simulated resuspension from five shallow stations (water depth <1 m) and an anoxic site in the Bay of Gdansk (109 m) in the Baltic Sea. Abiotic variables and the composition of nitrogen fixing organisms (diazotrophs) were measured at the study sites. To estimate the contribution of nitrogen fixing sulfate reducing bacteria to total nitrogen fixation, parallel incubations with sodium molybdate as inhibitor were performed. Our data show low but variable nitrogen fixation rates (n.d. - 23.7 nmol N g−1 d−1), promoted in small-grained sediments associated with increased organic carbon content and high nutrient concentrations in pore waters. Highest nitrogen fixation at the shallow sites was encountered in the upper 0–5 cm of the sediments while rates were negligible below. Sulfate reducing bacteria (e.g. Desulfobacterales and Desulfovibrionales) were responsible for most of the heterotrophic nitrogen fixation and appear as key players for pelagic N2 fixation during resuspension. Our study reveals an important sediment – water coupling, which may be accentuated by the increased storms and resuspension events predicted for the Baltic Sea region.

Capture of mammal excreta by Nepenthes is an effective heterotrophic nutrition strategy.

While isotopic enrichment of nitrogen (15N) and carbon (13C) is often used to determine whether carnivorous plant species capture and assimilate nutrients from supplemental sources such as invertebrate prey or mammal excreta (heterotrophic nutrition), little is known about how successful the different strategies deployed by carnivorous plants are at obtaining supplemental nutrition. The collection of mammalian faeces by Nepenthes (tropical pitcher plants) is the result of a highly specialized biological mutualism that results in heterotrophic nitrogen gain; however, it remains unknown how effective this strategy is in comparison to Nepenthes species not known to collect mammalian faeces. We examined how isotopic enrichment varied in the diverse genus Nepenthes, among species producing pitchers for invertebrate capture and species exhibiting mutualisms for the collection of mammal excreta. Enrichment factors were calculated from δ15N and δ13C values from eight Nepenthes species and naturally occurring hybrids along with co-occurring reference (non-carnivorous) plants from three mountain massifs in Borneo: Mount Kinabalu, Mount Tambuyukon and Mount Trus Madi. All Nepenthes examined, except N. edwardsiana, were significantly enriched in 15N compared to co-occurring non-carnivorous plants, and 15N enrichment was more than two-fold higher in species with adaptations for the collection of mammal excreta compared with other Nepenthes. The collection of mammal faeces clearly represents a highly effective strategy for heterotrophic nitrogen gain in Nepenthes. Species with adaptations for capturing mammal excreta occur exclusively at high elevation (i.e. are typically summit-occurring) where previous studies suggest invertebrate prey are less abundant and less frequently captured. As such, we propose this strategy may maximize nutritional return by specializing towards ensuring the collection and retention of few but higher-value N sources in environments where invertebrate prey may be scarce.

Symbiodiniaceae Are the First Site of Heterotrophic Nitrogen Assimilation in Reef-Building Corals.

ABSTRACTCoral reefs depend on the highly optimized mutualistic relationship between corals and Symbiodiniaceae dinoflagellates. Both partners exchange nutrients obtained through heterotrophy of the host and autotrophy of the symbionts. While heterotrophy helps corals withstand the harmful effects of seawater warming, the exchange of heterotrophic nutrients between the two partners is poorly understood. Here, we used compound-specific δ15N and δ13C of amino acids (δ15NAA and δ13CAA) and a 15N pulse-chase experiment with Artemia salina nauplii in two coral-dinoflagellate associations to trace the assimilation and allocation of heterotrophic nutrients within the partners. We observed that changes in the trophic position (TPGlx-Phe), δ15NAA, and δ13CAA with heterotrophy were holobiont-dependent. Furthermore, while TPGlx-Phe and δ15N of all AAs significantly increased with heterotrophy in the symbionts and host of Stylophora pistillata, only the δ15NAA of the symbionts changed in Turbinaria reniformis. Together with the pulse-chase experiment, the results suggested a direct transfer of heterotrophically acquired AAs to the symbionts of S. pistillata and a transfer of ammonium to the symbionts of T. reniformis. Overall, we demonstrated that heterotrophy underpinned the nutrition of Symbiodinaceae and possibly influenced their stress tolerance under changing environmental conditions.

