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Nitrification, a process carried out by aerobic microorganisms that oxidizes ammonia to nitrate via nitrite, is an indispensable step in wastewater nitrogen removal. To facilitate energy and carbon savings, applying low dissolved oxygen (DO) is suggested to shortcut the conventional biological nitrogen removal pathway, however, the impact of low DO on nitrifying communities within activated sludge is not fully understood. This study used genome-resolved metagenomics to compare nitrifying communities under extremely low- and high-DO. Two bioreactors were parallelly operated to perform nitrification and DO was respectively provided by limited gas-liquid mass transfer from the atmosphere (AN reactor, DO < 0.1 mg/L) and by sufficient aeration (AE reactor, DO > 5.0 mg/L). Low DO was thought to limit nitrifiers growth; however, we demonstrated that complete nitrification could still be achieved under the extremely low-DO conditions, but with no nitrite accumulation observed. Kinetic analysis showed that after long-term exposure to low DO, nitrifiers had a higher oxygen affinity constant and could maintain a relatively high nitrification rate, particularly at low levels of DO (<0.2 mg/L). Community-level gene analysis indicated that low DO promoted enrichment of nitrifiers (the genera Nitrosomonas and Nitrospira, increased by 2.3- to 4.3-fold), and also harbored with 2.3 to 5.3 times higher of nitrification functional genes. Moreover, 46 high-quality (>90 % completeness and <5 % contamination) with 3 most abundant medium-quality metagenome-assembled genomes (MAGs) were retrieved using binning methods. Genome-level phylogenetic analysis revealed the species succession within nitrifying populations. Surprisingly, compared to DO-rich conditions, low-DO conditions were found to efficiently suppressed the ordinary heterotrophic microorganisms (e.g., the families Anaerolineales, Phycisphaerales, and Chitinophagales), but selected for the specific candidate denitrifiers (within phylum Bacteroidota). This study provides new microbial insights to demonstrate that low-DO favors the enrichment of autotrophic nitrifiers over heterotrophs with species-level successions, which would facilitate the optimization of energy and carbon management in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

PFOA has garnered heightened scrutiny for its impact on denitrification, especially given its frequent detection in secondary effluent discharged from wastewater treatment plants. However, it is still unclear what potential risk PFOA release poses to a typical advanced treatment process, especially the sulfur-based autotrophic denitrification (SAD) process. In this study, different PFOA concentration were tested to explore their impact on denitrification kinetics and microbial dynamic responses of the SAD process. The results showed that an increase PFOA concentration from 0 to 1000 μg/L resulted in a decrease in nitrate removal rate from 9.52 to 7.73 mg-N/L·h. At the same time, it increased nitrite accumulation and N 2 O emission by 6.11 and 2.03 times, respectively. The inhibitory effect of PFOA on nitrate and nitrite reductase activity in the SAD process was linked to the observed fluctuations in nitrate and nitrite levels. It is noteworthy that nitrite reductase was more vulnerable to the influence of PFOA than nitrate reductase. Furthermore, PFOA showed a significant impact on gene expression and microbial community. Metabolic function prediction revealed a notable decrease in nitrogen metabolism and an increase in sulfur metabolism under PFOA exposure. This study highlights that PFOA has a considerable inhibitory effect on SAD performance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Fumarate and nitrate reduction regulatory protein (Fnr)-a global transcriptional regulator-can directly or indirectly regulate many genes in different metabolic pathways at the top of the bacterial transcription regulation network. The present study explored the regulatory mechanism of Fnr-mediated nitrite degradation in Lactobacillus plantarum WU14 through gene transcription and expression analysis of oxygen sensing and nir operon expression regulation by Fnr. The interaction and the mechanism of transcriptional regulation between Fnr and GlnR were also examined under nitrite stress. After Fnr and GlnR purification by glutathione S-transferase tags, they were successfully expressed in Escherichia coli by constructing an expression vector. The results of electrophoresis mobility shift assay and qRT-PCR indicated that Fnr specifically bound to the PglnR and Pnir promoters and regulated the expression of nitrite reductase (Nir) and GlnR. After 6-12 h of culture, the expressions of fnr and nir under anaerobic conditions were higher than under aerobic conditions; the expression of these two genes increased with sodium nitrite (NaNO 2 ) addition during aerobic culture. Overall, the present study indicated that Fnr not only directly participated in the expression of Nir and GlnR but also indirectly regulated the expression of Nir through GlnR regulation., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background/aim: Alkaline extracts of several plants which contain lignin degradation products have several unique biological activities. In order to search for new biological activities of alkaline extracts of pine seed shell (APs), their anti-ultraviolet C (UVC) and macrophage stimulation activity were investigated., Materials and Methods: Anti-UVC activity was determined by the ratio of the 50% cytotoxic concentration against human melanoma cell line COLO679 to the 50% UVC-protective concentration. Extracellularly secreted nitrite (NO 2 - ) by unstimulated and lipopolysaccharide (LPS)-stimulated mouse macrophage-like cells RAW264.7 was determined by Griess method., Results: APs showed significantly higher anti-UVC activity than previously reported hot-water extracts of medical herbs. Anti-UVC activity of AP and vanillic acid was maintained for much longer than that of sodium ascorbate and vanillin. APs enhanced the production of NO 2 - to the level induced by LPS. Simultaneous addition of AP and LPS did not further increase NO 2 - production, suggesting their mechanisms of action overlap., Conclusion: The present study suggests the possible application of APs as UVC protectors and immunopotentiators via macrophage activation., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Simultaneous nitritation and denitritation have the potential to significantly improve nitrogen removal in sewage treatment processes. However, their application in low-strength sewage treatment systems presents challenges. This study explored the impact of four solid carbon sources (SCSs) on N-removal via nitrite in a multi-cycle SBR with biocarriers. Results showed that both N-removal efficiencies and nitrite accumulation rates increased with higher