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.

Efficient elimination of antibiotic resistome in livestock manure by semi-permeable membrane covered hyperthermophilic composting.

Livestock manure is a hotspot for antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs), and an important contributor to antibiotic resistance in non-clinical settings. This study investigated the effectiveness and potential mechanisms of a novel composting technology, semi-permeable membrane covered hyperthermophilic composting (smHTC), in removal of ARGs and MGEs in chicken manure. Results showed that smHTC was more efficient in removal of ARGs and MGEs (92% and 93%) compared to conventional thermophilic composting (cTC) (76% and 92%). The efficient removal in smHTC is attributed to direct or indirect negative effects caused by the high temperature, including reducing the involvement of bio-available heavy metals (HMs) in co-selection processes of antibiotic resistance, decreasing the bacterial abundance and diversity, suppressing the horizontal gene transfer and killing potential ARGs hosts. Overall, smHTC can efficiently remove the resistome in livestock manure, reducing the risk to crops and humans from ARGs residues in compost products.

Season and side-chain length affect the occurrences and behaviors of phthalic acid esters in wastewater treatment plants.

Emerging pollutants (EPs) are prevalent in aquatic environments globally. Researchers strive to understand their occurrence and behavior prior to their release into the environment. In this study, we examined five wastewater treatment plants (WWTPs), collected 50 wastewater samples and 10 sludge samples. We explored the sources and destinations of phthalic acid esters (PAEs) within these WWTPs using mass balance equations. Wastewater treatment diminished the frequency and concentration of PAEs, and decreased the fraction of short-chain PAEs. We confirmed the increased concentration of PAEs post-primary treatment and modified the mass balance equation. Calculations suggest that weaker "the mix" in winter than in summer and stronger sedimentation in winter than in summer resulted in high efficiency of PAEs removal by winter wastewater treatment. The mass flux of biodegradation was influenced by the combination of biodegradation efficiency and the strength of the particular type of PAEs collected, with no seasonal differences. Mass fluxes for sludge sedimentation were mainly influenced by season and were higher in winter than in summer. This study enhances our understanding of emerging pollutants in manual treatment facilities and offers insights for optimizing wastewater treatment methods for water professionals.

Non-negligible roles of upstream rivers in determining the antibiotic resistance genes community in an interconnected river-lake system (Dongting lake, China).

Emergence and spread of antibiotic resistance genes (ARGs) in lakes have been considered as a global health threat. However, a thorough understanding of the distribution patterns and ecological processes that shape the ARGs profile in interconnected river-lake systems remains largely unexplored. In this study, we collected paired water and sediment samples from a typical interconnected river-lake system, Dongting Lake in China, during both wet and dry seasons. Using high-throughput quantitative PCR, we investigated the spatial and temporal distribution of ARGs and the factors that influence them. A total of 8 major antibiotic classes and 10 mobile genetic elements were detected across the Dongting Lake basin. The unique hydrological characteristics of this interconnected river-lake system result in a relatively stable abundance of ARGs across different seasons and interfaces. During the wet season, deterministic processes dominated the assembly of ARGs, allowing environmental factors, such as heavy metals, to serve as main driving forces of ARGs distribution. When the dry season arrived, variations in hydrological conditions and changes in ARGs sources caused stochastic processes to dominate the assembly of ARGs. Our findings provide valuable insights for understanding the ecological processes of ARGs in interconnected river-lake systems, emphasizing the necessity of upstream restoration and clarifying river-lake relationships to mitigate ARGs dissemination.

Electron Transfer in the Biogeochemical Sulfur Cycle.

Microorganisms are key players in the global biogeochemical sulfur cycle. Among them, some have garnered particular attention due to their electrical activity and ability to perform extracellular electron transfer. A growing body of research has highlighted their extensive phylogenetic and metabolic diversity, revealing their crucial roles in ecological processes. In this review, we delve into the electron transfer process between sulfate-reducing bacteria and anaerobic alkane-oxidizing archaea, which facilitates growth within syntrophic communities. Furthermore, we review the phenomenon of long-distance electron transfer and potential extracellular electron transfer in multicellular filamentous sulfur-oxidizing bacteria. These bacteria, with their vast application prospects and ecological significance, play a pivotal role in various ecological processes. Subsequently, we discuss the important role of the pili/cytochrome for electron transfer and presented cutting-edge approaches for exploring and studying electroactive microorganisms. This review provides a comprehensive overview of electroactive microorganisms participating in the biogeochemical sulfur cycle. By examining their electron transfer mechanisms, and the potential ecological and applied implications, we offer novel insights into microbial sulfur metabolism, thereby advancing applications in the development of sustainable bioelectronics materials and bioremediation technologies.

