Enhanced nitrogen removal of the anaerobic ammonia oxidation process by coupling with an efficient nitrate reducing bacterium (Bacillus velezensis M3-1).

Bacillus velezensis M3-1 strain isolated from the sediment of Myriophyllum aquatium constructed wetlands was found to efficiently convert NO3--N to NO2--N, and the requirements for carbon source addition were not very rigorous. This work demonstrates, for the first time, the feasibility of using the synergy of anammox and Bacillus velezensis M3-1 microorganisms for nitrogen removal. In this study, the possibility of M3-1 that converted NO3--N produced by anammox to NO2--N was verified in an anaerobic reactor. The NO3--N reduction ability of M3-1 and denitrifying bacteria in coupling system was investigated under different C/N conditions, and it was found that M3-1 used carbon sources preferentially over denitrifying bacteria. By adjusting the ratio of NH4+-N to NO2--N, it was found that the NO2--N converted from NO3--N by M3-1 participated in the original anammox.The nitrogen removal efficacy (NRE) of the coupled system was increased by 12.1%, compared to the control group anammox system at C/N = 2:1. Functional gene indicated that it might be a nitrate reducing bacterium.This study shows that the nitrate reduction rate achieved by the Bacillus velezensis M3-1 can be high enough for removing nitrate produced by anammox process, which would enable improve nitrogen removal from wastewater.

Precipitation Drives Soil Protist Diversity and Community Structure in Dry Grasslands.

Protists are essential components of soil microbial communities, mediating nutrient cycling and ecosystem functions in terrestrial ecosystems. However, their distribution patterns and driving factors, particularly, the relative importance of climate, plant and soil factors, remain largely unknown. This limits our understanding of soil protist roles in ecosystem functions and their responses to climate change. This is particularly a concern in dryland ecosystems where soil microbiomes are more important for ecosystem functions because plant diversity and growth are heavily constrained by environmental stresses. Here, we explored protist diversity and their driving factors in grassland soils on the Tibetan Plateau, which is a typical dryland region with yearly low temperatures. Soil protist diversity significantly decreased along the gradient of meadow, steppe, and desert. Soil protist diversity positively correlated with precipitation, plant biomass and soil nutrients, but these correlations were changed by grazing. Structural equation and random forest models demonstrated that precipitation dominated soil protist diversity directly and indirectly by influencing plant and soil factors. Soil protist community structure gradually shifted along meadow, steppe and desert, and was driven more by precipitation than by plant and soil factors. Soil protist community compositions were dominated by Cercozoa, Ciliophora and Chlorophyta. In particular, Ciliophora increased but Chlorophyta decreased in relative abundance along the gradient of meadow, steppe and desert. These results demonstrate that precipitation plays more important roles in driving soil protist diversity and community structure than plant and soil factors, suggesting that future precipitation change profoundly alters soil protist community and functions in dry grasslands.

Production of biochar from squeezed liquid of fruit and vegetable waste: Impacts on soil N2O emission and microbial community.

The squeezed liquid from fruit and vegetable waste (LW) presents a unique wastewater challenge, marked by recalcitrance in treatment and amplified design risks with the application of conventional processes. Following coagulation of the squeezed liquid, the majority of particulate matter precipitates. The resulting precipitated floc (LWF) is reclaimed and subsequently utilized for the synthesis of biochar. The present study primarily explores the viability of repurposing LWF as biochar to enhance soil quality and mitigate N2O emissions. Findings indicate that the introduction of a 2% proportion of LWFB led to a remarkable 99.5% reduction in total N2O emissions in contrast to LWF. Concurrently, LWFB substantially enhanced nutrients content by elevating soil organic carbon (SOC) and nitrogen levels. Utilizing high-throughput sequencing in conjunction with qPCR, the investigation unveiled that the porous structure and substantial specific surface area of LWFB potentially fostered microbial adhesion and heightened diversity within the soil microbial community. Furthermore, LWFB notably diminished the relative abundance of AOB (Nitrosospira, Nitrosomonas), and NOB (Candidatus_Nitrotoga), thereby curbing the conversion of NH4+ into NO3-. The pronounced elevation in nosZ abundance implies that LWFB holds the potential to mitigate N2O emissions through a conversion to N2.

