Denitrifying Bacterial Communities Research Articles

Salinity and pH related microbial nitrogen removal in the largest coastal lagoon of Chinese mainland (Pinqing Lagoon)

Coastal lagoon is critical habitat for human and provides a wide range of ecosystem services. These vital habitats are now threatened by waste discharge and eutrophication. Previous studies suggest that the pollution mitigation of coastal lagoon relies on the water exchange with open sea, and the role of microbial processes inside the lagoon is overlooked. This study takes the Pinqing Lagoon which is the largest coastal lagoon in Chinese mainland as example. The distribution of nutrients, microbial activity of nitrogen removal and community structure of denitrifying bacteria in sediment are analyzed. The results showed that the nutrient in sediment represented by DIN (1.65–12.78 mg kg−1), TOM (0.59–8.72 %) and TN (0.14–1.93 mg g−1) are at high levels and are enriched at the terrestrial impacted zone (TZ). The microbial nitrogen removal is active at 0.27–19.76 μmol N kg−1 h−1 in sediment and denitrification is the dominate pathway taking 51.44–98.71 % of total N removal. The composition of the denitrifying microbial community in marine impacted zone (MZ) is close to that of ocean and estuary, but differs considerably with those of TZ and transition zone (TM). The denitrification activity is mainly controlled by salinity and pH, and the denitrifying bacterial community composition related to the nutrient parameters of TN, TOM, etc. Our study suggested that the distribution of nutrients, microbial activity of nitrogen removal and community structure in Lagoon are the combined effects of terrestrial input and exchange with open sea. The microbial processes play important role in the nitrogen removal of coastal lagoon.

Nitrous oxide reduction by two partial denitrifying bacteria requires denitrification intermediates that cannot be respired.

Denitrification is a form of anaerobic respiration wherein nitrate (NO3-) is sequentially reduced via nitrite (NO2-), nitric oxide, and nitrous oxide (N2O) to dinitrogen gas (N2) by four reductase enzymes. Partial denitrifying bacteria possess only one or some of these four reductases and use them as independent respiratory modules. However, it is unclear if partial denitrifiers sense and respond to denitrification intermediates outside of their reductase repertoire. Here, we tested the denitrifying capabilities of two purple nonsulfur bacteria, Rhodopseudomonas palustris CGA0092 and Rhodobacter capsulatus SB1003. Each had denitrifying capabilities that matched their genome annotation; CGA0092 reduced NO2- to N2, and SB1003 reduced N2O to N2. For each bacterium, N2O reduction could be used both for electron balance during growth on electron-rich organic compounds in light and for energy transformation via respiration in darkness. However, N2O reduction required supplementation with a denitrification intermediate, including those for which there was no associated denitrification enzyme. For CGA0092, NO3- served as a stable, non-catalyzable molecule that was sufficient to activate N2O reduction. Using a β-galactosidase reporter, we found that NO3- acted, at least in part, by stimulating N2O reductase gene expression. In SB1003, NO2- but not NO3- activated N2O reduction, but NO2- was slowly removed, likely by a promiscuous enzyme activity. Our findings reveal that partial denitrifiers can still be subject to regulation by denitrification intermediates that they cannot use.IMPORTANCEDenitrification is a form of microbial respiration wherein nitrate is converted via several nitrogen oxide intermediates into harmless dinitrogen gas. Partial denitrifying bacteria, which individually have some but not all denitrifying enzymes, can achieve complete denitrification as a community by cross-feeding nitrogen oxide intermediates. However, the last intermediate, nitrous oxide (N2O), is a potent greenhouse gas that often escapes, motivating efforts to understand and improve the efficiency of denitrification. Here, we found that at least some partial denitrifying N2O reducers can sense and respond to nitrogen oxide intermediates that they cannot otherwise use. The regulatory effects of nitrogen oxides on partial denitrifiers are thus an important consideration in understanding and applying denitrifying bacterial communities to combat greenhouse gas emissions.

Homogeneous environmental selection mainly determines the denitrifying bacterial community in intensive aquaculture water.

