Nitrogen Metabolic Pathways Research Articles

Relationships between high-concentration toxic metals in sediment and evolution of microbial community structure and carbon-nitrogen metabolism functions under long-term stress perspective.

Microorganisms are highly sensitive to toxic metal pollution and play an important role in the material cycling and energy flow of the water ecosystem. Herein, 13 sediment samples from Junchong Reservoir (Guangxi Province, China) were collected in December 2021. The spatial distribution of pollution levels for toxic metals and the effects of toxic metals on the composition, functional characteristics, and metabolism of microorganisms were investigated. The results demonstrated that the area is a proximate area to industrial zones with severity of toxic metal pollution. Their mean concentrations of As, Cu, Zn, and Pb were up to 128.79mg/kg, 57.62mg/kg, 594.77mg/kg, and 97.12mg/kg respectively. There was a strong correlation between As, Cu, Zn, and Pb, with the highest correlation coefficient reaching 0.94. As the level of toxic metal pollution increases, the diversity and abundance of microorganisms gradually decrease. Compared to those with lower pollution levels, the Shannon index in regions with higher pollution levels decreases by up to 0.373, and the Chao index decreases by up to 143.507. However, the relative abundance of Bacteroidota, Patescibacteria, and Chloroflexi increased by 23%, 20%, and 5%, respectively, indicating their higher adaptability to toxic metals. Furthermore, microbial carbon and nitrogen metabolism were also affected by the presence of toxic metals. FAPROTAX analysis demonstrated an abundant reduction of ecologically functional groups associated with carbon and nitrogen transformations under high toxic metal pollution levels. KEGG pathway analysis indicated that carbon fixation and nitrogen metabolism pathways were inhibited with increasing toxic metal concentrations. These findings would contribute to a better understanding of the effects of toxic metal pollution on sediment microbial communities and function, shedding light on the ecological consequences of toxic metal contamination.

Biomineralized manganese oxide mediated nitrogen-contained wastewater treatment

In recent years, manganese (Mn) has emerged as an accelerator for nitrogen metabolism. However, the bioactivity of manganese is limited by the restricted contact between microbes and manganese minerals in the solid phase and by the toxicity of manganese to microbes. To enhance the bioactivity of solid-phase manganese, biomineralized manganese oxide (MnOx) modified by Lactobacillus was introduced. Nitrogen removal performance have confirmed the effective role of biomineralized MnOx in accelerating the removal of total inorganic nitrogen (TIN). Metagenomic analysis has confirmed the enhancement of the nitrogen metabolic pathway and microbial extracellular electron transfer (MEET) in biomineralized MnOx treatment group (BIOA group). Additionally, the enrichment of manganese oxidation and denitrification genus indicates a coupling between nitrogen metabolism and manganese metabolism. One point of views is that biomineralized MnOx-mediated nitrogen transformation processes could serve as a substitute for traditional nitrogen removal processes.

Effects of mycorrhizal symbiosis and Ulva lactuca seaweed extract on growth, carbon/nitrogen metabolism, and antioxidant response in cadmium-stressed sorghum plant.

In our study on the effect of cadmium (Cd) toxicity (200µM) on the growth of Sorghum bicolor (L.) Moench plants, cultivated with arbuscular mycorrhizal fungi (AMF) (Glomus intraradices) and/or under seaweed treatment (3% Ulva lactuca extract) (U. lactuca), we found that AMF increased the tolerance of sorghum to cadmium stress, either alone or in combination with the seaweed treatment. Morphological parameters were higher in these two culture conditions, with increased chlorophyll content. AMF reduced Cd accumulation in roots and inhibited its translocation to the aerial part, while seaweed treatment alone significantly increased Cd accumulation in leaves and roots without affecting plant growth compared to stressed witnesses. Treatment with AMF and/or U. lactuca attenuated oxidative stress, measured by activation of superoxide dismutase, and resulted in a significant decrease in malondialdehyde and superoxide ions (O2-) in treated plants. Furthermore, it induced significant alterations in carbon and nitrogen metabolic pathways, with a significant increase in the activity of enzymes such as glutamine synthetase, glutamate synthase (GOGAT), glutamate dehydrogenase, phosphoenolpyruvate carboxylase, aspartate aminotransferase and isocitrate dehydrogenase in the leaves of each treated plant. These results confirm that AMF, U. lactuca algae extract and their combination can improve the biochemical parameters of sorghum under Cd stress, through modification of the antioxidant response on one hand, and improved nitrogen absorption and assimilation efficiency on the other.