Transcriptomics Insights into Phosphorus Stress Response of Myriophyllum aquaticum.

Through excellent absorption and transformation, the macrophyte Myriophyllum (M.) aquaticum can considerably remove phosphorus from wastewater. The results of changes in growth rate, chlorophyll content, and roots number and length showed that M. aquaticum could cope better with high phosphorus stress compared with low phosphorus stress. Transcriptome and differentially expressed genes (DEGs) analyses revealed that, when exposed to phosphorus stresses at various concentrations, the roots were more active than the leaves, with more DEGs regulated. M. aquaticum also showed different gene expression and pathway regulatory patterns when exposed to low phosphorus and high phosphorus stresses. M. aquaticum's capacity to cope with phosphorus stress was maybe due to its improved ability to regulate metabolic pathways such as photosynthesis, oxidative stress reduction, phosphorus metabolism, signal transduction, secondary metabolites biosynthesis, and energy metabolism. In general, M. aquaticum has a complex and interconnected regulatory network that deals efficiently with phosphorus stress to varying degrees. This is the first time that the mechanisms of M. aquaticum in sustaining phosphorus stress have been fully examined at the transcriptome level using high-throughput sequencing analysis, which may indicate the direction of follow-up research and have some guiding value for its future applications.

Strategies for energy conversion from sludge to methane through pretreatment coupled anaerobic digestion: Potential energy loss or gain.

Anaerobic digestion (AD) of wasted activated sludge from wastewater plants is recognized as an effective method to reclaim energy in the form of methane. AD performance has been enhanced by coupling various pretreatments that impact energy conversion from sludge. This paper mainly reviewed the development of pretreatments based on different technologies reported in recent years and evaluated their energy benefit. Significant increases in methane yield are generally obtained in AD with pretreatments demanding energy input, including thermal- and ultrasound-based methods. However, these energy-intense pretreatments usually gained negative energy benefit that the increase in methane yield consumed extra energy input. The unbalanced relationship counts against the goal of energy reclamation from sludge. Combined pretreatment consisting of multiple technologies normally outcompetes the single pretreatment, and the combination of energy-intense methods and chemicals potentially reduces energy input and simultaneously ensure high methane yield. For determining whether the energy reclamation from sludge via AD contribute to mitigating global warming, integrating greenhouse gas emission into the evaluation system of pretreated AD is further warranted.

Purification performance from bypass ecological treatment systems treating WWTPs effluents and improvement of water quality in receiving rivers: A case study in southern China.

Effluents from wastewater treatment plants (WWTPs) is the main source of pollution in rivers in developing countries. In this case study, three bypass ecological treatment systems along urban rivers achieved high removal efficiencies for chemical oxygen demand (COD; 55.7-64.0%), ammonium N (NH4+-N; 63.1-89.4%) and total phosphorous (TP; 27.6-76.7%). 16 S rRNA gene sequencing analysis confirmed that Proteobacteria was the main bacterial phylum (44.4%) in the ecological treatment system, and members were enriched significantly in the non-aeration area (59.3%). The relative abundance of Nitrospirae was highest in the inflow area (25.0%), but restrained in the non-aeration area (5.7%). 18 S rRNA gene annotation results indicated that phylum Rotifer was gradually inhibited with the direction of water flow and diffusion, while phylum Rhodophyta displayed the opposite trend. After implementation of bypass ecological treatment systems, receiving rivers were improved significantly from Grade Ⅴ to Ⅳ, and the biodiversity of zooplankton, zoobenthos and fish communities was greatly improved.

Fast start-up of anammox process: Effects of extracellular polymeric substances addition on performance, granule properties, and bacterial community structure.