Nitrate reduction by napA (encodes periplasmic nitrate reductase) bacteria and nitrous oxide reduction by nosZ (encodes nitrous oxide reductase) bacteria play important roles in nitrogen cycling and removal in intensive aquaculture systems. This study investigated the diversity, dynamics, drivers, and assembly mechanisms of total bacteria as well as napA and nosZ denitrifiers in intensive shrimp aquaculture ponds over a 100-day period. Alpha diversity of the total bacterial community increased significantly over time. In contrast, the alpha diversity of napA and nosZ bacteria remained relatively stable throughout the aquaculture process. The community structure changed markedly across all groups over the culture period. Total nitrogen, phosphate, total phosphorus, and silicate were identified as significant drivers of the denitrifying bacterial communities. Network analysis revealed complex co-occurrence patterns between total, napA, and nosZ bacteria which fluctuated over time. A null model approach showed that, unlike the total community dominated by stochastic factors, napA and nosZ bacteria were primarily governed by deterministic processes. The level of determinism increased with nutrient loading, suggesting the denitrifying community can be manipulated by bioaugmentation. The dominant genus Ruegeria may be a promising candidate for introducing targeted denitrifiers into aquaculture systems to improve nitrogen removal. Overall, this study provides important ecological insights into aerobic and nitrous oxide-reducing denitrifiers in intensive aquaculture, supporting strategies to optimize microbial community structure and function.

Mitigation of soil N2O emissions by decomposed straw based on changes in dissolved organic matter and denitrifying bacteria

The return of decomposed straw represents a less explored potential option for reducing N2O emissions. However, the mechanisms underlying the effects of decomposed straw return on soil N2O mitigation are still not fully clear. Therefore, we used a helium atmosphere robotized continuous flow incubation system to compare the soil N2O and N2 emissions from four treatments: CK (control: no straw), WS (wheat straw), IWS (wheat straw decomposed with Irpex lacteus), and PWS (wheat straw decomposed with Phanerochaete chrysosporium). All the treatments have been fertilized with the same amount of KNO3. Furthermore, we also analyzed i) the chemodiversity of soil dissolved organic matter (DOM), ii) the nirS, nirK, and nosZ gene copies and relative abundances of denitrifying bacterial communities (DBCs), and iii) the specific linkages between N2O emissions and DOM and DBC. The results showed that the WS, IWS and PWS treatments increased N2O emissions compared to the CK treatment. However, applying decomposed straw to soil, especially straw treated with P. chrysosporium, effectively decreased the soil N2O and increased N2 emissions compared to WS and IWS. Moreover, the IWS and PWS treatments increased the CHO composition, but they decreased the CHON and CHOS compositions of heteroatomic compounds of DOM compared with the WS and CK treatments. Furthermore, the WS, IWS and PWS treatments all significantly increased the nirS and nosZ gene copies compared with the CK treatment. Additionally, compared with the other treatments, the PWS treatment significantly shaped the DBC and led to a higher relative abundance of Pseudomonas with nirS and nosZ genes. Meanwhile, Network analysis showed that the mitigation of N2O was closely related to particular DOM molecules, and specific DBC taxa. These results highlight the potential for decomposed straw amendments to mitigate of soil N2O emissions not only by changing soil DOM but also mediating the soil DBC.

Changes of Soil Nitrogen Fractions and nirS-Type Denitrifier Microbial Community in Response to N Fertilizer in the Semi-Arid Area of Northeast China