Iron-enhanced microscale laboratory aerated filters in the treatment of artificial mariculture wastewater: A study on nitrogen removal performance and the impact on microbial community structure

This study investigates the nitrogen removal efficacy and microbial community dynamics in seawater aquaculture effluent treatment using three different substrate combinations of microscale laboratory aerated filters (MFs) — MF1 (LECA), MF2 (LECA/Fe–C), and MF3 (LECA/Pyrite). The findings indicated that the COD removal exceeded 95% across all MFs, with higher removal efficiencies in MF2 and MF3. In terms of nitrogen removal performance, MF2 exhibited the highest average nitrogen removal of 93.17%, achieving a 12.35% and 3.56% increase compared to MF1 (80.82%) and MF3 (89.61%), respectively. High-throughput sequencing analysis revealed that the Fe–C substrate significantly enhanced the diversity of the microbial community. Notably, in MF2, the salinophilic denitrifying bacterium Halomonas was significantly enriched, accounting for 42.6% of the total microbial community, which was beneficial for nitrogen removal. Moreover, an in-depth analysis of nitrogen metabolic pathways and microbial enzymes indicated that MF2 and MF3 possessed a high abundance of nitrification and denitrification enzymes, related to the high removal rates of NH4+-N and NO3−-N. Therefore, the combination of LECA with iron-based materials significantly enhances the nitrogen removal efficiency from mariculture wastewater.

Transcript Analysis Reveals Positive Regulation of CA12g04950 on Carotenoids of Pigment Pepper Fruit under Nitrogen Reduction

This study investigates the relationship between nitrogen fertilization and pepper fruit color by employing five different nitrogen treatments (N1: 750 kg/hm2, N2: 562.5 kg/hm2, N3: 375 kg/hm2, N4: 187.5, and N0: 0 kg/hm2). Fruits were harvested at 30 (S1: green ripening stage), 45 (S2: color transition stage), and 60 days (S3: red ripening stage) after flowering. Subsequently, pigment content, carotenoid component content, carotenoid enzyme activity, and transcriptome sequence were analyzed, and CA12g04950 function was validated through virus-induced gene silencing (VIGS). The results indicate that a reduction in nitrogen application led to an earlier onset of fruit color breakdown, and increased the contents of total carotenoid, capsanthin, phytoene and PSY (phytoene synthase) activity, LCYB (lycopene β-cyclase) activity and CCS (capsanthin/capsorubin synthase) activity. The analysis of different expression genes indicated that the most differently expressed genes were enriched in the N1 vs. N4 comparison, with 18 genes involved in carotenoid metabolism and 16 genes involved in nitrogen metabolism. Most DE genes were enriched in the pathways of photosynthesis, porphyrin, carotenoid biosynthesis, seleno-compounds, and nitrogen metabolism. There were numerous differential transcription factor families, including ERF, bHLH, MYB, C2H2, and NAC. Pearson correlation analysis revealed a significant positive correlation between CA12g04950 expression and 11 carotenoid genes in the N4 treatment. Subsequent silencing of CA12g04950 using VIGS resulted in delayed color ripening while a significant decrease in total carotenoid content and the expression levels of carotenoid genes. In conclusion, nitrogen reduction led to an increase in carotenoid content in pigment pepper fruits. Furthermore, under nitrogen reduction, CA12g04950 positively influenced the redness of the fruits.

Identification of key genes and immune infiltration in osteoarthritis through analysis of zinc metabolism-related genes