Understanding Salinity-Driven Modulation of Microbial Interactions: Rhizosphere versus Edaphic Microbiome Dynamics.

Soil salinization poses a global threat to terrestrial ecosystems. Soil microorganisms, crucial for maintaining ecosystem services, are sensitive to changes in soil structure and properties, particularly salinity. In this study, contrasting dynamics within the rhizosphere and bulk soil were focused on exploring the effects of heightened salinity on soil microbial communities, evaluating the influences shaping their composition in saline environments. This study observed a general decrease in bacterial alpha diversity with increasing salinity, along with shifts in community structure in terms of taxa relative abundance. The size and stability of bacterial co-occurrence networks declined under salt stress, indicating functional and resilience losses. An increased proportion of heterogeneous selection in bacterial community assembly suggested salinity's critical role in shaping bacterial communities. Stochasticity dominated fungal community assembly, suggesting their relatively lower sensitivity to soil salinity. However, bipartite network analysis revealed that fungi played a more significant role than bacteria in intensified microbial interactions in the rhizosphere under salinity stress compared to the bulk soil. Therefore, microbial cross-domain interactions might play a key role in bacterial resilience under salt stress in the rhizosphere.

Transfer and accumulation of antibiotic resistance genes and bacterial pathogens in the mice gut due to consumption of organic foods.

Over the last few decades, organic food demand has grown largely because of increasing personal health concerns. Organic farming introduces antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) into foods. However, potential effects of organic foods on the gut microbiome and ARGs have been overlooked. Using high-throughput quantitative PCR and 16S rRNA high-throughput sequencing technology, we examined 132 ARGs from major classes, eight transposase genes, universal class I integron-integrase gene (intI), clinical class I integron-integrase gene (cintI), and the bacterial community in mouse gut after 8 weeks with an either organic or inorganic lettuce and wheat diet. A total of 8 types of major ARGs and 10 mobile genetic elements (MGEs) were detected in mice gut, including tetracycline, multidrug, sulfonamide, aminoglycoside, beta-lactamase, chloramphenicol, MLSB and vancomycin resistance genes. We found that abundance and diversity of ARGs, mobile gene elements, and potential ARB in the gut increased with time after consumption of organic foods, whereas no significant changes were observed in inorganic treated groups. Moreover, MGEs, including IS613, Tp614 and tnpA_03 were found to play an important role in regulating ARG profiles in the gut microbiome following consumption of organic foods. Importantly, feeding organic food increased the relative abundance of the potentially antibiotic-resistant pathogens, Bacteroides and Streptococcus. Our results confirm that there is an increasing risk of ARGs and ARB in the gut microbiome, which highlights the importance of organic food industries taking into account the potential accumulation and transmission of ARGs as a risk factor.

Rapid start-up of an innovative pilot-scale staged PN/A continuous process for enhanced nitrogen removal from mature landfill leachate via robust NOB elimination and efficient biomass retention.

The start-up and stable operation of partial nitritation-anammox (PN/A) treatment of mature landfill leachate (MLL) still face challenges. This study developed an innovative staged pilot-scale PN/A system to enhance nitrogen removal from MLL. The staged process included a PN unit, an anammox upflow enhanced internal circulation biofilm (UEICB) reactor, and a post-biofilm unit. Rapid start-up of the continuous flow PN process (full-concentration MLL) was achieved within 35 days by controlling dissolved oxygen and leveraging free ammonia and free nitrous acid to selectively suppress nitrite-oxidizing bacteria (NOB). The UEICB was equipped with an annular flow agitator combined with the enhanced internal circulation device of the guide tube, which achieved an efficient enrichment of Candidatus Kuenenia in the biofilm (relative abundance of 33.4 %). The nitrogen removal alliance formed by the salt-tolerant anammox bacterium (Candidatus Kuenenia) and denitrifying bacteria (unclassified SBR1031 and Denitratisoma) achieved efficient nitrogen removal of UEICB (total nitrogen removal percentage: 90.8 %) and at the same time effective treatment of the refractory organic matter (ROM). The dual membrane process of UEICB fixed biofilm combined with post-biofilm is effective in sludge retention, and can stably control the effluent suspended solids (SS) at a level of less than 5 mg/L. The post-biofilm unit ensured that effluent total nitrogen (TN) remained below the 40 mg/L discharge standard (98.5 % removal efficiency). Compared with conventional nitrification-denitrification systems, the staged PN/A process substantially reduced oxygen consumption, sludge production, CO2 emissions and carbon consumption by 22.8 %, 67.1 %, 87.1 % and 87.1 %, respectively. The 195-day stable operation marks the effective implementation of the innovative pilot-scale PN/A process in treating actual MLL. This study provides insights into strategies for rapid start-up, robust NOB suppression, and anammox biomass retention to advance the application of PN/A in high-ammonia low-carbon wastewater.