Nitrogen content and C/N ratio in straw are the key to affect biological nitrogen fixation in a paddy field

Straw amendment can increase nitrogen fixation in paddy field, however, the efficiency of carbon sources with different biochemical properties to enhance N2 fixation and nitrogen fixation activity is still unclear. A 15N2-labelling system was used in the field environment to determine biological nitrogen fixation (BNF) under the addition of three kinds of straw. The nifH gene (DNA) and nifH RNA gene (cDNA) of soil were amplified by real-time fluorescent quantitative PCR. The diversity and community composition of nitrogen fixing microorganisms were studied by high-throughput sequencing. The study is expected to reveal how different carbon sources impact biological nitrogen fixation and its mechanism in the paddy field system. Results showed that the absolute abundance diazotrophs in the treatment of mature stage wheat straw (MWS) was 4.88 times as high as that in the CK treatment, but jointing stage wheat straw (JWS) and poplar branch (PB) did not induce any significant changes. Straw amendment had no impact on cyanobacteria abundance. The proportion of N2 fixation increased by MWS was 2.07 times, but which was much lower than the increase proportion of the heterotrophic diazotrophs, leading to a decrease of diazotrophic nitrogen fixation activity. Mature wheat straw addition increased biologically fixed nitrogen in paddy field by increasing the number of heterotrophic nitrogen fixing bacteria. The results indicated that to increase biological N2 fixation in paddy system, straws with low nitrogen content and high C/N ratio were recommended.

Biologically induced phosphate precipitation in heterotrophic nitrification processes of different microbial aggregates: Influences of nitrogen removal metabolisms and extracellular polymeric substances

Phosphorus (P) removal occurred in heterotrophic nitrification process, but its mechanism has not been fully explored. In this study, the P removal performances, pathways, and mechanisms in heterotrophic nitrification processes of different microbial aggregates (activated sludge and biofilm) were investigated. The results showed that the biofilm reactor had more efficient total nitrogen removal (98.65%) and phosphate removal (94.17%). Heterotrophic nitrification and denitrification processes generated alkalinity for biologically induced phosphate precipitation (BIPP), which contributed to 64.12%-78.81% of the overall P removal. The solid phase P content reached 48.03 mg/gSS with hydroxyapatite and calcium phosphate formation. The study clarified that biofilm was beneficial to BIPP because of the nitrogen removal metabolism and extracellular polymeric substance (EPS). Heterotrophic nitrogen removal metabolism was the driving force of BIPP, while EPS with abundant carboxyl and amide groups promoted the precipitation. The study would provide new insights into simultaneous nutrients removal and P recovery from wastewater.

Microorganisms as bio-filters to mitigate greenhouse gas emissions from high-altitude permafrost revealed by nanopore-based metagenomics.

The distinct climatic and geographical conditions make high-altitude permafrost on the Tibetan Plateau suffer more severe degradation than polar permafrost. However, the microbial responses associated with greenhouse gas production in thawing permafrost remain obscured. Here we applied nanopore-based long-read metagenomics and high-throughput RNA-seq to explore microbial functional activities within the freeze-thaw cycle in the active layers of permafrost at the Qilian Mountain. A bioinformatic framework was established to facilitate phylogenetic and functional annotation of the unassembled nanopore metagenome. By deploying this strategy, 42% more genera could be detected and 58% more genes were annotated to nitrogen and methane cycle. With the aid of such enlarged resolution, we observed vigorous aerobic methane oxidation by Methylomonas, which could serve as a bio-filter to mitigate CH4 emissions from permafrost. Such filtering effect could be further consolidated by both on-site gas phase measurement and incubation experiment that CO2 was the major form of carbon released from permafrost. Despite the increased transcriptional activities of aceticlastic methanogenesis pathways in the thawed permafrost active layer, CH4 generated during the thawing process could be effectively consumed by the microbiome. Additionally, the nitrogen metabolism in permafrost tends to be a closed cycle and active N2O consumption by the topsoil community was detected in the near-surface gas phase. Our findings reveal that although the increased thawed state facilitated the heterotrophic nitrogen and methane metabolism, effective microbial methane oxidation in the active layer could serve as a bio-filter to relieve the overall warming potentials of greenhouse gas emitted from thawed permafrost.

High Abundance of Thaumarchaeota Found in Deep Metamorphic Subsurface in Eastern China.