BackgroundOsteoarthritis (OA) represents a prominent etiology of considerable pain and disability, and conventional imaging methods lack sensitivity in diagnosing certain types of OA. Therefore, there is a need to identify highly sensitive and efficient biomarkers for OA diagnosis. Zinc ions feature in the pathogenesis of OA. This work aimed to investugate the role of zinc metabolism-related genes (ZMRGs) in OA and the diagnostic characteristics of key genes.MethodsWe obtained datasets GSE169077 and GSE55235 from the Gene Expression Omnibus (GEO) and obtained ZMRGs from MSigDB. Differential expression analysis was conducted on the GSE169077 dataset using the limma R package to identify differentially expressed genes (DEGs), and the intersection of DEGs and ZMRGs yielded zinc metabolism differential expression-related genes (ZMRGs-DEGs). The clusterProfiler R package was employed for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of ZMRGs-DEGs. Potential small molecule drugs were predicted using the CMap database, and immune cell infiltration and function in OA individuals were analyzed using the ssGSEA method. Protein-protein interaction (PPI) networks were constructed to detect Hub genes among ZMRGs-DEGs. Hub gene expression levels were analyzed in the GSE169077 and GSE55235 datasets, and their diagnostic characteristics were assessed using receiver operating characteristic (ROC) curves. The gene-miRNA interaction network of Hub genes was explored using the gene-miRNA interaction network website.ResultsWe identified 842 DEGs in the GSE169077 dataset, and their intersection with ZMRGs resulted in 46 ZMRGs-DEGs. ZMRGs-DEGs were primarily enriched in functions such as collagen catabolic processes, extracellular matrix organization, metallopeptidase activity, and pathways like the IL-17 signaling pathway, Nitrogen metabolism, and Relaxin signaling pathway. Ten potential small-molecule drugs were predicted using the CMap database. OA patients exhibited distinct immune cell abundance and function compared to healthy individuals. We identified 4 Hub genes (MMP2, MMP3, MMP9, MMP13) through the PPI network, which were highly expressed in OA and demonstrated good diagnostic performance. Furthermore, two closely related miRNAs for each of the 4 Hub genes were identified.Conclusion4 Hub genes were identified as potential diagnostic biomarkers and therapeutic targets for OA.

Involvement of functional metabolism promotes the enrichment of antibiotic resistome in drinking water: Based on the PICRUSt2 functional prediction

Biofilters are the important source and sink of antibiotic resistance genes (ARGs) and antibiotic resistance bacteria (ARB) in the drinking water. Current studies generally ascribed the prevalence of BAR in biofilter from the perspective of gene behavior, i.e. horizontal gene transfer (HGT), little attentions have been paid on the ARGs carrier- ARB. In this study, we proposed the hypothesis that ARB participating in pollutant metabolism processes and becoming dominant is an important way for the enrichment of ARGs. To verify this, the antibiotic resistome and bacterial functional metabolic pathways of a sand filter was profiled using heterotrophic bacterial plate counting method (HPC), high-throughput qPCR, Illumina Hiseq sequencing and PICRUSt2 functional prediction. The results illustrated a significant leakage of ARB in the effluent of the sand filter with an average absolute abundance of approximately 102–103 CFU/mL. Further contribution analysis revealed that the dominant genera, such as Acinetobacter spp., Aeromonas spp., Elizabethkingia spp., and Bacillus spp., were primary ARGs hosts, conferring resistance to multiple antibiotics including sulfamethoxazole, tetracycline and β-lactams. Notably, these ARGs hosts were involved in nitrogen metabolism, including extracellular nitrate/nitrite transport and nitrite reduction, which are crucial in nitrification and denitrification in biofilters. For example, Acinetobacter spp., the dominant bacteria in the filter (relative abundance 69.97 %), contributed the majority of ARGs and 53.79 % of nitrite reduction function. That is, ARB can predominate by participating in the nitrogen metabolism pathways, facilitating the enrichment of ARGs. These findings provide insights into the stable presence of ARGs in biofilters from a functional metabolism perspective, offering a significant supplementary to the mechanisms of the emergence, maintenance, and transmission of BARin drinking water.

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.

Nitrogen metabolism pathways and functional microorganisms in typical karst wetlands.

Aha Lake artificial reservoir wetland, Niangniang Mountain karst mountain wetland, and Caohai plateau lake wetland are typical karst wetlands in Guizhou Province with unique topography and geomorphic features. They were selected as research objects in this study to explore microorganisms and functional genes in nitrogen metabolism adopting macro-genome sequencing technology. It was found that Proteobacteria, Actinobacteria, and Acidobacteria were the dominant phyla in nitrogen metabolism in these three wetlands, similar to previous studies. However, at the genus level, there was a significant difference, with the dominant bacteria being Bradyrhizobium, Methylocystis, and Anaeromyxobacter. Six nitrogen metabolism pathways, including nitrogen fixation, nitrification, denitrification, dissimilatory nitrate reduction, assimilatory nitrate reduction, and complete nitrification, comammox, were revealed, but anaerobic ammonia oxidation genes were not detected. Nitrogen metabolism microorganisms and pathways were more affected by SOM, pH, NO3-, and EC in karst wetlands. This study further discussed microorganisms and functions of nitrogen metabolism in karst wetlands, which was of great significance to nitrogen cycles of karst wetland ecosystems.

