Confined water-encapsulated activated carbon for capturing short-chain perfluoroalkyl and polyfluoroalkyl substances from drinking water.

The global ecological crisis of perfluoroalkyl and polyfluoroalkyl substances (PFASs) in drinking water has gradually shifted from long-chain to short-chain PFASs; however, the widespread established PFAS adsorption technology cannot cope with the impact of such hydrophilic pollutants given the inherent defects of solid-liquid mass transfer. Herein, we describe a reagent-free and low-cost strategy to reduce the energy state of short-chain PFASs in hydrophobic nanopores by employing an in situ constructed confined water structure in activated carbon (AC). Through direct (driving force) and indirect (assisted slip) effects, the confined water introduced a dual-drive mode in the confined water-encapsulated activated carbon (CW-AC) and completely eliminated the mass transfer barrier (3.27 to 5.66 kcal/mol), which caused the CW-AC to exhibit the highest adsorption capacity for various short-chain PFASs (C-F number: 3-6) among parent AC and other adsorbents reported. Meanwhile, benefiting from the chain length- and functional group-dependent confined water-binding pattern, the affinity of the CW-AC surpassed the traditional hydrophobicity dominance and shifted toward hydrophilic short-chain PFASs that easily escaped treatment. Importantly, the ability of CW-AC functionality to directly transfer to existing adsorption devices was verified, which could treat 21,000 bed volumes of environment-related high-load (~350 ng/L short-chain PFAS each) real drinking water to below the World Health Organization's standard. Overall, our results provide a green and cost-effective in situ upgrade scheme for existing adsorption devices to address the short-chain PFAS crisis.

N-Decorated Main-Group MgAl2O4 Spinel: Unlocking Exceptional Oxygen Reduction Activity for Zn-Air Batteries.

The development of economical and efficient oxygen reduction reaction (ORR) catalysts is crucial to accelerate the widespread application rhythm of aqueous rechargeable zinc-air batteries (ZABs). Here, a strategy is reported that the modification of the binding energy for reaction intermediates by the axial N-group converts the inactive spinel MgAl2O4 into the active motif of MgAl2O4-N. It is found that the introduction of N species can effectively optimize the electronic configuration of MgAl2O4, thereby significantly reducing the adsorption strength of *OH and boosting the reaction process. This main-group MgAl2O4-N catalyst exhibits a high ORR activity in a broad pH range from acidic and alkaline environments. The aqueous ZABs assembled with MgAl2O4-N shows a peak power density of 158.5 mW cm-2, the long-term cyclability over 2000 h and the high stability in the temperature range from -10 to 50 °C, outperforming the commercial Pt/C in terms of activity and stability. This work not only serves as a significant candidate for the robust ORR electrocatalysts of aqueous ZABs, but also paves a new route for the effective reutilization of waste Mg alloys.

2-Hydroxy-1,4-Naphthoquinone: A Promising Redox Mediator for Minimizing Dissolved Organic Nitrogen and Eutrophication Effects of Wastewater Effluent.

Researchers and engineers are committed to finding effective approaches to reduce dissolved organic nitrogen (DON) to meet more stringent effluent total nitrogen limits and minimize effluent eutrophication potential. Here, we provided a promising approach by adding specific doses of 2-hydroxy-1,4-naphthoquinone (HNQ) to postdenitrification bioreactors. This approach of adding a small dosage of 0.03-0.1 mM HNQ effectively reduced the concentrations of DON in the effluent (ANOVA, p < 0.05) by up to 63% reduction of effluent DON with a dosing of 0.1 mM HNQ when compared to the control bioreactors. Notably, an algal bioassay indicated that DON played a dominant role in stimulating phytoplankton growth, thus effluent eutrophication potential in bioreactors using 0.1 mM HNQ dramatically decreased compared to that in control bioreactors. The microbe-DON correlation analysis showed that HNQ dosing modified the microbial community composition to both weaken the production and promote the uptake of labile DON, thus minimizing the effluent DON concentration. The toxic assessment demonstrated the ecological safety of the effluent from the bioreactors using the strategy of HNQ addition. Overall, HNQ is a promising redox mediator to reduce the effluent DON concentration with the purpose of meeting low effluent total nitrogen levels and remarkably minimizing effluent eutrophication effects.

Carbon Source in Tertiary Denitrification Regulates Dissolved Organic Nitrogen in Wastewater Effluent.