The interaction of climate, plant, and soil factors drives putative soil fungal pathogen diversity and community structure in dry grasslands.

Soil pathogens play important roles in shaping soil microbial diversity and controlling ecosystem functions. Though climate and local environmental factors and their influences on fungal pathogen communities have been examined separately, few studies explore the relative contributions of these factors. This is particularly crucial in eco-fragile regions, which are more sensitive to environmental changes. Herein we investigated the diversity and community structure of putative soil fungal pathogens in cold and dry grasslands on the Tibetan Plateau, using high-throughput sequencing. The results showed that steppe soils had the highest diversity of all pathogens and plant pathogens; contrastingly, meadow soils had the highest animal pathogen diversity. Structural equation modelling revealed that climate, plant, and soil had similar levels of influence on putative soil fungal pathogen diversity, with total effects ranging from 52% to 59% (all p < 0.001), with precipitation exhibiting a stronger direct effect than plant and soil factors. Putative soil fungal pathogen community structure gradually changed with desert, steppe, and meadow, and was primarily controlled by the interactions of climate, plant, and soil factors rather than by distinct factors individually. This finding contrasts with most studies of soil bacterial and fungal community structure, which generally report dominant roles of individual environmental factors.

N2O emission reduction in the biological nitrogen removal process for wastewater with low C/N ratios: mechanisms and strategies.

Urban wastewater, as the main influent type of Waste Water Treatment Plants (WWTPs), has the characteristic of low carbon to nitrogen ratio (C/N). In the biological nitrogen removal (BNR) process, insufficient carbon source often affects the nitrogen removal efficiency and leads to more N2O emissions. We review recent researches on N2O emissions in the BNR process of wastewater with low C/N. The availability of carbon sources affects heterotrophic denitrification (HD) and autotrophic nitrification/denitrification processes, which are the main reasons for N2O emissions in BNR. For the sustainable development of BNR in WWTPs, we introduce strategies suitable for reducing N2O emissions in the BNR process of low C/N wastewater from two aspects: traditional process innovation and new process development. These strategies mainly include carbon source addition, adjustment of aeration strategy, optimization of oxidation ditch and biofilm facilities, and application of Anammox related processes. In the future, it is still necessary to further deepen this research direction through the normalization of N2O emission quantification standards, exploration of N2O metabolism mechanisms, assessment of environmental effects of emission reduction strategies, and practical application of new processes.

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.

Temporal and spatial changes of water quality in intensively developed urban rivers and water environment improvement: a case study of the Longgang River in Shenzhen, China.

The water quality status, spatial and temporal change processes, and water environment improvement process of urban rivers are valuable lessons to be learned under the sustainable development strategy. This study aims to reveal the water environment improvement process of intensively developed urban rivers, elucidate the spatial and temporal distribution characteristics of major pollutants, and provide recommendations for their water environment management. Water quality data from eight monitoring sites (2007-2020) in the Longgang River basin in Shenzhen, China, and comprehensive pollution index method (CPI), modified comprehensive pollution index method (M-CPI), and Pearson correlation analysis method were used for comprehensive analysis. The study shows that TN, TP, NH3-N, and COD have the greatest influence on the water quality of Longgang River, with the average pollution contribution of 53.39%, 14.49%, 11.66%, and 4.92%, in order. In 2015-2020, the water quality of the main stream of the Longgang River in the wet season was worse than that in the dry season, while the water quality of the tributaries Dingshan River and the Huangsha River in the dry season was worse than the wet season. The spatial distribution characteristics based on M-CPI indicate that the water quality of the lower reaches of Longgang River, the tributaries Dingshan River and Huangsha River, is relatively poor. In addition, the water environment improvement process of Longgang River can be divided into 3 stages: engineering stage (2007-2013, rating changed from heavily polluted to basically qualified), bottleneck stage (2013-2017, rating fluctuated slightly above and below basically qualified), and ecological restoration stage (2017-2020, rating reached qualified in 2019).