The denitrification process is one of the important processes in the soil nitrogen (N) cycle, and is closely related to the loss of soil N fertilizer. Five treatments were included in this study: N0 (control, no N application); N90 (N application rate 90 kg ha−1); N150 (N application rate 150 kg ha−1); N210 (N application rate 210 kg ha−1); and N270 (N application rate 270 kg ha−1), to study the effect of different N application rates on the soil nirS-type denitrifying bacterial community structure, the influence of key enzyme activities during the denitrification process, and the main environmental factors affecting the variation of the denitrifying bacterial community in maize field soil under the mulched fertigation system in the semi-arid region of Northeast China. The results showed that increasing N fertilizer application significantly increased the contents of soil inorganic N and acid-hydrolyzable organic N, but significantly decreased pH. N fertilizer significantly increased nitrate reductase (NAR) activity and nitrite reductase (NIR) activity. Excessive application of N fertilizer significantly increased the nirS gene copy numbers, and, at the same time, significantly decreased the diversity of nirS-type soil denitrifying bacteria. Proteobacteria was the dominant denitrifying phylum in all treatments, and N application promoted the growth of Bradyrhizobium belonging to this phylum. The application of N fertilizer significantly changed the community structure of nirS denitrifying bacteria, and the NO3−-N content was the most important factor for this observation. Soil organic matter (SOM) and non-hydrolyzable N (NHN) indirectly affected the denitrifying bacterial community structure through NAR activity and NIR activity, while soil total N (TN) and nitrate N (NO3−-N) indirectly affected yield through denitrifying bacterial community structure. Although increasing N fertilizer was beneficial in increasing soil nutrients, the community structure of nirS-type denitrifying bacteria changed significantly. This is attributed to the increase in soil NO3−-N accumulation caused by a large amount of N application. The results of this research provide an important scientific basis for further research on the response mechanism of farmland soil denitrifying microorganisms to different N fertilizer managements under the mulched fertigation system in the semi-arid region of Northeast China.

Incorporating straw into intensively farmed cropland soil can reduce N2O emission via inhibition of nitrification and denitrification pathways

Straw incorporation (SI) combined with N fertilizer has been shown to affect soil N2O emission and N-related functional microbes in agriculture. However, the responses of N2O emission, community structure of nitrifiers and denitrifiers and related microbial functional genes to straw management strategies in the winter wheat season in China remain unclear. Here, we conducted a two-season experiment in a winter wheat field in Ningjing County, northern China, to examine four treatments: no fertilizer with (N0S1) and without maize straw (N0S0); N fertilizer with (N1S1) and without maize straw (N1S0), and their effects on N2O emissions, soil chemical parameters, crop yield, as well as the dynamics of nitrifying and denitrifying microbial communities. We found that seasonal N2O emissions decreased by 7.1–11.1% (p < 0.05) in N1S1 as compared to N1S0, without significant difference between N0S1 and N0S0. In combination with N fertilization, SI increased the yield by 2.6–4.3%, altered the microbial community composition, increased Shannon and ACE indexes, and decreased the abundance of AOA (9.2%), AOB (32.2%; p < 0.05), nirS (35.2%; p < 0.05), nirK (21.6%; p < 0.05) and nosZ (19.2%). However, in the absence of N fertilizer, SI promoted the major genera of Nitrosavbrio (AOB), unclassifiied_Gammaproteobacteria, Rhodanobacter (nirS), Sinorhizobium (nirK), which strongly correlated positively with N2O emissions. Thereby, a negative interaction effect between SI and N fertilizer on AOB and nirS emphasized that SI could offset the increase of N2O emission caused by fertilization. Soil moisture and NO3− concentration were the major factors affecting N-related microbial community structure. Our study reveals that SI suppressed N2O emission significantly and simultaneously decreased the abundance of N-related functional genes and altered denitrifying bacterial community composition. We conclude that SI helps to enhance yield and alleviate fertilizer-induced environmental costs in intensively farmed fields in northern China.

Soil glomalin-related protein affects aggregate N2O fluxes by modulating denitrifier communities in a fertilized soil