With global eutrophication and increasingly stringent nitrogen discharge restrictions, dissolved organic nitrogen (DON) holds considerable potential to upgrade advanced wastewater denitrification because of its large contribution to low-nitrogen effluents and stronger stimulation effect for algae. Here, we show that DON from the postdenitrification systems dominates effluent eutrophication potential under different carbon sources. Methanol resulted in significantly lower DON concentrations (0.84 ± 0.03 mg/L) compared with the total nitrogen removal-preferred acetate (1.11 ± 0.02 mg/L) (p < 0.05, ANOVA). With our well-developed mathematical model (R2 = 0.867-0.958), produced DON instead of shared (persist in both influent and effluent) and/or removed DON was identified as the key component for effluent DON variation (Pearson r = 0.992, p < 0.01). The partial least-squares path modeling analysis showed that it is the microbial community (r = 0.947, p < 0.01) rather than the predicted metabolic functions (r = 0.040, p > 0.1) that affected produced DON. Carbon sources rebuild the microorganism-DON interaction by affecting the structure of microbial communities with different abilities to generate and recapture produced DON to finally regulate effluent DON. This study revalues the importance of carbon source selection and overturns the current rationality of pursuing only the total nitrogen removal efficiency by emphasizing DON.

Microbial Transformation of Dissolved Organic Sulfur during the Oxic Process in 47 Full-Scale Municipal Wastewater Treatment Plants.

Dissolved organic sulfur (DOS) is a significant part of effluent organic matter of wastewater treatment plants (WWTPs) and poses a potential ecological risk for receiving waters. However, the oxic process is a critical unit of biological wastewater treatment for microorganisms performing organic matter removal, wherein DOS transformation and its mechanism are poorly understood. This study investigated the transformation of DOS during the oxic process in 47 full-scale municipal WWTPs across China from molecular and microbial aspects. Surprisingly, evident differences in DOS variations (ΔDOS) separated sampled WWTPs into two groups: 28 WWTPs with decreased DOS concentrations in effluents (ΔDOS < 0) and 19 WWTPs with increased DOS (ΔDOS > 0). These two groups also presented differences in DOS molecular characteristics: higher nitrogen/carbon (N/C) ratios (0.030) and more peptide-like DOS (8.2%) occurred in WWTPs with ΔDOS > 0, implying that peptide-like DOS generated from microbes contributed to increased DOS in effluents. Specific microbe-DOS correlations (Spearman correlation, p < 0.05) indicated that increased effluent DOS might be explained by peptide-like DOS preferentially being produced during copiotrophic bacterial growth and accumulating due to less active cofactor metabolisms. Considering the potential environmental issues accompanying DOS discharge from WWTPs with ΔDOS > 0, our study highlights the importance of focusing on the transformation and control of DOS in the oxic process.

Systemic LPS-induced microglial activation results in increased GABAergic tone: A mechanism of protection against neuroinflammation in the medial prefrontal cortex in mice.

Neuroinflammation with excess microglial activation and synaptic dysfunction are early symptoms of most neurological diseases. However, how microglia-associated neuroinflammation regulates synaptic activity remains obscure. We report here that acute neuroinflammation induced by intraperitoneal injection of lipopolysaccharide (LPS) results in cell-type-specific increases in inhibitory postsynaptic currents in the glutamatergic, but not the GABAergic, neurons of medial prefrontal cortex (mPFC), coinciding with excessive microglial activation. LPS causes upregulation in levels of GABAAR subunits, glutamine synthetase and vesicular GABA transporter, and downregulation in brain-derived neurotrophic factor (BDNF) and its receptor, pTrkB. Blockage of microglial activation by minocycline ameliorates LPS-induced abnormal expression of GABA signaling-related proteins and activity of synaptic and network. Moreover, minocycline prevents the mice from LPS-induced aberrant behavior, such as a reduction in total distance and time spent in the centre in the open field test; decreases in entries into the open arm of elevated-plus maze and in consumption of sucrose; increased immobility in the tail suspension test. Furthermore, upregulation of GABA signaling by tiagabine also prevents LPS-induced microglial activation and aberrant behavior. This study illustrates a mode of bidirectional constitutive signaling between the neural and immune compartments of the brain, and suggests that the mPFC is an important area for brain-immune system communication. Moreover, the present study highlights GABAergic signaling as a key therapeutic target for mitigating neuroinflammation-induced abnormal synaptic activity in the mPFC, together with the associated behavioral abnormalities.

Advances in Single-Cell Toxicogenomics in Environmental Toxicology.