Members of the Thaumarchaeota phylum play a key role in nitrogen cycling and are prevalent in a variety of environments including soil, sediment, and seawater. However, few studies have shown the presence of Thaumarchaeota in the terrestrial deep subsurface. Using high-throughput 16S rRNA gene sequencing, this study presents evidence for the high relative abundance of Thaumarchaeota in a biofilm sample collected from the well of Chinese Continental Scientific Drilling at a depth of 2000 m. Phylogenetic analysis showed a close relationship of these thaumarchaeotal sequences with known ammonia-oxidizing archaea (AOA) isolates, suggesting the presence of AOA in the deep metamorphic environment of eastern China which is believed to be oxic. Based on fluid geochemistry and FAProTax functional prediction, a pathway of nitrogen cycling is proposed. Firstly, heterotrophic nitrogen fixation is executed by diazotrophic bacteria coupled with methane oxidation. Then, ammonia is oxidized to nitrite by AOA, and nitrite is further oxidized to nitrate by bacteria within the phylum Nitrospirae. Denitrification and anaerobic ammonia oxidation occur slowly, leading to nitrate accumulation in the subsurface. With respect to biogeochemistry, the reaction between downward diffusing O2 and upward diffusing CH4 potentially fuels the ecosystem with a high relative abundance of Thaumarchaeota.

Superior nitrogen removal and sludge reduction in a suspended sludge system with in-situ enriching anammox bacteria for real sewage treatment

Mainstream partial anammox provides a cost- and energy-efficient alternative for wastewater treatment. This study provided a new strategy to achieve mainstream partial anammox in a single-stage suspended sludge system. The novel method coupling external excess sludge fermentation with simultaneous partial nitritation-anammox-denitrification process (SF-SPNAD) was established for 202 days in an anaerobic-aerobic-anoxic sequencing batch reactor (AOA-SBR) with real sewage and actual sludge fermentation products. Under the condition of low DO (0.6 ± 0.2 mg/L), short oxic and long anoxic hydraulic retention time (HRToxic = 6.5 h, HRTanoxic = 8 h), the average total inorganic nitrogen (TIN) concentration in the influent and effluent during 110-day operation were 61.0 and 3.4 mg/L, respectively, and the TIN removal efficiency was 94.56%. Under the inhibitory effect of continuous sludge fermentation products addition, nitrite accumulation ratio reached 99.1% and the external sludge reduction ratio reached 38.75%. 15N-stable isotope tracing tests showed the great potential of nitrogen removal by anammox pathway in the system. High-throughput sequencing confirmed that CandidatusBrocadia (not detected to 0.50%) and CandidatusKuenenia (not detected to 0.06%) were successfully in-situ enriched. Nitrogen conversion pathways based on stoichiometry and cycle tests show that 34.69% of the TIN removal was obtained by simultaneous nitritation denitrification and anammox under oxic stage and 35.21% of the TIN removal was carried out by anammox under anoxic stage. Overall, the SF-SPNAD process provides a new possibility for coupling autotrophic and heterotrophic nitrogen removal with excess sludge utilization.

Heterotrophic nitrogen fixation in response to nitrate loading and sediment organic matter in an emerging coastal deltaic floodplain within the Mississippi River Delta plain

AbstractIncreasing nitrate (NO3−) loading in rivers due to agricultural fertilization alters benthic nitrogen (N) cycling and shifts coastal wetlands from being a net source to net sink of reactive N. Heterotrophic N2 fixation that converts N2 to reactive N is often assumed negligible in eutrophic ecosystems and excluded in coastal N budget evaluations. We investigated N2 fixation and denitrification in response to increasing NO3− loading (0, 10, and 100 μM) and sediment organic matter (OMsediment) concentrations in the emerging Wax Lake Delta. Continuous flow‐through incubations with 30N2 addition was applied to measure N2 fixation. The variation of N2 fixation rates from 0 to 437 μmol N m−2 h−1 among different NO3− and OMsediment concentrations were comparable to the estimated denitrification rates of 141–377 μmol N m−2 h−1. Increasing overlying NO3− concentrations reduced N2 fixation rates and facilitated denitrification rates at each OMsediment concentration. However, 100 μM of overlying NO3− did not thoroughly inhibit N2 fixation rates in sites with intermediate and higher OMsediment concentrations (189 and 99 μmol N m−2 h−1, respectively). Both N2 fixation and denitrification increased with increasing OMsediment concentrations, but the relative importance of these processes was impacted mostly by overlying NO3− concentration as increasing NO3− switched the dominance of N2 fixation to denitrification in benthic N cycling. This study highlights the importance of heterotrophic N2 fixation in coastal deltaic floodplains and emphasizes the necessity of including N2 fixation quantification in coastal N budget evaluation, not only in oligotrophic environment but also in eutrophic environment.