COD/N ratios, indicating that high COD/N ratios can improve the competition between denitrifiers and nitrite-oxidizing bacteria for nitrite, leading to stable simultaneous nitritation and denitritation. The supplementation of SCSs further enhanced this high-efficiency N-removal process, with polybutylene succinate (PBS) and polycaprolactone (PCL) showing greater increases in N-removal via nitrite than poly-hydroxybutyrate (PHB) and poly-hydroxyalkanoate (PHA). Moreover, PBS showed the most significant increase in denitrification efficiency in anoxic conditions, while PHA was the most effective external SCS at a moderate level of dissolved oxygen. These findings suggest that the incorporation of external SCSs can facilitate the simultaneous nitritation and denitrification process in multi-cycle SBRs, underscoring the importance of selecting an appropriate SCS for optimizing nitrogen removal in sewage treatment projects., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Guangjing Xu reports financial support was provided by National Natural Science Foundation of China. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Phenol, quinoline, and pyridine, commonly found in industrial wastewater, disrupt the nitrification process, leading to nitrite accumulation. This study explores the potential mechanisms through which these biotoxic organic compounds affect nitrite accumulation, using metagenomic and molecular docking analyses. Despite increasing concentrations of these compounds from 40 to 160 mg/L, ammonia nitrogen removal was not hindered, and stable nitrite accumulation rates exceeding 90 % were maintained. Additionally, these compounds inhibited nitrite-oxidizing bacteria (NOB) and enriched ammonia-oxidizing bacteria (AOB) in situ. As the concentration of these compounds rose, protein (PN) and polysaccharide (PS) concentrations also increased, along with a higher PN/PS ratio. Metagenomic analysis further revealed an increase in hao relative abundance, while microbial community analysis showed increased Nitrosomonas abundance, which contributed to nitrite accumulation stability. Molecular docking indicated that these compounds have lower binding energy with hydroxylamine oxidoreductase (HAO) and nitrate reductase (NAR), theoretically supporting the observed sustained nitrite accumulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

To provide the necessary nitrite for the Anaerobic Ammonium Oxidation (ANAMMOX) process, the effect of nitrite accumulation in the partial sulfide autotrophic denitrification (PSAD) process was investigated using an SBR reactor. The results revealed that the effectiveness of nitrate removal was unsatisfactory when the S/N ratio (mol/mol) fell below 0.6. The optimal conditions for nitrate removal and nitrite accumulation were achieved within the S/N ratio range of 0.7-0.8, resulting in an average Nitrate Removal Efficiency (NRE) of 95.84%±4.89% and a Nitrite Accumulation Rate (NAR) of 75.31%±6.61%, respectively. It was observed that the nitrate reduction rate was three times faster than that of nitrite reduction during a typical cycle test. Furthermore, batch tests were conducted to assess the influence of pH and temperature conditions. In the pH tests, it became evident that the PSAD process performed more effectively in alkaline environment. The highest levels of nitrate removal and nitrite accumulation were achieved at an initial pH of 8.5, resulting in a NRE of 98.30%±1.93% and a NAR of 85.83%±0.47%, respectively. In the temperature tests, the most favourable outcomes for nitrate removal and nitrite accumulation were observed at 22±1 ℃, with a NRE of 100.00% and a NAR of 81.03%±1.64%, respectively. Moreover, a comparative analysis of 16S rRNA sequencing results between the raw sludge and the sulfide-enriched culture sludge sample showed that Proteobacteria (49.51%) remained the dominant phylum, with Thiobacillus (24.72%), Prosthecobacter (2.55%), Brevundimonas (2.31%) and Ignavibacterium (2.04%) emerging as the dominant genera, assuming the good nitrogen performance of the system.

Achieving mainstream short-cut nitrogen removal via nitrite has become a carbon and energy efficient way, but still remains challenging for low-strength municipal wastewaters. This study integrated sidestream enhanced biological phosphorus removal system in a pilot-scale adsorption/bio-oxidation (A-B) process (named A-B-S2EBPR system) and nitrite accumulation was successfully achieved for treating the municipal wastewater. Nitrite could accumulate to 5.5 ± 0.3 mg N/L in the intermittently aerated tanks of B-stage with the nitrite accumulation ratio (NAR) of 79.1 ± 6.5 %. The final effluent concentration and removal efficiency of total inorganic nitrogen (TIN) were 4.6 ± 1.8 mg N/L and 84.9 ± 5.6 %, respectively. In-situ process performance of nitrogen conversions, routine batch nitrification/denitrification activity tests and functional gene abundance of nitrifiers collectively suggested that the nitrite accumulation was mainly caused by partial denitrification rather than out-selection of nitrite oxidizing bacteria (NOB). Moreover, the single-cell Raman spectroscopy analysis first demonstrated that there was a specific microbial population that could utilize polyhydroxyalkanoates (PHA) as the potential internal carbon source during the partial denitrification process. The integration of S2EBPR brings unique features to the conventional A-B process, such as extended anaerobic retention time, lower oxidation-reduction potential (ORP), much higher and complex volatile fatty acids (VFAs) etc., which can largely reshape the microbial communities. The dominant genera were Acinetobacter and Comamonadaceae, which accounted for (17.8 ± 15.5)% and (6.7 ± 3.4)%, respectively, while the relative abundance of conventional nitrifiers was less than 0.2%. This study provides insights into phylogenetic and phenotypic shifts of microbial communities when incorporating S2EBPR into the sustainable A-B process to achieve mainstream short-cut nitrogen removal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Methane-dependent complete denitrification primarily involves nitrate reduction to nitrite by ANME-2d archaea and nitrite reduction to dinitrogen by Methylomirabilis bacteria. 