Microbiome comparisons reveal the core microorganisms and key nitrogen metabolic pathway involved in industrial nitrogen fertilizer wastewater treatment plants

Microbiomes from municipal wastewater treatment plants (WWTPs) have been widely investigated, but less attention has been paid to those in the industrial WWTPs. To compare the microbiomes involved in nitrogen fertilizer wastewater treatment, four WWTPs with different concentrations and types of pollutants, processes, and geographic locations were chosen. Diversity results revealed that microbial community was similar within the same WWTP, but different among the four WWTPs. CPCoA further demonstrated these differences (P = 0.022), implying that the diverse manufactured products acted as electron donors to create distinct ecological niches for denitrification. However, bacterial composition and functional profile were similar among four WWTPs (P > 0.05), and their abundance differed and fluctuated. More than 80 % of the dominant genera detected were present in all four WWTPs. And the core bacteria identified were Hyphomicrobium, Acinetobacter, Pedomicrobium, Methyloversatilis, Gp16, Thauera, and Moorella respectively, accounting for 19.524 % of total bacterial abundance. Forty-five nitrogen metabolism genes were active in four nitrogen cycle pathways (nitrification, assimilatory nitrate reduction, dissimilatory nitrate reduction, denitrification). The key genes were amoC (0.30 %), nasA (5.35 %), nirA (2.17 %), nirB (6.04 %), nirD (5.82 %), narG (3.87 %), narH (4.65 %), nirK (3.90 %), norB (3.32 %), nosZ (2.80 %). Hyphomicrobium, Thauera, Afipia, and Paracocccus were the hosts of denitrification genes, synergistically treating the nitrogen fertilizer wastewater to meet the discharge standard. These results suggested that microbial community structure in the nitrogen fertilizer industry is similar, helping further the understanding of the microbiota at an industry level though more samples are required to verify this.

Insights into the response of nitrogen metabolism to sulfamethoxazole contamination in constructed wetlands with varied substrates

This study conducted an analysis of the variations in nitrogen metabolism pathways within constructed wetlands (CWs) using zeolite (CW-Z), ceramsite (CW-C), and lava (CW-L) under high concentration sulfamethoxazole (SMX) stress. The introduction of SMX hindered the formation of hydrogen bonds on the substrate surfaces; however, these surfaces still maintained a dense and thick biofilm. CW-Z exhibited superior removal efficiencies for ammonium nitrogen (NH4+-N) and nitrate nitrogen (NO3–-N) compared to CW-C and CW-L, with removal rates of 92.54 ± 2.88 % and 89.39 ± 6.74 %, respectively. Interestingly, the proportion of genes involved in nitrification, denitrification and nitrate reduction genes in CW-C (36.05 %) were higher than that in CW-C (29.81 %) and CW-L (29.70 %) but the interactions among nitrogen functional bacteria in CW-Z were much more complex. Further analysis of the nitrogen metabolism pathway indicated that under CW-Z enhanced dissimilatory nitrate reduction SMX stress, while CW-L enhanced assimilatory nitrate reduction process compared to CW-C.

Effect of nitrogen reduction by chemical fertilization with green manure (Vicia sativa L.) on soil microbial community, nitrogen metabolism and and yield of Uncaria rhynchophylla by metagenomics.