Agricultural ecosystems contribute significantly to atmospheric emissions of soil nitrous oxide (N2O), which exacerbate environmental pollution and contribute to global warming. Glomalin-related soil protein (GRSP) stabilizes soil aggregates and enhances soil carbon and nitrogen storage in agricultural ecosystems. However, the underlying mechanisms and relative importance of GRSP on N2O fluxes within soil aggregate fraction remain largely unclear. We examined the GRSP content, denitrifying bacterial community composition, and potential N2O fluxes across three aggregate-size fractions (2000–250 μm, 250–53 μm, and <53 μm) under a long-term fertilization agricultural ecosystem, subjected to mineral fertilizer or manure and their combination. Our findings indicated that various fertilization treatments have no discernible impact on the size distribution of soil aggregates, paving the way to further research into the impact of soil aggregates on GRSP content, the denitrifying bacterial community composition, and potential N2O fluxes. GRSP content increased with the increase in soil aggregate size. Potential N2O fluxes (including gross N2O production and N2O reduction and net N2O production) among aggregates were highest in microaggregates (250–53 μm), followed by macroaggregates (2000–250 μm) and lowest in silt + clay (<53 μm) fractions. Potential N2O fluxes had a positive response to soil aggregate GRSP fractions. The non-metric multidimensional scaling analysis revealed that soil aggregate size could drive the denitrifying functional microbial community composition, and deterministic processes play more critical roles than stochasticity processes in driving denitrifying functional composition under soil aggregate fractions. Procrustes analysis revealed a significant correlation between denitrifying microbial community, soil aggregate GRSP fractions, and potential N2O fluxes. Our study suggests that soil aggregate GRSP fractions influence potential nitrous oxide fluxes by affecting denitrifying microbial functional composition within soil aggregate.

Novel insights into aerobic denitrifying bacterial communities augmented denitrification capacity and mechanisms in lake waters

Scanty attention has been paid to augmenting the denitrification performance of polluted lake water by adding mix-cultured aerobic denitrifying bacterial communities (Mix-CADBCs). In this study, to solve the serious problem of nitrogen pollution in lake water bodies, aerobic denitrifying bacteria were added to lake water to enhance the nitrogen and carbon removal ability. Three Mix-CADBCs were isolated from lake water and they could remove >94 % of total nitrogen and dissolved organic carbon, respectively. The balance of nitrogen analysis shown that >70 % of the initial nitrogen was converted to gaseous nitrogen, and <11 % of the initial nitrogen was converted into microbial biomass. The batch experiments indicated that three Mix-CADBCs could perform denitrification under various conditions. According to the results of nirS-type sequencing, the Hydrogenophaga sp., Prosthecomicrobium sp., and Pseudomonas sp. were dominated genera of three Mix-CADBCs. The analysis of network indicated Pseudomonas I.Bh25.14 and Vogsella LIG4 were correlated with the removal of total nitrogen (TN) and dissolved organic carbon in the Mix-CADBCs. Compared with lake raw water, the addition of three Mix-CADBCs could promote the denitrification capacity (the removal efficiencies of TN > 78.72 %), microbial growth (optical density increased by 0.015–0.138 and the total cell count increased by 2 times), and organic degradation ability (the removal efficiency chemical oxygen demand >38 %) of lake water. In general, the findings of this study demonstrated that Mix-CADBCs could provide a new perspective for biological treatment lake water body.

Presence of algal symbionts affects denitrifying bacterial communities in the sea anemone Aiptasia coral model

The coral-algal symbiosis is maintained by a constant and limited nitrogen availability in the holobiont. Denitrifiers, i.e., prokaryotes reducing nitrate/nitrite to dinitrogen, could contribute to maintaining the nitrogen limitation in the coral holobiont, however the effect of host and algal identity on their community is still unknown. Using the coral model Aiptasia, we quantified and characterized the denitrifier community in a full-factorial design combining two hosts (CC7 and H2) and two strains of algal symbionts of the family Symbiodiniaceae (SSA01 and SSB01). Strikingly, relative abundance of denitrifiers increased by up to 22-fold in photosymbiotic Aiptasia compared to their aposymbiotic (i.e., algal-depleted) counterparts. In line with this, while the denitrifier community in aposymbiotic Aiptasia was largely dominated by diet-associated Halomonas, we observed an increasing relative abundance of an unclassified bacterium in photosymbiotic CC7, and Ketobacter in photosymbiotic H2, respectively. Pronounced changes in denitrifier communities of Aiptasia with Symbiodinium linucheae strain SSA01 aligned with the higher photosynthetic carbon availability of these holobionts compared to Aiptasia with Breviolum minutum strain SSB01. Our results reveal that the presence of algal symbionts increases abundance and alters community structure of denitrifiers in Aiptasia. Thereby, patterns in denitrifier community likely reflect the nutritional status of aposymbiotic vs. symbiotic holobionts. Such a passive regulation of denitrifiers may contribute to maintaining the nitrogen limitation required for the functioning of the cnidarian-algal symbiosis.