'Candidatus Methylomirabilis sinica' integrates the divisional labor. Physiological traits of this bacterium potentially enable the simultaneous reduction of N 2 O and CH 4 emissions. This forum article explores these traits and possible microbial mechanisms for co-reduction, providing guidance for greenhouse gas management strategies., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Reducing nitrites tends to increase the accumulation of hazardous biogenic amines (BAs) in Chinese fermented sausages (CFSs). Gallic acid (GA) has emerged as a potential alternative to reduce nitrite usage and control BAs. This study explored how GA inhibits BAs and nitrosamines accumulation in reduced-nitrite CFSs. Results demonstrated that combining 0.05% (w/w) GA with reduced nitrite effectively curbed BAs and N-nitrosodimethylamine, decreasing total BA from 271.48 to 125.46 mg/kg. Fifty-one metabolites associated with the metabolism of BAs and N-nitrosodimethylamine were identified. GA boosted Lactococcus while reducing spoilage bacteria and Macrococcus. This dual regulation suppressed BAs and dimethylamine accumulation by regulating amino acids and trimethylamine pathways. Consequently, GA achieved an 89.86% reduction in N-nitrosodimethylamine by decreasing the key precursors like putrescine, dimethylamine, and nitrite. These findings offer new insights into utilizing GA and similar plant polyphenols to manage BAs and nitrosamines in meat products with reduced nitrite usage., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Strain JAM1 T and strain GP59 of the methylotrophic, bacterial species Methylophaga nitratireducenticrescens were isolated from a microbial community of the biofilm that developed in a fluidized-bed, methanol-fed, marine denitrification system. Despite of their common origin, both strains showed distinct physiological characters towards the dynamics of nitrate ( NO 3 - ) reduction. Strain JAM1 T can reduce NO 3 - to nitrite ( NO 2 - ) but not NO 2 - to nitric oxide (NO) as it lacks a NO-forming NO 2 - reductase. Strain GP59 on the other hand can carry the complete reduction of NO 3 - to N 2 . Strain GP59 cultured under anoxic conditions shows a 24-48h lag phase before NO 3 - reduction occurs. In strain JAM1 T cultures, NO 3 - reduction begins immediately with accumulation of NO 2 - . Furthermore, NO 3 - is reduced under oxic conditions in strain JAM1 T cultures, which does not appear in strain GP59 cultures. These distinct characters suggest differences in the regulation pathways impacting the expression of denitrification genes, and ultimately growth., Methods: Both strains were cultured under oxic conditions either with or without NO 3 - , or under anoxic conditions with NO 3 - . Transcript levels of selected denitrification genes ( nar1 and nar2 encoding NO 3 - reductases, nirK encoding NO 2 - reductase, narK12f encoding NO 3 - / NO 2 - transporter) and regulatory genes ( narXL and fnr ) were determined by quantitative reverse transcription polymerase chain reaction. We also derived the transcriptomes of these cultures and determined their relative gene expression profiles., Results: The transcript levels of nar1 were very low in strain GP59 cultured under oxic conditions without NO 3 - . These levels were 37 times higher in strain JAM1 T cultured under the same conditions, suggesting that Nar1 was expressed at sufficient levels in strain JAM1 T before the inoculation of the oxic and anoxic cultures to carry NO 3 - reduction with no lag phase. Transcriptomic analysis revealed that each strain had distinct relative gene expression profiles, and oxygen had high impact on these profiles. Among denitrification genes and regulatory genes, the nnrS3 gene encoding factor involved in NO-response function had its relative gene transcript levels 5 to 10 times higher in strain GP59 cultured under oxic conditions with NO 3 - than those in both strains cultured under oxic conditions without NO 3 - . Since NnrS senses NO, these results suggest that strain GP59 reduced NO 3 - to NO under oxic conditions, but because of the oxic environment, NO is oxidized back to NO 3 - by flavohemoproteins (NO dioxygenase; Hmp), explaining why NO 3 - reduction is not observed in strain GP59 cultured under oxic conditions., Conclusions: Understanding how these two strains manage the regulation of the denitrification pathway provided some clues on how they response to environmental changes in the original biofilm community, and, by extension, how this community adapts in providing efficient denitrifying activities., Competing Interests: The authors declare there are no competing interests., (©2024 Lestin and Villemur.)

Epilepsy, a widespread neural ailment considered by prolonged neuronal depolarization and repetitive discharge, has been linked to extreme stimulus of N-methyl-D-aspartate receptors (NMDARs). Despite the availability of approved anti-seizure medications (ASMs) in many developed nations, approximately 30% of epilepsy patients continue to experience drug-resistant seizures. Thus, a growing interest in discovering natural compounds as potential sources for new medications is growing. Sinapinic acid, a natural derivative of cinnamic acid found in food sources, is known for its neuroprotective properties. This study investigated how sinapinic acid interacts with NMDA receptors and its potential role in providing anticonvulsant effects. Male mice were randomly allocated into nine groups: a control group receiving normal saline (1 ml/kg), groups treated with sinapinic acid at doses of 1, 3, and 10 mg/kg, a group treated with diazepam at 10 mg/kg, a group treated with an NMDA agonist at 75 mg/kg, a group treated with an NMDA antagonist at 0.5 mg/kg, a group receiving the ineffective dose of sinapinic acid (1 mg/kg) along with the NMDA antagonist, and a group receiving the effective dose of sinapinic acid (10 mg/kg) along with the NMDA agonist. Sinapinic acid and other treatments were administered intraperitoneally 30 min prior to inducing seizures with PTZ injection. Seizure onset time was recorded following PTZ injection. Blood and brain samples were collected after anesthesia to determine serum and brain nitrite levels. Real-time PCR assessed NMDAR gene expression in the prefrontal cortex (PFC). Data were analyzed using Prism software. The time seizures began was notably extended in groups treated with sinapinic acid at doses of 3 and 10 mg/kg compared to those treated with saline (P < 0.05). Additionally, in the receiving group of an ineffective dose of sinapinic acid alongside ketamine, the beginning of seizure time was significantly prolonged compared to the group that received the ineffective dose of sinapinic acid alone (P < 0.05). Serum and prefrontal cortex (PFC) nitrite levels were significantly lower in mice treated with sinapinic acid at doses of 1, 3, and 10 mg/kg compared to the saline-treated group (P < 0.05). The gene expression of the NMDAR NR2B subunit in the PFC was decreased in groups treated with sinapinic acid at 1 and 10 mg/kg compared to the saline-treated group. Furthermore, co-administration of sinapinic acid (10 mg/kg) with NMDA resulted in significantly lower NR2A gene expression than the group treated with 10 mg/kg of sinapinic acid alone. Conversely, co-administration of ketamine with sinapinic acid (1 mg/kg) significantly increased NR2B subunit gene expression compared to the group treated with sinapinic acid at 1 mg/kg alone. Sinapinic acid showed anticonvulsant effects through reduced serum and PFC nitrite and modulation of glutamatergic signaling., (© 2024. The Author(s).)