The toxicity evaluation system of environmental pollutants has undergone numerous changes due to the application of new technologies. Single-cell toxicogenomics is rapidly changing our view on environmental toxicology by increasing the resolution of our analysis to the level of a single cell. Applications of this technology in environmental toxicology have begun to emerge and are rapidly expanding the portfolio of existing technologies and applications. Here, we first summarized different methods involved in single-cell isolation and amplification in single-cell sequencing process, compared the advantages and disadvantages of different methods, and analyzed their development trends. Then, we reviewed the main advances of single-cell toxicogenomics in environmental toxicology, emphatically analyzed the application prospects of this technology in identifying the target cells of pollutants in early embryos, clarifying the heterogeneous response of cell subtypes to pollutants, and finding pathogenic bacteria in unknown microbes, and highlighted the unique characteristics of this approach with high resolution, high throughput, and high specificity by examples. We also offered a prediction of the further application of this technology and the revolution it brings in environmental toxicology. Overall, these advances will provide practical solutions for controlling or mitigating exogenous toxicological effects that threaten human and ecosystem health, contribute to improving our understanding of the physiological processes affected by pollutants, and lead to the emergence of new methods of pollution control.

Novel insight into dissolved organic nitrogen (DON) transformation along wastewater treatment processes with special emphasis on endogenous-source DON.

Knowledge of endogenous-source dissolved organic nitrogen (esDON) produced in wastewater treatment processes is critical for evaluating its potential impacts on receiving waters because esDON is a recognized concern, as it causes eutrophication. However, differentiating esDON from influent residual DON in real wastewater is always a challenge. Here, we deciphered esDON information in DON transformation processes along a full-scale wastewater treatment train by combining multiple chemometric tools with ion-mobility separation quadrupole time-of-flight mass spectrometry (IMS-QTOF MS) analyses. In total, DON became more refractory and compact with shorter carbon chains and fewer nitrogen atoms, and esDON composed a nonnegligible fraction that dominated DON transformation and characteristics. New esDON produced in treatment processes constituted a crucial part (>35.5%) of wastewater DON, and its contributions to wastewater DON are augmented along the train. Evidence of molecular conformations further confirmed dominant roles of esDON in DON characteristics. Moreover, esDON participated in 46.7% of core biochemical reaction networks, explaining the importance of esDON in DON transformation. Our study offers a tool to gain esDON characteristics and transformation mechanisms, and highlights the importance to control esDON for alleviating adverse influences from DON in receiving waters.

Acute neuroinflammation increases excitability of prefrontal parvalbumin interneurons and their functional recruitment during novel object recognition.

There is an emerging body of literature suggesting that unlike the chronic neuroinflammatory response, acute neuroinflammation is self-regulated and is beneficial for central nervous system homeostasis and cognitive integrity. However, the neurophysiological alterations upon acute neuroinflammation and their implications on cognitive function remain poorly understood. In the present study, we reliably established a mouse model of acute and self-limiting neuroinflammation by administering a single intraperitoneal injection of low-dose lipopolysaccharide, which induced reversible sickness behavior and increased pro-inflammatory cytokine expression in the medial prefrontal cortex (mPFC). During acute neuroinflammation, fast-spiking parvalbumin-expressing interneurons (PV interneurons) in the mPFC exhibited a hyperexcitable phenotype exemplified by increased input resistance, decreased rheobase current, and a higher frequency of action potentials. Furthermore, PV interneurons in the prelimbic subregion of the mPFC were excessively recruited into circuits supporting novel object recognition memory, which remained intact after acute neuroinflammation. Together, our findings suggest that alterations in PV neuronal excitability resulting from acute neuroinflammation may mediate neuronal recruitment and confer a beneficial outcome on functional integrity of NOR circuit in the mPFC.

A New Method for Analyzing Aero-Optical Effects with Transient Simulation.

Aero-optical effects reduce the accuracy of optical sensors on high-speed aircraft. Current research usually focuses on light refraction caused by large-scale density structures in turbulence. A method for analyzing photon energy scattering caused by micro-scale structures is proposed in this paper, which can explain the macro image distortion caused by moving molecules in inhomogeneous airflow. Quantitative analysis of the propagation equation indicates that micro-scale structures may contribute more to the wavefront distortion than the widely considered large-scale structures. To analyze the micro mechanism of aero-optical effects, a transient simulator is designed based on the scaling model of transient distorted wavefronts and the artificial vortex structure. The simulation results demonstrate that correct aero-optical phenomena can be obtained from the micro mechanism of photon energy scattering. Examples of using the transient simulator to optimize the parameters of the star sensor on a hypersonic vehicle are provided. The proposed analysis method for micro-scale structures provides a new idea for studying the aero-optical effects.