The effects, mechanisms, and applications of sulfide as both an inhibitor and electron donor in novel biological nitrogen removal process.

The impacts of sulfide on biological nitrogen removal have been investigated repeatedly, but the impacts in terms of the removal technologies have not been systematically organized and discussed yet. This review recapped the dual character of sulfide in novel biological nitrogen removal and proposed the coupling mechanisms of nitrogen removal and sulfide interaction. The dual character of sulfide was basically divided into the advantage as electron donors and the disadvantage as cytotoxic agent to wide bacteria. The positive character of sulfide has been utilized for improving the performances of denitrification and anaerobic ammonium oxidation in laboratory and polit scales. Intriguingly, the sulfide cytotoxicity was feasibly turned into the profit which was to trigger the partial nitrification by selectively inhibiting ammonia oxidizing bacteria and nitrite oxidizing bacteria. Thus, this productive conversion dramatically promoted the importance of sulfide in sewage treatment. In order to maximize the favorable aspect of sulfide utilization, it was crucial to manage the sulfide concentration for fear of the side reactions with untargeted substances. Furthermore, S/N ratio in sewage may be the keystone that decides if sulfide benefits biological nitrogen removal. In sum, our work can facilitate the dialectical development of effective strategies for sulfide utilization in biological nitrogen removal.

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.

In Situ Enhanced Yields of Microbial Nanowires: The Key Role of Environmental Stress.

The conductive microbial nanowires of Geobacter sulfurreducens serve as a model for long-range extracellular electron transfer (EET), which is considered a revolutionary "green" nanomaterial in the fields of bioelectronics, renewable energy, and bioremediation. However, there is no efficient pathway to induce microorganisms to express a large amount of microbial nanowires. Here, several strategies have been used to successfully induce the expression of microbial nanowires. Microbial nanowire expression was closely related to the concentration of electron acceptors. The microbial nanowire was around 17.02 µm in length, more than 3 times compared to its own length. The graphite electrode was used as an alternative electron acceptor by G. sulfurreducens, which obtained a fast start-up time of 44 h in microbial fuel cells (MFCs). Meanwhile, Fe(III) citrate-coated sugarcane carbon and biochar were prepared to test the applicability of these strategies in the actual microbial community. The unsatisfied EET efficiency between c-type cytochrome and extracellular insoluble electron receptors promoted the expression of microbial nanowires. Hence, microbial nanowires were proposed to be an effective survival strategy for G. sulfurreducens to cope with various environmental stresses. Based on this top-down strategy of artificially constructed microbial environmental stress, this study is of great significance for exploring more efficient methods to induce microbial nanowires expression.

Quorum sensing responses of activated sludge to free nitrous acid: Zoogloea deformation, AHL redistribution, and microbiota acclimatization.

Free nitrous acid (FNA) has been widely employed for improvement of wastewater management by altering sludge characteristic and function based on its polymer lysing and biocidal capacity. Sludge characteristic and function are commonly considered as the joint consequence of microbial individual behaviors and quorum sensing (QS) involved collective behaviours, but the role of the latter in FNA treatment was still as-yet-unidentified and addressed in this research. The results of sludge morphology and component characterized FNA-induced zoogloea deformation, including inner cell exposure, half of extracellular polymeric substances (EPS) reduction and adsorption site depletion. During zoogloea deformation, four acyl-homoserine lactones (AHLs), including C4-HSL, C8-HSL, C10-HSL and C12-HSL, transferred inward of microbiota, and their total contents reduced by 66% because of depressed signal production, augmented decomposer and recognition. Transcriptome analysis revealed that differentially expressed QS driven by AHL redistribution facilitated microbiota acclimatization including cellular motility and hydrolase synthesis for EPS consumption. Boosted motility may favor escaping from stress spot and moderating intercellular acidity based on cell motility test. Feasible EPS consumption provided nutrition for heterotrophic metabolisms testified by pure culture with EPS as sole nutrition. Our work thus comprehensively revealed QS behaviours responding to FNA and deepened the understanding to FNA treatment performance in wastewater management.

Phylogenetic distance affects the artificial microbial consortia's effectiveness and colonization during the bioremediation of polluted soil with Cr(VI) and atrazine.