Background: Many bacteria are capable of reducing selenium oxyanions, primarily selenite (SeO 3 2- ), in most cases forming selenium(0) nanostructures. The mechanisms of these transformations may vary for different bacterial species and have so far not yet been clarified in detail. Bacteria of the genus Azospirillum , including ubiquitous phytostimulating rhizobacteria, are widely studied and have potential for agricultural biotechnology and bioremediation of excessively seleniferous soils, as they are able to reduce selenite ions., Methods: Cultures of A.brasilense Sp7 and its derivatives (mutant strains) were grown on the modified liquid malate salt medium in the presence or absence of selenite. The following methods were used: spectrophotometric monitoring of bacterial growth; inhibition of glutathione (GSH) synthesis in bacteria by L-buthionine-sulfoximine (BSO); optical selenite and nitrite reduction assays; transmission electron microscopy of cells grown with and without BSO and/or selenite., Results: In a set of separate comparative studies of nitrite and selenite reduction by the wild-type strain A.brasilense Sp7 and its three specially selected derivatives (mutant strains) with different rates of nitrite reduction, a direct correlation was found between their nitrite and selenite reduction rates for all the strains used in the study. Moreover, for BSO it has been shown that its presence does not block selenite reduction in A.brasilense Sp7., Conclusions: Evidence has been presented for the first time for bacteria of the genus Azospirillum that the denitrification pathway known to be inherent in these bacteria, including nitrite reductase, is likely to be involved in selenite reduction. The results using BSO also imply that detoxification of selenite through the GSH redox system (which is commonly considered as the primary mechanism of selenite reduction in many bacteria) does not play a significant role in A.brasilense . The acquired knowledge on the mechanisms underlying biogenic transformations of inorganic selenium in A.brasilense is a step forward both in understanding the biogeochemical selenium cycle and to a variety of potential nano- and biotechnological applications., (© 2024 The Author(s). Published by IMR Press.)

Introduction: Human aortic tissues in vitro are tools to clarify the pathophysiological mechanisms of the cardiovascular system, cell culture, and transplants. Therefore, this study aims to analyze and compare the preservation of human aneurysmatic aortic tissues in three different solutions., Methods: Six human abdominal aortic aneurysms were obtained from patients after surgical ablation. The aorta samples were incubated in different solutions - 0.9% normal physiological saline solution, Ringer's lactate solution, and histidine-tryptophan-ketoglutarate solution (Custodiol®). Segments were collected at 0, 6, 24, and 48 hours. Creatine kinase and nitrate/nitrite were quantified for each incubation time. The tissue's alpha-smooth muscle actin was analyzed by immunofluorescence., Results: There was a significant increase in creatine kinase formation in the normal saline group at 0 and 48 hours and in the Ringer's lactate group at 0 and 48 hours (P=0.018 and P=0.028). The lower levels of creatine kinase and nitrate/nitrite and the aortic tissues' morphological integrity show that histidine-tryptophan-ketoglutarate has better tissue protection. These data suggest that histidine-tryptophan-ketoglutarate induces a protective effect on smooth muscle cells, with less tissue depletion in the aortic aneurysm., Conclusion: This study compared three preservation solutions with the potential for human abdominal aortic aneurysm tissue preservation. The histidine-tryptophan-ketoglutarate solution reduced tissue injury and improved tissue preservation in human abdominal aortic aneurysm tissue samples.

Anammox bacteria are obligate anaerobic bacteria that exist widely in nature with sufficient amounts of dissolved oxygen. However, whether anammox bacteria can grow under aerobic conditions remains unclear. In this study, we found that the production of nitrate in the anammox system under aerobic conditions was significantly higher than that under anaerobic conditions without total nitrogen loss. Anammox bacteria can grow by oxidizing nitrite and dehydrogenating hydrazine to produce electrons for carbon fixation. The hydrazine dehydrogenase in anammox bacteria was inhibited under aerobic conditions, and the nitrite oxidoreductase transcription expression of anammox bacteria increased by 2.7 times compared to that under anaerobic conditions, which was the main way for anammox bacteria perform carbon fixation. DNA-stable isotope probing with 13 C bicarbonate found the existence of anammox bacteria with 13 C isotopes in aerobic cultivation, further proving that anammox bacteria can grow under aerobic condition. More than half of the pathways in glycolysis, the Wood-Ljungdahl pathway, and the tricarboxylic acid cycle were upregulated in anammox bacteria in aerobic condition. Large amounts of bacterioferritins are the important antioxidative enzymes in anammox bacteria in the aerobic environment, which contributes to their stronger oxygen adaptation than other anaerobes. This study expands our understanding of the growth mechanism of anammox bacteria as well as the oxygen adaptation strategies of obligate anaerobic bacteria.