Uncaria rhynchophylla is an important herbal medicine, and the predominant issues affecting its cultivation include a single method of fertilizer application and inappropriate chemical fertilizer application. To reduce the use of inorganic nitrogen fertilization and increase the yield of Uncaria rhynchophylla, field experiments in 2020-2021 were conducted. The experimental treatments included the following categories: S1, no fertilization; S2, application of chemical NPK fertilizer; and S3-S6, application of chemical fertilizers and green manures, featuring nitrogen fertilizers reductions of 0%, 15%, 30%, and 45%, respectively. The results showed that a moderate application of nitrogen fertilizer when combined with green manure, can help alleviate soil acidification and increase urease activity. Specifically, the treatment with green manure provided in a 14.71-66.67% increase in urease activity compared to S2. Metagenomics sequencing results showed a decrease in diversity in S3, S4, S5, and S6 compared to S2, but the application of chemical fertilizer with green manure promoted an increase in the relative abundance of Acidobacteria and Chloroflexi. In addition, the nitrification pathway displayed a progressive augmentation in tandem with the reduction in nitrogen fertilizer and application of green manure, reaching its zenith at S5. Conversely, other nitrogen metabolism pathways showed a decline in correlation with diminishing nitrogen fertilizer dosages. The rest of the treatments showed an increase in yield in comparison to S1, S5 showing significant differences (p < 0.05). In summary, although S2 demonstrate the ability to enhance soil microbial diversity, it is important to consider the long-term ecological impacts, and S5 may be a better choice.

Metabolic profiling of leaves of four &lt;i&gt;Ranunculus&lt;/i&gt; species

BACKGROUND: Plant ability to survive oxygen deficiency is associated with the presence of various adaptations, majority of which are mediated by significant changes of metabolism. These alterations allow resistant wetland plants to grow even in an oxygen-depleted environment.&#x0D; AIM: To compare metabolic profiles of the leaves of the wetland species Ranunculus lingua, R. repens and R. sceleratus, and the mesophyte species R. acris growing in their natural habitat in order to identify the most characteristic metabolic traits of hypoxia-resistant plants.&#x0D; MATERIALS AND METHODS: Metabolite profiling was performed by GC-MS. Statistical analysis of metabolomics data was processed using R 4.3.1 Beagle Scouts.&#x0D; RESULTS: The resulting profile included 360 compounds. 74 of these were identified and 114 compounds were determined to a class. Sugars (114) were the most widely represented in the obtained profiles. 10 amino and 23 carboxylic acids, lipids and phenolic compounds have been identified. Significant differences were revealed between the profiles of leaf metabolomes of all tested species, which were clustered according to phylogenetic relation. The hydrophytic R. sceleratus, growing under submergence, showed the most unique metabolome, in which the level of sugars was reduced and intermediates of anaerobic metabolism, nitrogen metabolism, and alternative pathways of NAD(P)H reoxidation were accumulated. The profile of mesophytic R. acris was markedly different by decreased levels of amino acids, fatty acids and sterols. The metabolite profiles of waterlogged hydrophytes R. lingua and R. repens occupied an intermediate position.&#x0D; CONCLUSIONS: The identified differences of metabolomes of Ranunculus species are due to genetic determinants, ecological niche and direct impact of a stressor.

Manganese ore enhanced nitrate removal by improving bioavailability of organics released from corncob: Denitrification performance, organics transformation and nitrogen metabolic pathways

The key limiting factor in advanced nitrogen removal of carbon-poor municipal tailwaters is deficient electron donor. Corncob, a low-cost external carbon source, can enhance heterotrophic denitrification by releasing organics sustainably. But its practical application is usually restricted to low bioavailability of organics and the risk of secondary pollution. Manganese oxides (MnOx) can regulate the organics released from corncob by altering the structure and bioavailability of organics. Herein, coupling Mn Ore (MnO2 content ∼35 %) with corncob as substrate for microorganisms, quite high nitrate removal (98.4 ± 1.2 %) and less residual organics (COD < 10 mg/L) were observed. Organics characterization showed that original organics released from corncob were mainly humic substants (B/C = 0.4 ± 0.05, MW = 1.5 ∼ 350 kDa), and could be oxidized by MnOx into small molecular organics with high bioavailability (B/C = 0.72 ± 0.08, MW = 1.5 ∼ 35 kDa). Mn oxidation state analysis revealed that Mn(III/IV) in MnOx played an important role in organics transformation. The addition of corncob and redox transition of Mn enriched various functional denitrifying bacteria and dissimilatory lignocellulose -degrading bacteria, thereby upregulating functional genes encoding vital enzymes involved in nitrogen/carbon transformation. Overall, a Mn cycle-driven mixotrophic denitrification process was achieved, low molecular organics formed via MnOx oxidation accelerated heterotrophic denitrification rate, the Mn(II) produced from Mn(IV) reduction drive autotrophic denitrification. This finding could guide the low-cost technologies development for deep nitrogen removal from carbon-poor waters.