Energetic supply regulates heterotrophic nitrogen fixation along a glacial chronosequence

Ice-free areas are expanding on the Tibetan Plateau and in its surroundings, and these recently exposed lands are typically deficient in nitrogen (N). Free-living N fixation (FLNF) represents a vital source of N input, especially when symbiotic N-fixing plants are absent. However, we know little about how FLNF changes with ecosystem succession after glacier retreat. In 2018, we measured the potential rates of FLNF in soil profiles at four sites (exposed since 2012, 1980, 1970 and 1930, respectively) along the Hailuogou glacier chronosequence and at a reference site with well-developed Podosols. We also tested the role of nifH gene abundance and resource availability in regulating FLNF rates. The results showed that photosynthetic cyanobacteria likely contributed the majority of FLNF in the biological soil crusts (exposed since 2012), while heterotrophic FLNF may prevail at later successional stages with closed canopies. FLNF rates were the highest in the litter layer of the soil profiles and decreased with soil depth. In the litter layer, FLNF rates were lower at the site dominated by N-fixing shrubs (exposed since 1980) than at the older sites dominated by non-N-fixing trees (exposed since 1970 and 1930, and the reference site). Carbon (C) availability was the major factor explaining variation in heterotrophic FLNF rates measured in the lab without light, while the effect of nifH gene abundance was nonsignificant. Overall, our results highlighted the energetic control on heterotrophic FLNF rates in relatively young ecosystems, which likely resulted in the poor relationship between nifH gene abundance and heterotrophic FLNF rates. The findings will help in understanding and modeling the coupled C and N cycles in glacier forefield ecosystems which are being exposed at accelerated rates.

Heterotrophic Nitrogen Fixation at the Hyper-Eutrophic Qishon River and Estuary System.

Planktonic heterotrophic diazotrophs (N2-fixers) are widely distributed in marine and freshwater systems, yet limited information is available on their activity, especially in environments with adverse conditions for diazotrophy (e.g., N-rich and oxygenated). Here, we followed the localization and activity of heterotrophic diazotrophs in the hyper-eutrophic N-rich Qishon River—an environment previously considered to be unfavorable for diazotrophy. Our results indicate high heterotrophic N2 fixation rates (up to 6.9 nmol N L–1 d–1), which were approximately three fold higher at an upstream location (freshwater) compared to an estuary (brackish) site. Further, active heterotrophic diazotrophs were capture associated with free-floating aggregates by a newly developed immunolocalization approach. These findings provide new insights on the activity of heterotrophic diazotrophs on aggregates in environments previously considered with adverse conditions for diazotrophy. Moreover, these new insights may be applicable to other aquatic regimes worldwide with similar N-rich/oxygenated conditions that should potentially inhibit N2 fixation.

Long-term stable and energy-neutral mixed biofilm electrode for complete nitrogen removal from high-salinity wastewater: Mechanism and microbial community

The steady mixed biofilm electrode (MBE) was investigated for the removal of nitrogen from mustard tuber wastewater. Results showed that complete nitrogen removal occurred over a wide initial chemical oxygen demand (COD)/total nitrogen (TN) ratio ranging from 2.8 to 9.8 using MBE. MBE revealed broad-spectrum applicability for the treatment of high-salinity wastewater containing different forms of nitrogen. Bio-electrochemical process, in-situ heterotrophic nitrogen reduction, ammonia stripping, nitrogen assimilation, and endogenous denitrification coexisted for the removal of nitrogen. Batch activity tests and functional microorganism analysis confirmed that autotrophic/heterotrophic nitrification, anoxic/aerobic denitrification, and nitrogen bio-electrochemical reduction cooperated to achieve efficient nitrogen conversion. More importantly, the analysis of the preliminary energy balance demonstrated that MBE was self-sustaining. The long-term operation stability of MBE was of great importance for its practical application. The results provided herein offer new insights into bioelectrochemical nitrogen removal and resource treatment of high-salinity wastewater.