Effect of Solids Retention Time on Effluent Dissolved Organic Nitrogen in the Activated Sludge Process: Studies on Bioavailability, Fluorescent Components, and Molecular Characteristics.

Wastewater-derived dissolved organic nitrogen (DON) should be minimized by municipal wastewater treatment plants (MWWTPs) to reduce its potential impact on receiving waters. Solids retention time (SRT) is a key control parameter for the activated sludge (AS) process; however, knowledge of its impact on effluent DON is limited. This study investigated the effect of SRT on the bioavailability, fluorescent components, and molecular characteristics of effluent DON in the AS process. Four lab-scale AS reactors were operated in parallel at different SRTs (5, 13, 26, and 40 days) for treatment of primary treated wastewater collected from an MWWTP. Results showed the positive effect of prolonged SRT on DON removal. AS reactors during longer SRTs, however, cannot sequester the bioavailable DON (ABDON) and occasionally contribute to greater amounts of ABDON in the effluents. Consequently, effluent DON bioavailability increased with SRT ( R2 = 0.619, p < 0.05, ANOVA). Analysis of effluent DON fluorescent components and molecular characteristics indicated that the high effluent DON bioavailability observed at long SRTs is contributed by the production of microbially derived nitrogenous organics. The results presented herein indicate that operating an AS process with a longer SRT cannot control the DON forms that readily stimulate algal growth.

Removal Characteristics of Dissolved Organic Nitrogen and Its Bioavailable Portion in a Postdenitrifying Biofilter: Effect of the C/N Ratio.

Addition of external carbon sources to postdenitrification biofilters (DNFs) is frequently used in municipal wastewater treatment plants to enhance dissolved inorganic nitrogen removal. However, little is known about its influence on the removal of dissolved organic nitrogen (DON). This study investigated the effect of the carbon-to-nitrogen (C/N) ratio (3, 4, 5, and 6) on the removal characteristics of DON and bioavailable DON (ABDON) in the pilot-scale DNFs treating real secondary effluent. Results showed that DNFs effluent DON accounted for 31.2-39.8% of the effluent total nitrogen. The maximum effluent DON and ABDON concentrations both occurred in DNF operated at a C/N ratio of 3. There was no significant difference in effluent DON concentrations in DNFs at C/N ratios of 4, 5, and 6; however, effluent ABDON and DON bioavailability significantly decreased with C/N ratios (p < 0.05, t-test). According to the chemical composition analysis, effluent DON at high C/N ratios tends to contain less % molecular weight < 1 kDa nitrogenous organic compounds and proteins/amino sugars-like nitrogenous organic formulas, which is likely responsible for its low bioavailability. Overall, this study indicates the benefit of a high C/N ratio during the DNF process in terms of controlling the DON forms that readily stimulate algal growth.

Biochar carrier application for nitrogen removal of domestic WWTPs in winter: challenges and opportunities.

Biofilm processes have a better nitrogen removal ability than traditional activated sludge at low temperatures (< 15 °C). Many biofilm processes, as well as integrated biofilm and activated sludge processes, are potential and realizable nitrogen removal upgrading methods for domestic wastewater treatment plants (WWTPs). Therefore, biofilm packing material is attractive for domestic WWTP upgrading and reconstruction in winter. For a half decade, researchers have successfully applied activated carbon to biochar as biofilm carrier in the wastewater treatment field. Biochar, as a biostable soil amendment with pores and crevices on its surface, has been applied in the soil-plant system, which promoted the adsorption of NH4+ and NO3-, decreased N2O emission, transcriptional level of narG, nxrA, and nirS, and changed the microbial community composition for better nitrogen removal. However, in the field of wastewater treatment, the study of biochar-packed process is merely in the laboratory stage of simulated wastewater, which deserves further research in the future. In this mini review, we will discuss the performances of different processes at low temperatures, the related mechanism of the biochar-packed process for nitrogen removal, and other potential applications of biochar carriers.

Intermittent polarization: A promising strategy for microbial electricity driven reduction of DOM toxicity in actual industrial wastewater.