The slow startup is the major obstacle to the application of anaerobic ammonium oxidation (anammox) process in mainstream wastewater treatment. Extracellular polymeric substances (EPS) are one potential resource for stable anammox reactor operation. Response surface analysis was used to optimize the specific anammox activity (SAA) with the addition of EPS; SAA was maximum at a temperature of 35 °C and the EPS concentration of 4 mg/L. By comparing the nitrogen removal of anammox reactors with no EPS (R0), immobilized EPS (EPS-alginate beads) (R1), and liquid EPS (R2), we found that EPS-alginate beads significantly speed up the startup of anammox process and enable the start time to be shortened from 31 to 19 days. As a result of the higher MLVSS content, higher zeta potential, and lower SVI30, anammox granules of R1 exhibited a stronger capacity to aggregate. Moreover, EPS extracted from R1 had higher flocculation efficiencies than EPS derived from R0 and R2. Phylogenetic analysis of 16S rRNA genes revealed that the main anammox species in R1 is Kuenenia taxon. To clarify the relative significance of stochastic vs deterministic processes in the anammox community, neutral model and network analysis are employed. In R1, community assembly became more deterministic and stable than in other cultures. Our results show that EPS might inhibit heterotrophic denitrification and thereby promote anammox activity. This study suggested a quick start-up strategy for the anammox process based on resource recovery, which is helpful for environmentally sustainable and energy-efficient wastewater treatment.

Microbial community structure analyses and cultivable denitrifier isolation of Myriophyllum aquaticum constructed wetland under low C/N ratio.

With the rapid expansion of livestock production, the amount of livestock wastewater accumulated rapidly. Lack of biodegradable organic matter makes denitrification of livestock wastewater after anaerobic digestion more difficult. In this study, Myriophyllum aquaticum constructed wetlands (CWs) with efficient nitrogen removal performance were established under different carbon/nitrogen (C/N) ratios. Analysis of community composition reveals the change of M. aquaticum CWs in microbial community structure with C/N ratios. The proportion of Proteobacteria which is one of the dominant phyla among denitrifier communities increased significantly under low C/N ratio conditions. Besides, to obtain cultivable denitrifier that could be added into CWs in situ, 33 strains belonging to phylum Proteobacteria were isolated from efficient M. aquaticum CWs, while the best-performing denitrification strain M3-1 was identified as Bacillus velezensis JT3-1 (GenBank No. CP032506.1). Redundancy analysis and quadratic models showed that C/N ratio had significant effects on disposal of nitrate (NO3--N) and the strains isolated could perform well in denitrification when C/N ratio is relatively low. In addition, they have relatively wide ranges of carbon sources, temperature and a high NO3- removal rate of 9.12 mg/(L·hr) at elevated concentrations of 800 mg/L nitrate. Thus, strains isolated from M. aquaticum CWs with low C/N ratio have a practical application value in the treatment of nitrate-containing wastewater. These denitrifying bacteria could be added to CWs to enhance nitrogen removal efficiency of CWs for livestock wastewater with low C/N ratio in the future.

Impact of influent strengths on nitrous oxide emission and its molecular mechanism in constructed wetlands treating swine wastewater.

Constructed wetlands (CWs) can remove nitrogen (N) through plant assimilation and microbial nitrification and denitrification, while it also releases large greenhouse gas nitrous oxide (N2O) into the atmosphere. However, N2O emissions and the underlying microbial mechanisms of CWs when treating high-strength wastewater have not been systematically surveyed. Here, the effect of three influent strengths on N2O emissions in a pilot-scale CW treatment of swine wastewater was determined and the underlying microbial mechanisms were explored. The results showed that the removal rates of ammonium (NH4+) and total nitrogen (TN) increased significantly with the increasing influent strengths, however, the ratio of N2O emission/TN removal rose by 1.5 times at the same time. Quantitation of microorganisms responsible for N-cycle in the sediment indicated that the abundance of ammonia-oxidizing bacteria (AOB) in high influent strengths (COD, 962.38 ± 3.05 mg/L; NH4+, 317.89 ± 4.24 mg/L) was 51.6-fold compared with that in low influent strengths (COD, 516.94 ± 4.18 mg/L; NH4+, 100.65 ± 2.65), and AOB gradually replaced ammonia-oxidizing archaea (AOA) to dominate ammonia oxidizers. Structural equation models demonstrated that NO2- accumulations promoted the ratio of AOB/AOA, which further led to an increase in the ratio of N2O emission/TN removal. It is worth noting both the N removal rates and N2O emissions increased with the increasing influent strength. To obtain reduced N2O emissions, pretreatment technology for strength reduction should be supplemented before high-strength wastewater enters the CWs. This study may shed new light on the sustainable operation and application of CWs.