Performance of micro-pressure double-cycle coupled membrane integrated bioreactor for the treatment of urban sewage.

Based on the theory of nitrogen and phosphorus removal and technical requirements, a micro-pressure double-cycle bioreactor coupled with membrane components was used to treat municipal wastewater. The method realized the simultaneous removal of organic matter, nitrogen, and phosphorus in the same reactor and had the characteristics of membrane bioreactor process. Results showed that the average removal efficiency of COD, NH+4-N, TN, and TP were 93.74%, 95.1%, 71.85%, and 81.03%, respectively. During operation, Proteobacteria and Bacteroidetes were the main dominant bacteria, and they had complete nitrogen and phosphorus metabolic pathways. Owing to the low protein content in the mixture, the design of film placement in the micro-precipitation zone was conducive to alleviating the membrane pollution caused by the accumulation of protein, thereby improving the effluent quality and extending the service life of the membrane components.

Metagenomic insights into the mechanism for the rapid enrichment and high stability of Candidatus Brocadia facilitated by Fe(Ⅲ)

The rapid enrichment of anammox bacteria and its fragile resistance to adverse environment are the critical problems facing of anammox processes. As an abundant component in anammox bacteria, iron has been proved to promote the activity and growth of anammox bacteria in the mature anammox systems, but the functional and metabolic profiles in Fe(III) enhanced emerging anammox systems have not been evaluated. Results indicated that the relative abundance of functional genes involved in oxidative phosphorylation, nitrogen metabolism, cofactors synthesis, and extracellular polymers synthesis pathways was significantly promoted in the system added with 5 mg/L Fe(III) (R5). These enhanced pathways were crucial to energy generation, nitrogen removal, cell activity and proliferation, and microbial self-defense, thereby accelerating the enrichment of anammox bacteria Ca. Brocadia and facilitating their resistance to adverse environments. Microbial community analysis showed that the proportion of Ca. Brocadia in R5 also increased to 64.42 %. Hence, R5 could adapt rapidly to the increased nitrogen loading rate and increase the nitrogen removal rate by 108 % compared to the system without Fe(III) addition. However, the addition of 10 and 20 mg/L Fe(III) showed inhibitory effects on the growth and activity of anammox bacteria, which exhibited the lower relative abundance of Ca. Brocadia and unstable or even collapsed nitrogen removal performance. This study not only clarified the concentration range of Fe(III) that promoted and inhibited the enrichment of anammox bacteria, but also deepened our understanding of the functional and metabolic mechanisms underlying enhanced enrichment of anammox bacteria by Fe(III), providing a potential strategy to hasten the start-up of anammox from conventional activated sludge.

Towards low carbon demand and highly efficient nutrient removal: Establishing denitrifying phosphorus removal in anaerobic/anoxic/oxic + nitrification system

A two-sludge anaerobic/anoxic/oxic + nitrification system with simultaneous nitrogen and phosphorus removal was studied for enhanced low-strength wastewater treatment. After 158 days of operation, excellent NH4+-N, chemical oxygen demand (COD) and PO43−-P removal (99.0 %, 90.0 % and 92.0 %, respectively) were attained under a low carbon/nitrogen ratio of 5, resulting in effluent NH4+-N, COD and PO43−-P concentrations of 0.3, 30.0 and 0.5 mg/L, respectively. The results demonstrate that the anaerobic/anoxic/oxic sequencing batch reactor (A2-SBR) and nitrification sequencing batch reactor (N-SBR) had favorable denitrifying phosphorus removal and nitrification performance, respectively. High-throughput sequencing results indicate that the phosphate-accumulating organisms Dechloromonas (1.1 %) and Tetrasphaera (1.2 %) were enriched in the A2-SBR, while the ammonia-oxidizing bacteria Nitrosomonas (7.8 %) and the nitrite-oxidizing bacteria Nitrospira (18.1 %) showed excellent accumulation in the N-SBR. Further analysis via functional prediction revealed that denitrification is the primary pathway of nitrogen metabolism throughout the system. Overall, the system achieved low carbon and high efficiency nutrient removal.