Screening Heterotrophic Ammonia Removal and Aerobic Denitrifying Bacteria from Wastewater of Ammonia Production Units of a Petrochemical Industry.

Petrochemical wastewaters have high ammonium contents that can cause serious environmental pollutions. Among different treatment methods, biological ammonia removal using bacteria has been attracted more attention due to their environmental friendly and high efficiency. The aim of the present study was to isolate heterotrophic nitrogen removal and aerobic denitrifying bacteria from petrochemical wastewater to be used for bioremediation. Wastewater and sludge samples were prepared and their chemical parameters were analyzed. Bacterial isolation was done through enrichment in mineral salt medium and basal salt medium and identified through 16S rRNA gene sequencing. The ability of isolates was evaluated at different concentrations of ammonia, nitrate, and carbon sources and at different time intervals. As a result, ten bacterial isolates were obtained that 5 strains were identified as heterotrophic nitrogen removal bacteria and 5 strains were able to do aerobic denitrification. From these, Pseudomonas guguanensis strain 4-n-1 with 93.2% ammonia removal during 24h at 40mg/L ammonium sulfate concentration and Pseudomonas guariconensis strain 5-d-1 with 89.2% denitrification during 24h at 100mg/L nitrate concentration were the best isolates. Finally, based on the efficiency of these isolates it can be concluded that effective ammonia removal can be achieved using these species as consortium and simultaneous heterotrophic nitrogen removal with aerobic denitrification is suitable method for efficient ammonium removal from petrochemical wastewater and reduction of ammonia concentration.

Development of a single-stage mainstream anammox process using a sponge-bed trickling filter

ABSTRACT Anaerobic ammonia oxidation to nitrogen gas using nitrite as the electron acceptor (anammox process) is considered a cost-effective solution for nitrogen removal after an anaerobic pre-treatment process. In this study, we conducted a laboratory-scale experiment to develop a single-stage partial nitritation–anammox process in a sponge-based trickling filter (STF) reactor, inoculated with anammox sludge, simulating the treatment of anaerobically pretreated concentrated domestic sewage without mechanical oxygen control. The influent ammonia concentration was 100 mg-N·L−1. The KLa of the STF reactor was higher than those observed for conventional activated sludge processes. The STF reactor performed at 89.8 ± 8.2% and 42.7 ± 16.9% ammonia and TN removal efficiency, respectively, with a nitrogen loading rate of 0.55 ± 0.20 kg-N·m−3·day−1 calculated based on sponge volume. Microbial community analysis of the STF-retained sludge indicated that both autotrophic and heterotrophic nitrogen removal occurred in the reactor.

Heterotrophic nitrogen removal by a newly-isolated microorganism, Oligella sp. XS68

A new bacterium capable of heterotrophic nitrogen removal was isolated and identified as Oligella sp. XS68. The strain exhibited efficient heterotrophic nitrogen removal capabilities, with a low accumulation of nitrification products. Single-factor experiments indicated that efficient nitrogen removal and growth of the strain XS68 occurred with sodium succinate as the carbon source, C/N ratio 15, pH 6.0?9.0, temperature 30?37 ?C, and shaking speed 160?200 rpm. The removal efficiency of ammonium nitrogen could achieve 98 % within 96 h when the initial nitrogen concentration was 421.3 mg L-1. These findings demonstrate that XS68 is a promising candidate for nitrogen removal in wastewater treatments.

A quantitative model of nitrogen fixation in the presence of ammonium.

Nitrogen fixation provides bioavailable nitrogen, supporting global ecosystems and influencing global cycles of other elements. It provides an additional source of nitrogen to organisms at a cost of lower growth efficiency, largely due to respiratory control of intra-cellular oxygen. Nitrogen-fixing bacteria can, however, utilize both dinitrogen gas and fixed nitrogen, decreasing energetic costs. Here we present an idealized metabolic model of the heterotrophic nitrogen fixer Azotobacter vinelandii which, constrained by laboratory data, provides quantitative predictions for conditions under which the organism uses either ammonium or nitrogen fixation, or both, as a function of the relative supply rates of carbohydrate, fixed nitrogen as well as the ambient oxygen concentration. The model reveals that the organism respires carbohydrate in excess of energetic requirements even when nitrogen fixation is inhibited and respiratory protection is not essential. The use of multiple nitrogen source expands the potential niche and range for nitrogen fixation. The model provides a quantitative framework which can be employed in ecosystem and biogeochemistry models.