Partial nitrification (PN) is crucial for anaerobic ammonium oxidation (ANAMMOX), but faces challenges such as high energy demands and process control. Recent research has highlighted additives like magnetite as potential alternatives to conventional electron acceptors (O₂ and NO₂ - ) for enhancing ammonium (NH 4 + ) oxidation with lower energy consumption. This study investigated the effect of adding 50 mg/L of magnetite to ANAMMOX reactors, resulting in improved nitrogen (N) removal efficiency. The magnetite-added ANAMMOX (M-ANA) reactor yielded N removal efficiencies of 71 %, 66 %, and 57 % for NH 4 + :NO 2 - molar ratios of 1:1.3, 1:0.8, and 1:0.5, respectively. The M-ANA reactor operated under a 0.5 mol lower NO 2 - concentration achieved similar performance to the control ANAMMOX (C-ANA) reactor operated with a theoretical amount of NO 2 - . Moreover, the M-ANA reactor showed the potential to remove NH 4 + by 56 % without any NO 2 - supplementation. Metagenomic analysis showed that the addition of magnetite significantly improved the relative abundance of microorganisms involved in the FEAMMOX reaction, such as Fimbriimonas ginsengisoli and Pseudomonas stutzeri. It also facilitated positive mutualism between ANAMMOX and FEAMMOX reactions. In addition, M-ANA granules exhibited a dense and compact structure compared with C-ANA, and the presence of magnetite facilitated the formation of resilient granules. Notably, the useful protein (Heme C) concentration and specific microbial activity in the M-ANA reactor were 1.3 and 2.2 times higher than those in the C-ANA reactor. Overall, the results demonstrate that an appropriate amount of magnetite can enhance the N removal efficiency while reducing the energy input requirements and associated carbon emissions. These findings can guide the future development of carbon- and energy-neutral N removal processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this study., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The intensive use of various antibiotics for clinical and agricultural purposes has resulted in their widespread use in wastewater treatment plants. However, little research has been conducted on the effects of antibiotics on nitrite accumulation, antibiotic degradation pathways, or the microbial community structure in nitrification systems. In this study, a laboratory-scale sequencing batch reactor was used to treat wastewater containing cefalexin (CFX) at different doses (5, 10, 15, and 20 mg/L). The results showed that the nitrification performance was gradually inhibited with increasing CFX concentration. Ammonia-oxidizing bacteria (AOB) are more tolerant to CFX than nitrite-oxidizing bacteria (NOB). Under 15 mg/L of CFX, NOB were completely suppressed, whereas AOB were partially inhibited, as evidenced by an ammonium removal efficiency of 60 % and a 90 % of nitrite accumulation ratio. The partial nitritation was achieved. CFX can be degraded into 2-hydroxy-3phenylpyrazine and cyclohexane through bacterial co-metabolism, and CFX degradation gradually diminishes with decreasing nitrification performance. The abundance of Nitrospira gradually decreased with increasing CFX concentration. Ferruginibacter, Hydrogenophaga, Thauera, and Pseudoxanthomonas were detected at relative abundances of 13.2 %, 0.4 %, 0.9 %, and 1.3 %, respectively, indicating their potential roles in antibiotic degradation. These findings provide insight into the interactions between antibiotics and microbial communities, which are beneficial for a better understanding of antibiotic degradation in nitrification systems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Anaerobic ammonium oxidation (anammox)-the biological process that activates ammonium with nitrite-is responsible for a significant fraction of N 2 production in marine environments. Despite decades of biochemical research, however, no synthetic models capable of anammox have been identified. Here we report that a copper sulfide mineral replicates the entire biological anammox pathway catalysed by three metalloenzymes. We identified a copper-nitrosonium {CuNO} 10 complex, formed by nitrite reduction, as the oxidant for ammonium oxidation that leads to heterolytic N-N bond formation from nitrite and ammonium. Similar to the biological process, N 2 production was mediated by the highly reactive intermediate hydrazine, one of the most potent reductants in nature. We also found another pathway involving N-N bond heterocoupling for the formation of hybrid N 2 O, a potent greenhouse gas with a unique isotope composition. Our study represents a rare example of non-enzymatic anammox reaction that interconnects six redox states in the abiotic nitrogen cycle., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

The anammox reaction simultaneously utilizes ammonia and nitrite as substrates; however, high nitrite concentrations act as strong inhibitors of the reaction. In this study, inhibition by NO 2 - and free nitrous acid (FNA) was separately evaluated in continuous feeding tests using different biomass carriers. The influent NO 2 - concentration was increased under pH 7.6, where FNA is less likely to affect anammox activity. A continuous test using polyethylene glycol (PEG) gel carriers containing immobilized anammox bacteria showed that the inhibition ratio was 13% when the NO 2 - -N concentration in the reactor was 350 mg L -1 (FNA ≤0.06 mg L -1 ). The relationship between NO 2 - concentration in the reactor and inhibition ratio increased linearly. Evaluation of the inhibitory effect of FNA by increasing the influent NO 2 - concentration at pH 6.4, where FNA is easily formed, demonstrated that the relationship between FNA and inhibition ratio could be fitted to a sigmoid curve, and the 50% inhibitory concentration (IC 50 ) of FNA was 0.88 mg L -1 . A similar test performed using polyvinyl alcohol carriers containing anammox bacteria on their surface showed the same trend as the PEG gel carriers, with the IC 50 for FNA at 0.70 mg L -1 . These results indicate that the inhibitory effect of FNA on anammox activity was greater than that of NO 2 - . The evaluation of these two factors helped identify important operational indicators of the stable application of anammox processes., (Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Nitrite is a common environmental pollutant in intensive aquaculture systems. In this study, physiological and transcriptomic analyses were performed to investigate nitrite stress responses in the swimming crab Portunus trituberculatus, an important aquaculture species in China. The results revealed that nitrite can affect neurotransmitter signaling via the expression of neurotransmitter receptors such as octopamine receptor (OAR) and 5-hydroxytryptamine receptor (5-HTR), and depress ecdysteroid signaling by downregulating ecdysteroid receptor (EcR) as well as its downstream transcription factors in hepatopancreas. In addition, nitrite suppressed the expression of hemocyanins, the oxygen-transporting protein, which at least partly contributed to tissue hypoxia, resulting in a switchover of energy metabolism from aerobic to anaerobic pathway. To meet the energy demand, glycogens and lipids were mobilized and transported to the hemolymph, and the catabolism of amino acids and fatty acids was enhanced to provide energy for hepatopancreas. β-oxidation