Dissolved organic matter (DOM) in actual industrial wastewater comprises various compounds that trigger toxicity in aquatic organisms; thus, advanced treatment for reducing DOM toxicity is urgently needed to ensure safe effluent discharge. Herein, we successfully reduced the toxicity of DOM in actual industrial wastewater without external chemical addition by applying intermittent polarization to electrochemical bioreactors. The bioreactor operated under intermittent polarization effectively reduced the toxicity of DOM by 76.7 %, resulting in the toxicity of effluent DOM (determined by malformation rate of zebrafish larvae) reaching less than 3.5 %. Notably, DOM compounds with high double-bond equivalence (DBE ≥ 8) were identified as the key components responsible for the toxicity of DOM through ultrahigh-resolution mass spectrometry analysis. Insight into microbe-DOM interactions revealed that intermittent polarization promoted the microbial consumption of high-DBE components of DOM by both affecting microbial composition (ß = -0.5421, p < 0.01) and function (ß = -0.4831, p < 0.01), thus regulating effluent DOM toxicity. The study findings demonstrate that intermittent polarization is a promising strategy for microbial electricity-driven reduction of DOM toxicity in actual industrial wastewater to meet the increasing safety requirements of receiving waters.

Dissolved organic nitrogen depresses the expected outcome of wastewater treatment upgrading on effluent eutrophication potential mitigation: Molecular mechanistic insight.

Continuously tightening total nitrogen (TN) discharge standards in wastewater treatment plants is a common practice worldwide to mitigate eutrophication. However, given the different bioavailability of effluent dissolved organic nitrogen (DON) and inorganic nitrogen, a great inefficiency of the TN-targeted upgrading might be hidden because of the poor understanding of its impact on effluent eutrophication potential mitigation. Here we show that the tightening TN discharge standards could only considerably promote inorganic nitrogen removal, however, DON concentrations remained constant across different effluent TN levels (p > 0.05, Kruskal-Wallis test). Surprisingly, restricting TN in turn increases the reactivity of DON molecules owing to the accumulation of produced DON by acting on the key biotic and abiotic transformation reactions. The difficulty of removing DON and the increased DON reactivity during wastewater treatment upgrading contribute to the practical elimination effect of effluent eutrophication potential exhibiting lower than expected. This work challenges the rationality of the prevailing pursuit for extreme-low TN discharge, calling for shifting the focus of wastewater treatment upgrading towards the more fundamental eutrophication-targeted perspective.

Long-term impact of nano zero-valent iron on methanogenic activity, microbial community structure, and transcription activity in anaerobic wastewater treatment system.

Nano zero-valent iron (NZVI) is commonly used in industrial wastewater treatment. However, its long-term impact mechanisms of metabolization in anaerobic systems are not well understood. This study investigated the effects of long-term and continuous addition of NZVI on methanogenic activity, microbial community, and transcription activity. The results demonstrated that low levels of NZVI (1000 mg/L) induced inhibition of methanogenesis after 80 days, while high levels of NZVI (5000 mg/L) immediately led to a sharp decrease of cumulative methane production and chemical oxygen demand removal, which arrived at a steady state (14.4 % of control and 17 %) after 30 days. NZVI adversely affected cell viability, adenosine triphosphate production, and fatty acid evolution of cell membranes played a crucial role in resisting chronic NZVI toxicity. Moreover, high NZVI levels hindered the transcription of key enzymes CoM and mcrA, while low NZVI levels maintained its high CoM and mcrA activity, but down-regulated the transcription of cdh and hdr. Besides, amino-utilizing bacteria was reduced under the high NZVI concentration, while low NZVI changed dominant genus with potential protein hydrolysis function from Candidatus Cloacamonas to Sedimentibacter. These results provide a guideline for proper NZVI utilization in wastewater treatment.

Physical exercise mediates cortical synaptic protein lactylation to improve stress resilience.

Lactate is a critical metabolite during the body's adaption to exercise training, which effectively relieves anxiety-like disorders. The biological mechanism of lactate in the exercise-mediated anxiolytic effect has, however, not been comprehensively investigated. Here, we report that exercise-induced lactate markedly potentiates the lactylation of multiple synaptic proteins, among which synaptosome-associated protein 91 (SNAP91) is the critical molecule for synaptic functions. Both anatomical evidence and in vivo recording data showed that the lactylation of SNAP91 confers resilience against chronic restraint stress (CRS) via potentiating synaptic structural formation and neuronal activity in the medial prefrontal cortex (mPFC). More interestingly, exercise-potentiated lactylation of SNAP91 is necessary for the prevention of anxiety-like behaviors in CRS mice. These results collectively suggest a previously unrecognized non-histone lactylation in the brain for modulating mental functions and provide evidence for the brain's metabolic adaption during exercise paradigms.