Plant-soil feedbacks on free-living nitrogen fixation over geological time.

Free-living heterotrophic nitrogen fixation (FNF) is a widespread nitrogen input pathway in terrestrial ecosystems. However, questions remain over the relative influence of co-occurring controls on patterns of heterotrophic FNF activity, especially across generalized stages of primary succession, from biomass accumulation to retrogressive phases. Here, we experimentally test two alternative hypotheses regarding FNF rates during ecosystem development: (H1) site (i.e., changes in soil fertility during succession) is the primary driver of leaf-litter FNF rates, vs. (H2) leaf-litter chemistry is the primary determinant of FNF activity across a broad range of ecosystem conditions. We evaluated these hypotheses across a well-studied soil chronosequence in California (i.e., the Ecological Staircase), which spans ~1million years of ecosystem development and displays extreme ranges in plant-soil nutrient conditions, culminating in the nutrient depleted and stunted Pygmy forest. Across this successional gradient, we implemented a reciprocal leaf-litter transplant and a common garden litter bag decomposition experiment with senesced needles of Pinus muricata. Our results support H1: rates of FNF were similar for all leaf-litter types decomposed at the same site regardless of initial leaf-litter C and nutrient contents. FNF rates sharply declined from the maximal to retrogressive stage of succession. Trends in P dynamics during decomposition suggest an important role of P in regulating FNF. For example, P.muricata litter collected from the infertile Pygmy site displayed substantially higher FNF rates when decomposed at the fertile site, in part by immobilizing significant quantities of P from the soil at the fertile site. Conversely, P.muricata litter collected from the fertile site decomposed more slowly at the Pygmy site, with concomitant declines in FNF rates that matched those of Pygmy site litter decomposed insitu. These results are consistent with the idea that, over millennia, long-term declines in P availability feedback to constrain FNF rates, in part explaining the emergence of extremely nutrient-poor and retrograded ecosystems.

High-efficiency inorganic nitrogen removal by newly isolated Pannonibacter phragmitetus B1

An aerobic heterotrophic nitrogen removal bacterium strain, B1, was isolated from aquaculture water and identified as Pannonibacter phragmitetus (99% similarity) by 16S rRNA sequencing analysis. When ammonium, nitrite or nitrate was the sole nitrogen source, with an initial nitrogen concentration of 14 mg/L, the nitrogen removal efficiencies were 98.66%, 99.96% and 98.73%, respectively, and the corresponding maximum removal rates reached as high as 1.16, 0.77 and 0.81 mg/L/h, respectively. In the presence of NH4+-N, the removal efficiency of 56 mg/L NO2−-N within 27 h increased by 83.50%, and the corresponding removal rate reached as high as 1.72 mg/L/h. Additionally, different carbon sources (dl-malic acid, sucrose, sodium citrate, and glucose) could be utilized in nitrogen removal. Sequence amplification indicates that the denitrification genes nirK, norB and narG are present in strain B1. All results demonstrate that strain B1 has high promise for future applications of removing inorganic nitrogen from wastewater.

Novel heterotrophic nitrogen removal and assimilation characteristic of the newly isolated bacterium Pseudomonas stutzeri AD-1

AD-1, an aerobic denitrifier, was isolated from activated sludge and identified as Pseudomonas stutzeri. AD-1 completely removed NO3- or NO2- and removed 99.5% of NH4+ during individual culturing in a broth medium with an initial nitrogen concentration of approximately 50mgL-1. Results showed that larger amounts of nitrogen were removed through assimilation by the bacteria. And when NH4+ was used as the sole nitrogen source in the culture medium, neither NO2- nor NO3- was detected, thus indicating that AD-1 may not be a heterotrophic nitrifier. Only trace amount of N2O was detected during the denitrification process. Single factor experiments indicated that the optimal culture conditions for AD-1 were: a carbon-nitrogen ratio (C/N) of 15, a temperature of 25°C and sodium succinate or glucose as a carbon source. In conclusion, due to the ability of AD-1 to utilize nitrogen of different forms with high efficiencies for its growth while producing only trace emissions of N2O, the bacterium had outstanding potential to use in the bioremediation of high-nitrogen-containing wastewaters. Meanwhile, it may also be a proper candidate for biotreatment of high concentration organic wastewater.