Effects of Nitrogen Input on Community Structure of the Denitrifying Bacteria with Nitrous Oxide Reductase Gene (nosZ I): a Long-Term Pond Experiment.

Excessive nitrogen (N) input is an important factor influencing aquatic ecosystems and has received increasing public attention in the past decades. It remains unclear how N input affects the denitrifying bacterial communities that play a key role in regulating N cycles in various ecosystems. To test our hypothesis-that the abundance and biodiversity of denitrifying bacterial communities decrease with increasing N-we compared the abundance and composition of denitrifying bacteria having nitrous oxide reductase gene (nosZ I) from sediments (0-20 cm) in five experimental ponds with different nitrogen fertilization treatment (TN10, TN20, TN30, TN40, TN50) using quantitative PCR and pyrosequencing techniques. We found that (1) N addition significantly decreased nosZ I gene abundance, (2) the Invsimpson and Shannon indices (reflecting biodiversity) first increased significantly along with the increasing N loading in TN10-TN40 followed by a decrease in TN50, (3) the beta diversity of the nosZ I denitrifier was clustered into three groups along the TN concentration levels: Cluster I (TN50), Cluster II (TN40), and Cluster III (TN10-TN30), (4) the proportions of Alphaproteobacteria and Betaproteobacteria in the high-N treatment (TN50) were significantly lower than in the lower N treatments (TN10-TN30). (5) The TN concentration was the most important factor driving the alteration of denitrifying bacteria assemblages. Our findings shed new light on the response of denitrification-related bacteria to long-term N loading at pond scale and on the response of denitrifying microorganisms to N pollution.

Aerobic denitrifying bacteria community structure and response of aerobic denitrifying bacteria to dissloved organic matter in the sediments in Lake Baiyangdian in the spring

好氧反硝化因其独特优势成为近年来生物脱氮的研究热点，溶解性有机物（DOM）作为微生物碳源是造成群落差异的重要原因，为了探究白洋淀不同功能区好氧反硝化菌群落结构对溶解性有机物碳源的响应，本文结合荧光区域积分法以及napA反硝化基因的高通量测序技术，对白洋淀春季沉积物中的好氧反硝化菌群落结构特征以及好氧反硝化菌对溶解性有机物的响应进行研究.结果表明，春季白洋淀沉积物有机质组分类蛋白质组分高于类腐殖质组分，其中养殖区的类蛋白质组分最大，达到79.63%±3.79%，原始区的类腐殖质组分最大，达到33.91%±6.32%；高通量测序得到3693个OTUs，共分为9个主要门类，其中，变形菌门占比最大，达到99%以上；α多样性Chao1指数呈现显著的空间差异，并且旅游区>生活区>入淀区>原始区>养殖区；该时期好氧反硝化菌主要物种组成包括Aeromonas、Sulfuritortus、Cupriavidus、Pseudomonas和Thauera，Ferrimonas作为指示物种，差异贡献最大；冗余分析发现，可见光区类富里酸物质和微生物代谢产物组分分别是不同功能区好氧反硝化菌群落纲水平和属水平差异的主要原因.综上，通过对白洋淀好氧反硝化菌群与溶解性有机物的相关关系研究，不仅有助于进一步认识天然环境中氮循环微生物的特征；还可以为将来适于实际环境的高效菌筛选的碳源选择提供参考.;Aerobic denitrification has become a research hotspot in biological denitrification in recent years due to its unique advantages. Dissolved organic matter (DOM) as a carbon source is the important reason for community differences. To explore the responding mechanism of aerobic denitrifying bacteria community structure to dissolved organic matter in different functional areas of Lake Baiyangdian, this paper combines the fluorescence area integration method and the high-throughput sequencing technology of napA denitrification genes to study the characteristics of the aerobic denitrifying bacterial community structure and responding mechanism in Lake Baiyangdian. The results showed that the concentration of protein-like components was higher than that of humic-like components in the organic matter of Lake Baiyangdian sediments in spring. Among them, the protein-like components in the culture area were the largest, reaching 79.63%±3.79%, and the humus-like component in the primitive area was the largest, reaching 33.91%±6.32%; A total of 3693 OTUs were acquired high Throughput sequencing, which was divided into 9 main phyla. Among them, the Proteobacteria accounted for the largest proportion, reaching over 99%; the α diversity Chao1 index showed significant spatial differences, and the travel area>the living area>the entry area>the original area>the breeding area; the main species of aerobic denitrifying bacteria include Aeromonas, Sulfuritortus, Cupriavidus, Pseudomonas and Thaurea, Ferrimonas, as an indicator species, contributes the most to the difference; Redundant analysis found that the components of fulvic acid-like substances and microbial metabolites in the visible light area are the main reasons for the differences in aerobic denitrifying bacteria in different functional areas. In summary, this research will not only help to further understand the characteristics of the nitrogen cycle microorganisms in the natural environment; it will also provide a reference for the selection of carbon sources suitable for efficient bacterial screening in the practical environment.