Priority control sequence of 34 typical pollutants in effluents of Chinese wastewater treatment plants.

The identification of the priority control sequence of pollutants in effluents of wastewater treatment plants (WWTPs) has important implications for the management of water quality. This study chose 34 typical pollutants based on their representativeness and detection rates in municipal wastewater. The occurrence frequency and concentration of these pollutants in 168 Chinese WWTP effluents were measured at the national level. The data on in vitro toxicity (67 assays) and in vivo toxicity (216 species) for target pollutants were obtained from the public toxicity database and our experimental data. An environmental health prioritization index (EHPi) method was proposed to integrate the occurrence frequency, concentration, removal rate, and in vitro and in vivo toxicity to determine the priority control sequence of target pollutants. Ethynyl estradiol, 17ß-estradiol, estrone, diclofenac, and atrazine were the top 5 pollutants identified by the EHPi score. Several pollutants with high EHPi scores showed spatial differences. Besides the EHPi method which was from the single pollutant perspective, the combined toxicity of pollutants (300 pairs of binary combinations) was also measured based on in vitro toxicity assays to evaluate the key pollutants from the pollutant-pollutant interacting perspective. The pollutants (such as ofloxacin and acetaminophen) that could have significant synergetic effects with many other pollutants are worthy of prior attention. This study shed new light on the identification of the priority control sequence of pollutants in WWTP effluents. The results provide meaningful data for the effective management and control of wastewater water quality.

Identification and characterization of diverse isomers of per- and polyfluoroalkyl substances in Chinese municipal wastewater.

Linear and branched isomers of per- and polyfluoroalkyl substances (PFASs) are simultaneously present in the environment. However, isomer profiles of PFASs in municipal wastewater treatment plants (WWTPs) are still unknown because of the limitations of standards. Here, influent and effluent samples from 148 municipal WWTPs in China were collected. Ion mobility spectrometry was introduced into high-resolution mass spectrometry-based suspect screening methods to identify the target and suspect PFAS isomers. A total of 38 branched isomers of 14 typical PFASs were identified in wastewater samples. Linear PFASs had higher detection rates (22.3%-100%) than branched isomers (2.0%-98%). Compared to the influents, proportions of branched isomers of most PFASs (except for perfluoropentanoic acid and perfluorohexanoic acid) increased in the effluents. The conventional biological treatment processes (such as anaerobic-anoxic-aerobic and oxidation ditch treatments) had poor removal efficiency for linear PFASs (<21.4%) and branched isomers (<13.4%). No difference on removal efficiency among treatment processes was found. Furthermore, isomer composition in the WWTPs showed obvious differences between East China region and other regions, and the usage of short-chain PFASs (perfluorobutanesulfonic acid and perfluorohexanesulfonic acid) may be a key factor for driving this difference. This study sheds lights on the identification and characterization of PFAS isomers in WWTPs, which would be useful for development of monitoring and control strategies of PFASs.

How anaerobic sludge microbiome respond to different concentrations of nitrite, nitrate, and ammonium ions: a comparative analysis.

High concentrations of ammonium, nitrite, and nitrate always induce inhibition in anaerobic wastewater treatment. Due to the complexity and vulnerability of the microbial community (especially methanogens) in anaerobic sludge, little is understood about its underlying microbial mechanism under such inhibition. In this study, the shifts of microbial communities in anaerobic sludge under increasing levels of nitrite, nitrate, and ammonium ions were compared. Results show that although half maximal inhibitory concentrations (methanogenesis) were different for nitrite, nitrate, and ammonium ions with EC50 values of 12, 30, and 3000 mg N/L, respectively, bacteria genera Kosmotoga and Brooklawnia dominated in all of the three high-stress inhibitory systems. Network analysis and redundancy analysis (RDA) of the microbial community showed the treatments with nitrate and nitrite ions decreased the modularity of anaerobic microorganisms. RDA showed that specific methanogenic activity was positively related to coenzyme F420 under nitrite inhibition (rp = 0.833, p < 0.05) and closely correlated with viability under nitrate inhibition. Gram-positive and nonmotile Brooklawnia genus showed a negative correlation with physiological characteristics in the ammonia treatments, suggesting its high resistance to ammonia.