Oxidative stress of Microcystis aeruginosa induced by algicidal bacterium Stenotrophomonas sp. KT48.

Cyanobacterial harmful algal blooms are a worldwide problem with substantial adverse effects on the aquatic environment as well as human health. Among the multiple physicochemical and biotic approaches, algicidal bacterium is one of the most promising and eco-friendly ways to control bloom expansion. In this study, Stenotrophomonas sp. KT48 isolated from the pond where cyanobacterial blooms occurred exhibited a strong inhibitory effect on Microcystis aeruginosa. However, the algicidal performance and mechanisms of Stenotrophomonas sp. remain under-documented. To explore the algicidal performance and physiological response againt M. aeruginosa, further works were implemented here. Our results indicated that the algicidal rate of strain KT48 cultured in 1/8 LB medium supplemented with 0.3% starch or glucose was about 30% higher than that in 1/8 LB medium. Strain KT48 culture, cell-free filtrate, and cells re-suspended were inoculated into the M. aeruginosa culture, and the Chl-a content was determined. Those results indicated that the algicidal activity of cells re-suspended was far higher than that of cell-free filtrate and culture. Thus, strain KT48 exhibited algicidal activity mainly through direct attacking M. aeruginosa rather than excretion of algicides. Furthermore, strain KT48 led to an increase in cellular reactive oxygen species (ROS) and caused lipid peroxidation as supported by the increase in malondialdehyde (MDA) levels. The ROS and MDA levels in algal cells treated with strain KT48 cells re-suspended were about 3.23-fold and 2.80-fold higher than those of untreated algal cells on day 11. And a further inhibition to the antioxidant system is suggested by a sharp decrease in the superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities. In addition, we also observed that the morphology of most algal cells changed from integrity to break. This study not only indicated strain KT48 with strong algicidal activity, but also explored the underlying algicidal mechanisms to provide a source of bacterial agent for the biocontrol of cyanobacterial blooms. KEY POINTS: • Strain KT48 exhibited strong algicidal activity mainly through direct attacking M. aeruginosa. • The addition of glucose could enhance the algicidal rate of strain KT48 by about 30%. • Strain KT48 led to an increase in cellular reactive oxygen species (ROS) level that causes membrane damage as supported by the increase in malondialdehyde (MDA) levels.

Ecological selection of bacterial taxa with larger genome sizes in response to polycyclic aromatic hydrocarbons stress.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous priority pollutants that cause great damage to the natural environment and health. Average genome size in a community is critical for shedding light on microbiome's functional response to pollution stress within an environment. Here, microcosms under different concentrations were performed to evaluate the selection of PAHs stress on the average genome size in a community. We found the distinct communities of significantly larger genome size with the increase of PAHs concentration gradients in soils, and consistent trends were discovered in soils at different latitudes. The abundance of Proteobacteria and Deinococcus-Thermus with relatively larger genomes increased along with PAHs stress and well adapted to polluted environments. In contrast, the abundance of Patescibacteria with a highly streamlined and smaller genome decreased, implying complex interactions between environmental selection and functional fitness resulted in bacteria with larger genomes becoming more abundant. Moreover, we confirmed the increased capacity for horizontal transfer of degrading genes between communities by showing an increased connection number per node positively related to the nidA gene along the concentration gradients in the co-occurrence network. Our findings suggest PAHs tend to select bacterial taxa with larger genome sizes, with significant consequences for community stability and potential biodegradation strategies.

Efficiency and mechanism of reducing ammonia volatilization in alkaline farmland soil using Bacillus amyloliquefaciens biofertilizer.