of fatty acids, the major process by which fatty acids are oxidized to generate energy, seems to occur mainly not in mitochondria but in peroxisomes. Although the cellular protective mechanisms, including antioxidant defense, heat shock response (HSR), unfolded protein response (UPR), and autophagy, were activated, nitrite-induced cellular stress overwhelmed the repairing capacity and caused significant increase in the levels of apoptosis. These results indicated that nitrite stress influences neurotransmitter and endocrine signaling, disturbs energy metabolism, damages cellular components, and induces apoptosis in P. trituberculatus. The findings of this study provide new insights into nitrite stress response in the swimming crab and provide valuable information for aquaculture management of this species., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with worldwide increasing incidence. Maternal separation (MS) stress at the beginning of life with its own neuroendocrine changes can provide the basis for development of ASD. Rosmarinic acid (RA) is a phenolic compound with a protective effect in neurodegenerative diseases. The aim of this study was to determine the effect of RA on autistic-like behaviors in maternally separated mice focusing on its possible effects on neuroimmune response and nitrite levels in the hippocampus. In this study, 40 mice were randomly divided into five groups of control (received normal saline (1 ml/kg)) and MS that were treated with normal saline (1 ml/kg) or doses of 1, 2, and 4 mg/kg RA, respectively, for 14 days. Three-chamber sociability, shuttle box, and marble burying tests were used to investigate autistic-like behaviors. Nitrite level and gene expression of inflammatory cytokines including TNF-α, IL-1β, TLR4, and iNOS were assessed in the hippocampus. The results showed that RA significantly increased the social preference and social novelty indexes, as well as attenuated impaired passive avoidance memory and the occurrence of repetitive and obsessive behaviors in the MS mice. RA reduced the nitrite level and gene expression of inflammatory cytokines in the hippocampus. RA, probably via attenuation of the nitrite level as well as of the neuroimmune response in the hippocampus, mitigated autistic-like behaviors in maternally separated mice., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study explored a novel economical and efficient process for treating actual low-ammonia nitrogen electroplating tail wastewater. A pilot scale system of denitrification-partial nitrification/anaerobic ammonium oxidation (DN-PN/A) was constructed and operated for 190 days. The partial nitrification (PN) reactor, filled with zeolite, increased free ammonia concentration beyond the nitrite oxidizing bacteria threshold and successfully supplied NO 2 - -N, with nitrite accumulation rate exceeding 90 %. Over 109 days, the total nitrogen removal rate achieved was 80.2 ± 7.41 %, and the chemical oxygen demand removal rate reached 79.68 ± 9.53 %. The dominant functional bacteria were Nitrosomonas (5.45 %) and Candidatus Anammoxoglobus (28.84 %) in PN reactor and anaerobic ammonium oxidation (Anammox) reactor. This process, characterized by rapid start-up, strong shock resistance, and low cost, alleviates the pressure of ammonium pollution control, promotes the sustainable development of the electroplating industry and has the potential for application in the treatment of other industrial wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

High Pressure Hydrogenotrophic Denitrification (HPHD) provided a promising alternative for efficient and clean nitrate removal. In particular, the denitrification rates at low temperature could be compensated by elevated H 2 partial pressure. However, nitrite reduction was strongly inhibited while nitrate reduction was barely affected at low temperature. In this study, the nitrate reduction gradually recovered under long-term low temperature stress, while nitrite accumulation increased from 0.1 to 41.0 mg N/L. The activities of the electron transport system (ETS), nitrate reductase (NAR), and nitrite reductase (NIR) decreased by 45.8 %, 27.3 %, and 39.3 %, respectively, as the temperature dropped from 30 °C to 15 °C. Real time quantitative PCR analysis revealed that the denitrifying gene expression rather than gene abundance regulated nitrogen biotransformation. The substantial nitrite accumulation was attributed to the significant up-regulation by 54.7 % of narG gene expression and down-regulation by 73.7 % of nirS gene expression in hydrogenotrophic denitrifiers. In addition, the nirS-gene-bearing denitrifiers were more sensitive to low temperature compared to those bearing nirK gene. The dominant populations shifted from the genera Paracoccus to Hydrogenophaga under long-term low temperature stress. Overall, this study revealed the microbial mechanism of high nitrite accumulation in hydrogenotrophic denitrification at low temperature., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Humic acid (HA), a common natural organic matter, could affect conventional anoxic denitrification. Aim of this study was to investigate effect of HA on the process of aerobic denitrification in Achromobacter sp. GAD-3, an aerobic denitrifying strain. The findings demonstrated that an increase in HA concentrations (≥5 mg L -1 ) promoted the aerobic denitrification process (excluding N 2 O reduction), manifesting as higher rates of nitrate removal (6.67-11.1 mg L -1  h -1 ) and lower levels of nitrite accumulation (30.2-20.7 mg L -1 ). This was attributed to the increased electron transfer activities and denitrifying reductase activities (including NAR, NIR and NOR) facilitated by HA. Accordingly, the expression of denitrification genes such as napA, cnorB, and nirS was enhanced by HA. Nonetheless, the nosZ gene and N 2 OR activity underwent suppression by HA, which was accountable for N 2 O emission. It is crucial to understand the HA mechanism towards aerobic denitrifiers for wastewater treatment plants to enhance nitrogen removal., (Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Introduction: Viral infections cause oxygen deprivation, leading to hypoxia or anoxia in certain tissues. The limitation of mitochondrial respiration is one of the major events during hypoxia that induces alternative metabolic activities and increased levels of certain biomolecules such as nitric oxide (NO) metabolites. In this study, we aimed to investigate the role of NO metabolites and hypoxia in HPV infection., Materials and Methods: We included 36 patients with palmoplantar warts and 36 healthy subjects and performed serum determinations of NO metabolites (direct nitrite, total nitrite, nitrate, and 3-nitrotyrosine) and HIF1α, a marker of hypoxia., Results: We found elevated serum levels in NO metabolites and HIF1α, and decreased direct nitrite/nitrate ratios in patients with warts versus controls. Additionally, we identified statistically significant positive correlations between NO metabolites and HIF1α levels, except for 3-nitrotyrosine., Conclusions: Our findings show that HPV infection causes hypoxia and alterations in NO metabolism and suggest a link between wart development and cellular stress. Our research could provide new insights for a comprehensive understanding of the pathogenesis of cutaneous HPV infections.