Formation of autotrophic nitrogen removal granular sludge driven by the dual-partition airlift internal circulation: Insights from performance assessment, community succession, and metabolic mechanism

Granular sludge has been recognized as an effective method for the application and industrialization of the anammox-based process due to its good biomass retention capacity and environmental tolerance. In this study, a one-stage autotrophic nitrogen removal (ANR) dual-partition system with airlift internal circulation was implemented for 320 days. A high nitrogen removal efficiency of 84.6% was obtained, while the nitrogen removal rate reached 1.28 g-N/L/d. ANR granular sludge dominated by Nitrosomonas and Candidatus Brocadia was successfully cultivated. Results showed that activity and abundance of functional flora first increased with granulation process, but eventually declined slightly when particle size exceeded the optimal range. Total anammox activity was observed to be significantly correlated with protein content (R2 = 0.9623) and nitrogen removal performance (R2 = 0.8796). Correlation network revealed that AnAOB had complex interactions with other bacteria, both synergy for nitrogen removal and competition for substrate. Changes in abundances of genes encoding the Carbohydrate Metabolism, Energy Metabolism, and Membrane Transport suggested energy production and material transfer were possibly blocked with further sludge granulation. Formation of ANR granular sludge promoted the interactions and metabolism of functional microorganisms, and the complex nitrogen metabolic pathways improved the performance stability. These results validated the feasibility of granule formation in the airlift dual-partition system and revealed the response of the ANR system to sludge granulation.

Bioinformatic Approach to Investigate Larvae Gut Microbiota Cellulosimicrobium protaetiae via Whole-Genome Analysis.

The insect larvae Protaetia brevitarsis seulensis have recently been researched as a nutritious food source and concentrated on their environmental impacts. Therefore, their gut microbiota has been studied to elucidate their effects and roles on the environment. Of the abundance of bacterial genus identified based on the 16S rRNA genes from isolates of the gut of insect larva Protaetia brevitarsis seulensis, six of the prominent genus were identified as Bacillus (40.2%), Cellulosimicrobium (33.5%), Microbacterium (2.8%), Streptomyces (3%), Krasilnikoviella (17.5%), and Isoptericola (3%) and their similarity of 16S rRNA blast changed from 99 to 100%. Cellulosimicrobium protaetiae BI34T showed strong denitrification and cellulose degradation activity. The newly complete genome sequence of BI34T and the genomes of five species was published in the genus Cellulosimicrobium with emphasis on the denitrification and secondary metabolite genes. In order to elucidate the relationship between the strain BI34T and the host insect larva, the whole-genome sequence was analyzed and compared with the genomes of five strains in the same genus, Cellulosimicrobium, loaded from GenBank. Our results revealed the composition of the gut microbiota of the insect larvae and analyzed the genomic data for the new strain to predict its characteristics and to understand the nitrogen metabolism pathway.

Scallop-bacteria symbiosis from the deep sea reveals strong genomic coupling in the absence of cellular integration.

Previous studies have revealed tight metabolic complementarity between bivalves and their endosymbiotic chemosynthetic bacteria, but little is known about their interactions with ectosymbionts. Our analysis of the ectosymbiosis between a deep-sea scallop (Catillopecten margaritatus) and a gammaproteobacterium showed that bivalves could be highly interdependent with their ectosymbionts as well. Our microscopic observation revealed abundant sulfur-oxidizing bacteria (SOB) on the surfaces of the gill epithelial cells. Microbial 16S rRNA gene amplicon sequencing of the gill tissues showed the dominance of the SOB. An analysis of the SOB genome showed that it is substantially smaller than its free-living relatives and has lost cellular components required for free-living. Genomic and transcriptomic analyses showed that this ectosymbiont relies on rhodanese-like proteins and SOX multienzyme complex for energy generation, mainly on the Calvin-Benson-Bassham (CBB) cycle and peripherally on a phosphoenolpyruvate carboxylase for carbon assimilation. Besides, the symbiont encodes an incomplete tricarboxylic acid (TCA) cycle. Observation of the scallop's digestive gland and its nitrogen metabolism pathways indicates it does not fully rely on the ectosymbiont for nutrition. Analysis of the host's gene expression provided evidence that it could offer intermediates for the ectosymbiont to complete its TCA cycle and some amino acid synthesis pathways using exosomes, and its phagosomes, endosomes, and lysosomes might be involved in harvesting nutrients from the symbionts. Overall, our study prompts us to rethink the intimacy between the hosts and ectosymbionts in Bivalvia and the evolution of chemosymbiosis in general.