Ammonia volatilization from the farmland caused by the application of synthetic nitrogen fertilizer is the most important source of anthropogenic ammonia emissions. Biofertilizer application has been considered as an alternative option for agriculture sustainability and soil improvement. In this study, field trials were carried out to investigate the efficiency of Bacillus amyloliquefaciens (BA) biofertilizer on alleviating ammonia volatilization in alkaline farmland soil and increasing crop yield and nitrogen utilization. Potential response mechanisms were investigated from soil enzyme, nitrogen cycle function genes and microbial community levels. Compared with conventional fertilization, BA biofertilizer application reduced the ammonia volatilization by 68%, increased the crop yield and nitrogen recovery by 19% and 19%, respectively. Soil enzyme activity analysis showed that BA biofertilizer inhibited the urease activity and enhanced the potential ammonia oxidation (PAO). In addition, BA biofertilizer application also increased the bacterial amoA gene abundance, while decreased the ureC gene abundance. BA biofertilizer also significantly altered the community structure and composition, and especially raised the abundance of ammonia oxidation bacteria (AOB), while no changes were observed in abundance of nitrite oxidation bacteria (NOB). Briefly, BA biofertilizer was approved to reduce the transformation of fertilizer nitrogen to NH4+-N, simultaneously accelerating NH4+-N into the nitrification process, thus decreasing the NH4+-N content remained in alkaline soil and consequently alleviating the ammonia volatilization. Thus, these results suggested that the application of BA biofertilizer is a feasible strategy to improve crop yields and reduce agricultural ammonia emissions.

Anaerobic degradation of xenobiotic organic contaminants (XOCs): The role of electron flow and potential enhancing strategies.

In groundwater, deep soil layer, sediment, the widespread of xenobiotic organic contaminants (XOCs) have been leading to the concern of human health and eco-environment safety, which calls for a better understanding on the fate and remediation of XOCs in anoxic matrices. In the absence of oxygen, bacteria utilize various oxidized substances, e.g. nitrate, sulphate, metallic (hydr)oxides, humic substance, as terminal electron acceptors (TEAs) to fuel anaerobic XOCs degradation. Although there have been increasing anaerobic biodegradation studies focusing on species identification, degrading pathways, community dynamics, systematic reviews on the underlying mechanism of anaerobic contaminants removal from the perspective of electron flow are limited. In this review, we provide the insight on anaerobic biodegradation from electrons aspect - electron production, transport, and consumption. The mechanism of the coupling between TEAs reduction and pollutants degradation is deconstructed in the level of community, pure culture, and cellular biochemistry. Hereby, relevant strategies to promote anaerobic biodegradation are proposed for guiding to an efficient XOCs bioremediation.

Effective removal of antibiotic resistance genes and potential links with archaeal communities during vacuum-type composting and positive-pressure composting.

As a major reservoir of antibiotics, animal manure contributes a lot to the augmented environmental pressure of antibiotic resistance genes (ARGs). This might be the first study to explore the effects of different ventilation types on the control of ARGs and to identify the relationships between archaeal communities and ARGs during the composting of dairy manure. Several ARGs were quantified via Real-time qPCR and microbial communities including bacteria and archaea were analyzed by High-throughput sequencing during vacuum-type composting (VTC) and positive-pressure composting (PPC). The total detected ARGs and class I integrase gene (intI1) under VTC were significantly lower than that under PPC during each stage of the composting (p<0.001). The relative abundance of potential human pathogenic bacteria (HPB) which were identified based on sequencing information and correlation analysis decreased by 74.6% and 91.4% at the end of PPC and VTC, respectively. The composition of archaeal communities indicated that methane-producing archaea including Methanobrevibacter, Methanocorpusculum and Methanosphaera were dominant throughout the composting. Redundancy analysis suggested that Methanobrevibacter and Methanocorpusculum were positively correlated with all of the detected ARGs. Network analysis determined that the possible hosts of ARGs were different under VTC and PPC, and provided new sights about potential links between archaea and ARGs. Our results showed better performance of VTC in reducing ARGs and potential HPB and demonstrated that some archaea could also be influential hosts of ARGs, and caution the risks of archaea carrying ARGs.

Genetic diversity of diazotrophs and total bacteria in the phyllosphere of Pyrus serotina, Prunus armeniaca, Prunus avium, and Vitis vinifera.