How to intensify the ammonia oxidation rate (AOR) is still a bottleneck impeding the technology development for the innovative acidic partial nitritation because the eosinophilic ammonia-oxidizing bacteria (AOB), such as Nitrosoglobus or Nitrosospira, were inhibited by the high-level free nitrous acid (FNA) accumulation in acidic environments. In this study, an innovative approach of dynamic acidic pH regulation control strategy was proposed to realize high-rate acidic partial nitritation driven by common AOB genus Nitrosomonas. The acidic partial nitrification process was carried out in a laboratory-scale sequencing batch moving bed biofilm reactor (SBMBBR) for long-term (700 days) to track the effect of dynamic acidic pH on nitrifying bacterial activity. The results indicated that the influent NH 4 + -N concentration was about 100 mg/L, the nitrite accumulation ratio was exceeding 90%, and the maximum AOR can reach 14.5 ± 2.6 mg N L -1 h -1 . Although the half-saturation inhibition constant of NOB (K I&#95;FNA(AOB) ) reached 0.37 ± 0.10 mg HNO 2 N/L and showed extreme adaptability in FNA, the inactivation effect of FNA (6.1 mg HNO 2 N/L) for NOB was much greater than that of AOB, with inactivation rates of 0.61 ± 0.08 h -1 and 0.06 ± 0.01 h -1 , respectively. The effluent pH was gradually reduced to 4.5 by ammonia oxidation process and the periodic FNA concentration reached 6.5 mg HNO 2 N/L to inactivate nitrite-oxidizing bacteria (NOB) without negatively affecting Nitrosomonas during long-term operation. This result provides new insights for the future implementation of high-rate stabilized acidic partial nitritation by Nitrosomonas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Global urbanization requires more stable and sustainable wastewater treatment to reduce the burden on the water environment. To address the problem of substrate inhibition of microorganisms during wastewater treatment, which leads to unstable wastewater discharge, this study proposes an approach to enhance the tolerance of bacterial community by artificially setting up a non-lethal high substrate environment. And the feasibility of this approach was explored by taking the inhibition of anammox process by nitrite as an example. It was shown that the non-lethal high substrate environment could enhance the nitrite tolerance of anammox bacterial community, as the specific anammox activity increasing up to 24.71 times at high nitrite concentrations. Moreover, the system composed of anammox bacterial community with high nitrite tolerance also showed greater resistance (two-fold) in response to nitrite shock. The antifragility of the system was enhanced without affecting the operation of the main reactor, and the non-lethal high nitrite environment changed the dominant anammox genera to Candidatus Jettenia. This approach to enhance tolerance of bacterial community in a non-lethal high substrate environment not only allows the anammox system to operate stably, but also promises to be a potential strategy for achieving stable biological wastewater treatment processes to comply with standards., (© 2024. The Author(s).)