Effects of monoculture regime on the soil nirK- and nosZ-denitrifying bacterial communities of Casuarina equisetifolia

The forest quality of Casuarina equisetifolia has reduced dramatically due to soil sickness under continuous monoculture. The effect of microbes on the growth of C. equisetifolia plants has been extensively investigated. In this study, quantitative PCR assays and Illumina MiSeq sequencing of targeted nirK and nosZ genes were carried out to identify the variations in abundance in the soil and community structure of denitrifying bacteria under the long-term consecutive monoculture of C. equisetifolia. The principal component analysis illustrated that the denitrifying bacterial community structure was obviously different among the soils in the treatments of no C. equisetifolia cultivation (CK), the first continuous plantation (FCP), the second continuous plantation (SCP), and the third continuous plantation (TCP). Taxonomic analysis demonstrated that in both the nirK and nosZ gene-containing denitrifying bacterial communities, Proteobacteria increased significantly in C. equisetifolia monocultures. Furthermore, the genera Bosea, Sinorhizobium, Streptomyces, Azospirillum, Mesorhizobium, Thiobacillus, Ochrobactrum, and Rhodoferax were enriched, while Rhodopseudomonas, Pseudomonas, Chelativorans, and Chelatococcus decreased under continuous monoculture (SCP and TCP). Redundancy analysis showed that Streptomyces, Pseudomonas, and Rhodopseudomonas were positively correlated with soil physicochemical factors. In summary, C. equisetifolia monoculture led to dramatic shifts in the quantity and community structure of denitrifying bacteria, contributing greatly to soil sickness in this tree species.

Mix-cultured aerobic denitrifying bacterial communities reduce nitrate: Novel insights in micro-polluted water treatment at lower temperature

Three mix-cultured aerobic denitrifiers were screened from a source water reservoir and named HE1, HE3 and SU4. Approximately 72.9%, 68.6% and 66.2% of nitrate were effectively removed from basal medium, respectively, after 120 h of cultivation at 8 °C. The nitrogen balance analysis revealed about one-fifth of the initial nitrogen was converted into gaseous denitrification products. According to the results of Biolog, the three microfloras had high metabolic capacity to carbon sources. The dominant genera were Pseudomonas and Paracoccus in these bacterial communities based on nirS gene sequencing. Response surface methodology elucidated that the denitrification rates of identified bacteria reached the maximum under the following optimal parameters: C/N ratio of 7.51–8.34, pH of 8.03–8.09, temperature of 18.03–20.19 °C, and shaking speed of 67.04–120 rpm. All results suggested that screened aerobic denitrifiers could potentially be applied to improve the source water quality at low temperature.