The phyllosphere, which supports a large number of microorganisms, represents the interface between the aboveground parts of plants and air. In this study, four nifH clone libraries were constructed from the phyllosphere of Pyrus serotina (L), Vitis vinifera (P), Prunus armeniaca (X), and Prunus avium (Y). Clones related to Skermanella (L, 12.1%; X, 15.6%; Y, 62.5%; P 70.8%), Bradyrhizobium (X, 2.1%; P, 15.1%; L, 63.7%), Erwinia (X, 68.8%), Pseudomonas (L, 3.3%; P, 7.6%), and Chroococcidiopsis (P, 0.9%; L, 4.4%, X; 5.2%, Y; 19.6%) were present at high percentages, highlighting their critical role in contributing nitrogen to the phyllosphere ecosystem. The 16S rDNA sequence analysis suggested that phyllosphere-associated bacteria were affiliated with a wide range of taxa, encompassing members from Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Cyanobacteria, Tenericutes, and Deinococcus-Thermus. Additionally, the abundance of the nifH gene and 16S rDNA was assessed with quantitative PCR. The number of copies of nifH and 16S rDNA ranged from 1.14 × 103 to 1.49 × 104 and from 3.72 × 106 to 7.02 × 107 copies/g fresh leaf sample, respectively. In conclusion, our work sheds light on the microbial communities of the phyllosphere that are important for plant growth. Moreover, we observed a unique composition of nitrogen-fixing bacteria in each phyllosphere sample, suggesting the existence of specific interactions between these functional microorganism and plants, which may provide information or be a reference for the development of bacterial fertilizers.

Transcriptomic Sequencing and Co-Expression Network Analysis on Key Genes and Pathways Regulating Nitrogen Use Efficiency in Myriophyllum aquaticum.

Massively input and accumulated ammonium is one of the main causes of eutrophication in aquatic ecosystems, which severely deteriorates water quality. Previous studies showed that one of the commonly used macrophytes, Myriophyllum aquaticum, was capable of not only withstanding ammonium of high concentration, but also efficiently assimilating extracellular ammonium to constitutive amino acids and proteins. However, the genetic mechanism regulating such efficient nitrogen metabolism in M. aquaticum is still poorly understood. Therefore, RNA-based analysis was performed in this study to understand the ammonium regulatory mechanism in M. aquaticum in response to various concentrations of ammonium. A total of 7721 genes were differentially expressed, of which those related to nitrogen-transport, assimilation, and remobilization were highly-regulated in response to various concentrations of ammonium. We have also identified transcription factors and protein kinases that were rapidly induced in response to ammonium, which suggests their involvement in ammonium-mediated signalling. Meanwhile, secondary metabolism including phenolics and anthocyanins biosynthesis was also activated in response to various concentrations of ammonium, especially at high ammonium concentrations. These results proposed a complex physiological and genetic regulation network related to nitrogen, carbohydrate, transcription factors, and secondary metabolism for nitrogen use efficiency in M. aquaticum.

Enhancement of facultative anaerobic denitrifying communities by oxygen release from roots of the macrophyte in constructed wetlands.

The radial oxygen loss (ROL) of wetland plants is a crucial factor that can influence the efficiency required for nitrogen (N) removal and microbial activities responsible for N removal in constructed wetlands (CWs). However, the shift of microbial community in different niches in response to ROL has been rarely studied. This study aims to unravel the link between the ROL and microbial response in sediment, water and rhizoplane by a surface flow CW planted with Myriophyllum aquaticum for treating high-strength swine wastewater. Ti3+-citrate colorimetric method demonstrated that M. aquaticum was a wetland species with a ROL of 0.019 mg/h/plant. Using quantitative polymerase chain reactions (qPCR) and high-throughput sequencing of microbial 16S rRNA gene, we demonstrated that the abundance of facultative anaerobic denitrifiers in the rhizoplane was the most of the three niches, that in the water (5-10 cm) was the less and that in the sediment was the least. Acinetobacter was enriched and dominated amongst denitrifiers in the water. Denitrifiers in the rhizoplane were mainly dominated by enriched Pseudomonas, Aeromonas, and Acinetobacter. The theoretical calculation of oxygen sources and consumptions indicated that water reaeration should support the oxygen demands for nitrification in the aerobic layer (0-5 cm), and the ROL could stimulate the growth of facultative anaerobic denitrifiers in the rhizoplane and water (5-10 cm) to achieve denitrification within CW systems.