Soils co-contaminated with heavy metals and organic pollutants are common and threaten the natural environment and human health. Although artificial microbial consortia have advantages over single strains, the mechanism affecting their effectiveness and colonization in polluted soils still requires determination. Here, we constructed two kinds of artificial microbial consortia from the same or different phylogenetic groups and inoculated them into soil co-contaminated with Cr(VI) and atrazine to study the effects of phylogenetic distance on consortia effectiveness and colonization. The residual concentrations of pollutants demonstrated that the artificial microbial consortium from different phylogenetic groups achieved the highest removal rates of Cr(VI) and atrazine. The removal rate of 400 mg/kg atrazine was 100%, while that of 40 mg/kg Cr(VI) was 57.7%. High-throughput sequence analysis showed that the soil bacterial negative correlations, core genera, and potential metabolic interactions differed among treatments. Furthermore, artificial microbial consortia from different phylogenetic groups had better colonization and a more significant effect on the abundance of native core bacteria than consortia from the same phylogenetic group. Our study highlights the importance of phylogenetic distance on consortium effectiveness and colonization and offers insight into the bioremediation of combined pollutants.

Quorum sensing improves the plant growth-promoting ability of Stenotrophomonas rhizophila under saline-alkaline stress by enhancing its environmental adaptability.

Quorum sensing (QS) system has an essential function in plant growth-promoting rhizobacteria (PGPR) response to environmental stress and PGPR induction of plant tolerance to saline-alkaline stress. Nevertheless, there is a lack of understanding about how QS influences the growth-promoting effects of PGPR on plants. Stenotrophomonas rhizophila DSM14405T is a PGPR with a QS system, which can secrete diffusible signal factor (DSF), one of the QS signal molecules. In this study, we used the S. rhizophila wild type (WT) and an incompetent DSF production rpfF-knockout mutant strain to explore whether DSF-QS could affect the growth-promoting ability of PGPR in Brassica napus L. By measuring the seed germination rate, plant fresh weight, biomass, the total antioxidant capacity (T-AOC) level, and the content of chlorophyll in leaves, we found that DSF was unable to enhance the growth-promoting capacity of ΔrpfF and did not directly assist the plants in tolerating saline-alkaline stress. However, DSF aided S. rhizophila ΔrpfF in resisting stress during its effective period, and QS represents a continuous and precise regulatory mechanism. Altogether, our results show that DSF is helpful to improve the environmental adaptability and survival rate of S. rhizophila, thus indirectly improving the germination rate of seeds and helping plants grow under saline-alkaline stress. In this study, the mechanism of QS enhancing the environmental adaptability of PGPR was studied, which provided a theoretical basis for improving the application of PGPR to help plants better cope with saline-alkaline stress.

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.

Carbon footprint analysis and comprehensive evaluation of municipal wastewater treatment plants under different typical upgrading and reconstruction modes.

The issue of greenhouse gas (GHG) emissions resulting from the upgrading and reconstruction of municipal wastewater treatment plants (MWWTPs) along with improved water quality is receiving attention and research. There is an urgent need to explore the impact of upgrading and reconstruction on carbon footprint (CF) in order to address concerns that the upgrading and reconstruction will increase GHG emissions while improving water quality. Here we accounted for the CF of five MWWTPs in Zhejiang Province, China, before and after three different upgrading and reconstruction models - "Improving quality and efficiency" ("Mode I"), "Upgrading and renovation" ("Mode U") and "Improving quality and efficiency plus Upgrading and renovation" ("Mode I plus U"). The upgrading and reconstruction was found to not necessarily result in more GHG emissions. In contrast, the "Mode I" had a more significant advantage in terms of CF reduction (1.82-12.6 % reduction in CF). Overall, the ratio of indirect emissions to direct emissions (indirect emissions/direct emissions) and the amount of GHG emitted per unit of pollutant removed (CFCOD、CFTN、CFTP) decreased, while both the carbon and energy neutral rates increased significantly (up to 33.29 % and 79.36 % respectively) after all three upgrading and reconstruction modes. In addition, the wastewater treatment efficiency and capacity are the main factors that affect the level of carbon emission. The results of this study can provide a calculation model that can be used for other similar MWWTPs during the upgrading and reconstruction processes. More importantly, it can provide a new research perspective as well as valuable information to revisit the impact of upgrading and reconstruction in MWWTPs on GHG emissions.