Effects of truffle inoculation on a nursery culture substrate environment and seedling of Carya illinoinensis

We inoculated Tuber aestivum and Tuber sinoaestivum on Carya illinoinensis to explore the effects of inoculation on host plant growth, enzyme activities, the physicochemical properties of rhizosphere soil, the denitrifying bacterial community in the rhizosphere, and the distribution of mating type genes in the rhizosphere. We found that the Tuber spp. inoculation increased the height of the host plant and that the stem circumference of the host was greater two months after inoculation. Six months after inoculation, the peroxidase activity of the seedlings inoculated with T. sinoaestivum was higher than that of the control. At four and six months after inoculation, the superoxidase dismutase activities of the seedlings inoculated with T. aestivum were higher than those of the seedlings inoculated with T. sinoaestivum. Six months after inoculation, nitrate nitrogen content was lowest in the control and highest in the T. sinoaestivum treatment. Among the nirS-type denitrifying bacteria community, the relative abundances of Proteobacteria were high. T. aestivum and T. sinoaestivum inoculation did not affect the diversity of denitrifying bacteria. The mating type genes MAT1-1-1 and MAT1-2-1 were detected in the rhizosphere of C. illinoinensis inoculated with T. sinoaestivum and T. aestivum, and MAT1-1-1 dominated over MAT1-2-1.

Total and denitrifying bacterial communities associated with the interception of nitrate leaching by carbon amendment in the subsoil.

Nitrate leaching is severe in greenhouse where excessive nitrogen is often applied to maintain high crop productivities. In this study, we investigated the effects of carbon amendment in the subsoil on nitrate leaching and the emission of greenhouse gases (CH4 and N2O) using a soil column experiment. Carbon amendment resulted in over 39% reduction in nitrate leaching and 25.3% to 60.6% increase of total N content in the subsoil zone as compared to non-amended control. Strikingly, the abundance of nirS, nosZ, and 16S rRNA were higher in the treatment than the corresponding controls while no significant effect was detected for nirK. Carbon amendment explained 14%, 10%, and 4% of the variation in the community of nosZ, nirS, and nirK, respectively. It also considerably (more than 7 times) enriched genera such as Anaerovorax, Pseudobacteroides, Magnetospirillum, Prolixibacter, Sporobacter, Ignavibacterium, Syntrophobacter, Oxobacter, Hydrogenispora, Desulfosporomusa, Mangrovibacterium, and Sporomusa, as revealed by the analysis of 16S rRNA amplicon. Network analysis further uncovered that carbon amendment enriched three microbial hubs which mainly consists of positively correlated nirS, nosZ, and anaerobic bacterial populations. In summary, carbon amendment in the subsoil mitigated nitrate leaching and increased the nitrogen pool by possible activation of denitrifying and anaerobic bacterial populations. KEY POINTS: • Carbon amendment in subsoil reduced NO3- leaching by over 39% under high N input. • Carbon amendment increased the total N in subsoil from 25.3% to 60.6%. • Carbon amendment enriched nirS- and nosZ-type denitrifying bacteria in subsoil.

Long-term effects of decabromodiphenyl ether on denitrification in eutrophic lake sediments: Different sensitivity of six-type denitrifying bacteria

The widespread use of polybrominated diphenyl ethers inevitably results in their increased release into natural waters and subsequent deposition in sediments. However, their long-term effects on the bacteria participating in each step of denitrification in eutrophic lake sediments are still unknown. Here, we conducted a one-year microcosm experiment to determine the long-term effects of decabromodiphenyl ether (BDE-209), at low (2 mg kg−1 dry weight) and high (20 mg kg−1 dry weight) contamination levels, on six-type denitrifying bacteria and their activities in sediments collected from Taihu Lake, a typical eutrophic lake in China. At the end of the experiment, sediment denitrifying reductase activities were inhibited by BDE-209 at both levels, with the greatest inhibition seen for nitric oxide reductase activity. The higher nitrate concentration in the contaminated sediments was attributed to the inhibition of nitrate reductase activities. The abundances of six-type denitrifying genes (narG, napA, nirK, nirS, norB, and nosZ) significantly decreased under high BDE-209 treatment, and narG and napA genes were more sensitive to the toxicity of BDE-209. The results from pyrosequencing showed that BDE-209, at either treatment concentration, decreased the six-type denitrifying bacterial diversities and altered their community composition. This shift of six-type denitrifying bacterial communities might also be driven by the debrominated products concentrations of BDE-209 and variations in sediment inorganic nitrogen concentrations. In particular, some genera from phylum Proteobacteria such as Pseudomonas, Cupriavidus, and Azoarcus were decreased significantly because of BDE-209 and its debrominated products.