Long- and short-chain AHLs affect AOA and AOB microbial community composition and ammonia oxidation rate in activated sludge.

Quorum sensing (QS) regulation of the composition of ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) communities and functions in wastewater treatment was investigated. Specifically, we explored the role of N-acyl-l-homoserine lactones (AHLs) in microbial community dynamics in activated sludge. On average, the specific ammonia-oxidising-rate increased from 1.6 to 2.8 mg NH4+-N/g MLSS/hr after treatment with long-chain AHLs for 16 days, and the addition of AHLs to sludge resulted in an increased number of AOA/AOB amoA genes. Significant differences were observed in the AOA communities of control and AHL-treated cultures, but not the AOB community. Furthermore, the dominant functional AOA strains of the Crenarchaeota altered their ecological niche in response to AHL addition. These results provide evidence that AHLs play an important role in mediating AOA/AOB microbial community parameters and demonstrate the potential for application of QS to the regulation of nitrogen compound metabolism in wastewater treatment.

Electron acceptors for anaerobic oxidation of methane drive microbial community structure and diversity in mud volcanoes.

Mud volcanoes (MVs) emit globally significant quantities of methane into the atmosphere, however, methane cycling in such environments is not yet fully understood, as the roles of microbes and their associated biogeochemical processes have been largely overlooked. Here, we used data from high-throughput sequencing of microbial 16S rRNA gene amplicons from six MVs in the Junggar Basin in northwest China to quantify patterns of diversity and characterize the community structure of archaea and bacteria. We found anaerobic methanotrophs and diverse sulfate- and iron-reducing microbes in all of the samples, and the diversity of both archaeal and bacterial communities was strongly linked to the concentrations of sulfate, iron and nitrate, which could act as electron acceptors in anaerobic oxidation of methane (AOM). The impacts of sulfate/iron/nitrate on AOM in the MVs were verified by microcosm experiments. Further, two representative MVs were selected to explore the microbial interactions based on phylogenetic molecular ecological networks. The sites showed distinct network structures, key species and microbial interactions, with more complex and numerous linkages between methane-cycling microbes and their partners being observed in the iron/sulfate-rich MV. These findings suggest that electron acceptors are important factors driving the structure of microbial communities in these methane-rich environments.

The Occurrence of Putative Nitric Oxide Dismutase (Nod) in an Alpine Wetland with a New Dominant Subcluster and the Potential Ability for a Methane Sink.

Recently, a new oxygenic pathway has been proposed based on the disproportionation of NO with putative NO dismutase (Nod). In addition to a new process in nitrogen cycling, this process provides ecological advantages for the degradation of substrates in anaerobic conditions, which is of great significance for wastewater treatment. However, the Nod distribution in aquatic environments is rarely investigated. In this study, we obtained the nod genes with an abundance of 2.38 ± 0.96 × 105 copies per gram of dry soil from the Zoige wetland and aligned the molecular characteristics in the corresponding Nod sequences. These Nod sequences were not only found existing in NC10 bacteria, but were also found forming some other clusters with Nod sequences from a WWTP reactor or contaminated aquifers. Moreover, a new subcluster in the aquifer-similar cluster was even dominant in the Zoige wetland and was named the Z-aquifer subcluster. Additionally, soils from the Zoige wetland showed a high potential rate (10.97 ± 1.42 nmol of CO2 per gram of dry soil per day) for nitrite-dependent anaerobic methane oxidation (N-DAMO) with low abundance of NC10 bacteria, which may suggest a potential activity of Nod in other clusters when considering the dominance of the Z-aquifer subcluster Nod. In conclusion, we verified the occurrence of Nod in an alpine wetland for the first time and found a new subcluster to be dominant in the Zoige wetland. Moreover, this new subcluster of Nod may even be active in the N-DAMO process in this alpine wetland, which needs further study to confirm.