Emerging investigator series: prompt response of estuarine denitrifying bacterial communities to copper nanoparticles at relevant environmental concentrations

In estuaries the deposition of copper nanoparticles upon sediments can contribute to change metal availability and promote the transcriptional response of denitrifying bacteria.

Aerobic denitrifying bacterial communities drive nitrate removal: Performance, metabolic activity, dynamics and interactions of core species

Three novel mix-cultured aerobic denitrifying bacteria (Mix-CADB) consortia named D14, X21, and CL exhibited excellent total organic carbon (TOC) removal and aerobic denitrification capacities. The TOC and nitrate removal efficiencies were higher than 93.00% and 98.00%. The results of Biolog demonstrated that three communities displayed high carbon metabolic activity. nirS gene sequencing and ecological network model revealed that Pseudomonas stutzeri, Paracoccus sp., and Paracoccus denitrificans dominated in the D14, X21, and CL communities. The dynamics and co-existence of core species in communities drove the nutrient removal. Response surface methodology showed the predicted total nitrogen removal efficiency reached 99.43% for D14 community. The three Mix-CADB consortia have great potential for nitrogen-polluted aquatic water treatment because of their strong adaptability and removal performance. These results will provide new understanding of co-existence, interaction and dynamics of Mix-CADB consortia for nitrogen removal in nitrogen-polluted aquatic ecosystems.

Effects of inoculation with lignocellulose-degrading microorganisms on nitrogen conversion and denitrifying bacterial community during aerobic composting

The present study compared the effects of inoculation (WSD treatment) and non-inoculation (CK treatment) with lignocellulose-degrading microorganisms on nitrogen conversion, nitrogen functional genes, and the denitrifying bacterial community during aerobic composting, and their potential relations to NH3 and N2O emissions were also explored. Results showed that, WSD reduced the NH3 and N2O emissions by 25.9% and 34.98%, respectively, compared with CK. WSD also reduced the abundances of nitrifying (bacteria amoA) and denitrifying (nirS, nirK, and nosZ) genes during composting, which were significantly positively correlated with N2O emissions (P < 0.01). The most important nosZ denitrifying microorganisms belonged to Proteobacteria. Redundancy analysis showed that environmental factors could affect the succession of the denitrifying bacterial community during composting. Based on these results, structural equation modeling demonstrated that the reduction in N2O emissions under WSD was related to the lower accumulation of NO3–-N utilized by denitrifying microorganisms during the compost maturation period.

Nitrifying and Denitrifying Microbial Communities in Centralized and Decentralized Biological Nitrogen Removing Wastewater Treatment Systems

Biological nitrogen removal (BNR) in centralized and decentralized wastewater treatment systems is assumed to be driven by the same microbial processes and to have communities with a similar composition and structure. There is, however, little information to support these assumptions, which may impact the effectiveness of decentralized systems. We used high-throughput sequencing to compare the structure and composition of the nitrifying and denitrifying bacterial communities of nine onsite wastewater treatment systems (OWTS) and one wastewater treatment plant (WTP) by targeting the genes coding for ammonia monooxygenase (amoA) and nitrous oxide reductase (nosZ). The amoA diversity was similar between the WTP and OWTS, but nosZ diversity was generally higher for the WTP. Beta diversity analyses showed the WTP and OWTS promoted distinct amoA and nosZ communities, although there is a core group of N-transforming bacteria common across scales of BNR treatment. Our results suggest that advanced N-removal OWTS have microbial communities that are sufficiently distinct from those of WTP with BNR, which may warrant different management approaches.