The Asian clam, Corbicula fluminea, commonly used in engineered wetlands receiving tailwater, affects nitrogen compound transformation in water. This study investigates how a commonly observed antibiotic in tailwater, norfloxacin, impact nitrogen compound transformation in tailwater containing C. fluminea. The clam was exposed to artificial tailwater with norfloxacin (0, 0.2, 20, and 2000 μg/L) for 15 days. Water properties, C. fluminea ecotoxicity responses, microorganism composition and nitrification- or denitrification-related enzyme activities were measured. Results revealed norfloxacin-induced increases and reductions in tailwater NH 4 + and NO 2 - concentrations, respectively, along with antioxidant system inhibition, organ histopathological damage and disruption of water filtering and digestion system. Microorganism composition, especially biodiversity indices, varied with medium (clam organs and exposure water) and norfloxacin concentrations. Norfloxacin reduced NO 2 - content by lowering the ratio between microbial nitrifying enzyme (decreased hydroxylamine oxidoreductase and nitrite oxidoreductase activity) and denitrifying enzyme (increased nitrate reductase and nitrite reductase activity) in tailwater. Elevated NH 4 + content resulted from upregulated ammonification and inhibited nitrification of microorganisms in tailwater, as well as increased ammonia emission from C. fluminea due to organ damage and metabolic disruption of the digestion system. Overall, this study offers insights into using benthic organisms to treat tailwater with antibiotic residues, especially regarding nitrogen treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Polymesoda erosa is a mangrove clam known for its water filtration ability. This clam was investigated for its bioremediation potential and growth in synthetic wastewater during 40 days of incubation. Variation in the nutrient composition of water, biochemical composition of the clams, and metagenomic analysis of the microorganisms associated with clam tissue were carried out. Significant differences in the concentration of ammonia (p ≤ 0.01), nitrite (p ≤ 0.001), and nitrate (p ≤ 0.05) in the wastewater were observed between day 0 and day 40. A reduction of approximately 89% in ammonia concentration at the end of the experiment was recorded indicating nitrification activity. However, biochemical parameters showed negligible differences before and after the incubation experiment. Thus suggesting that the chemosynthetic-based nutrition aids in the survival of the clam as no organic matter was added to the medium. The substantial decline in levels of ammonia in the presence of clams as compared to its absence suggests the significant role of clams in improving the water quality. Furthermore, the metagenomic analysis of the gill tissue of P. erosa revealed ~ 50% of the microbial population to consist of nitrifiers. The study highlights the contribution by the nitrifers associated with the clams not only to its growth and resilience but also to bioremediation., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Two obligately acidophilic, mesophilic and aerobic soil ammonia-oxidising archaea were isolated from a pH 4.5 arable sandy loam (UK) and pH 4.7 acidic sulphate paddy soil (PR China) and designated strains Nd1 T and Nd2 T , respectively. The strains shared more than 99 % 16S rRNA gene sequence identity and their genomes were both less than 2 Mb in length, sharing 79 % average nucleotide identity, 81 % average amino acid identity and a DNA G+C content of approximately 37 mol%. Both strains were chemolithotrophs that fixed carbon dioxide and gained energy by oxidising ammonia to nitrite, with no evidence of mixotrophic growth. Neither strain was capable of using urea as a source of ammonia. Both strains were non-motile in culture, although Nd1 T does possess genes encoding flagella components and therefore may be motile under certain conditions. Cells of Nd1 T were small angular rods 0.5-1 µm in length and grew at pH 4.2-5.6 and at 20-30 °C. Cells of Nd1 T were small angular rods 0.5-1 µm in length and grew at pH 4.0-6.1 and at 20-42 °C. Nd1 T and Nd2 T are distinct with respect to genomic and physiological features and are assigned as the type strains for the species Nitrosotalea devaniterrae sp. nov. (type strain, Nd1 T =NCIMB 15248 T =DSM 110862 T ) and Nitrosotalea sinensis sp. nov. (type strain, Nd2 T =NCIMB 15249 T =DSM 110863 T ), respectively, within the genus Nitrosotalea gen. nov. The family Nitrosotaleaceae fam. nov. and order Nitrosotaleales ord. nov. are also proposed officially.

Anammox bacteria (AnAOB) can be easily enriched under high temperatures and high substrate concentrations, while the application of the mainstream anammox process in low substrate municipal sewage is still relatively uncommon. Therefore, this study investigated the enrichment of AnAOB under conditions of low ammonia nitrogen and nitrite concentration at 25 °C. Results showed that using inoculated aerobic sludge, four ASBRs (R1, R2, R3 and R4) were successfully initiated with different influent substrate (NO 2 - -N/NH 4 + -N) ratios of 1.2, 1.32, 1.4 and 1.5, respectively, with reactor start-up times were 162, 150, 120 and 134 days, respectively. The values of ΔNO 2 - -N/ΔNH 4 + -N in reactors were stable at 1.17, 1.32, 1.43 and 1.53 respectively. The increase in influent substrate ratios resulted in improved TN removal rates and accelerated consumption of chemical oxygen demand (COD) during the initial start-up stage. The maximum TN removal rates achieved in the four reactors were 76.09%, 79.24%, 82.82% and 82.63%, respectively. The color of sludge gradually changes from yellowish-brown to reddish-brown. Furthermore, the surface of sludge exhibited a porous mineral structure, with crater-like cavities. The dominant anammox species in the system was identified as Candida Brocadia (3.04%). According to qPCR, the abundance of hzsB in the system is 1.65 × 10 12 copies/g VSS, confirming the effective enrichment of AnAOB., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

This study elucidated the influence on a partial denitrification (PD) system under 0-1 mg/L sulfamethoxazole (SMX) stress in a sequencing batch reactor. The results showed that the nitrite accumulation rate (NAR) significantly (P ≤ 0.01) decreased from 68.68 ± 9.00% to 49.05 ± 11.68%, while the total nitrogen removal efficiency significantly (P ≤ 0.001) increased from 23.19 ± 4.42% to 31.36 ± 2.73% in presence of SMX. The results indicated that SMX exposure switched the PD process to complete denitrification through the deterioration of the nitrite accumulation and the promotion of further nitrite reduction. The SMX removal loading rate increased from 0.21 ± 0.04 to 5.03 ± 0.77 mg-SMX/(g-MLVSS·d) with the extended reactor operation under SMX stress. Low SMX concentration exposure increased extracellular polymeric substances (EPS) content from 107.69 ± 20.78 mg/g-MLVSS (0.05 mg-SMX/L) to 123.64 ± 9.66 mg/g-MLVSS (0.5 mg-SMX/L), while EPS secretion was inhibited under high SMX concentration exposure (i.e., 1 mg-SMX/L). Moreover, SMX exposure promoted the synthesis of aromatic protein-like compounds and changed the functional groups as revealed by EEM and FTIR analysis. Additionally, SMX exposure significantly shifted the microbial community structures at both phylum and genus levels. Particularly, the abundance of Thauera, i.e., functional bacteria related to PD, considerably decreased from 41.69% to 11.62% after SMX exposure, whereas the abundances of Denitratisoma and SM1A02 significantly rose under different SMX concentrations. These outcomes hinted that the addition of SMX resulted in the shifting of partial denitrification to complete denitrification., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Nitrate may act as a regulator of • NO bioavailability via sequential reduction along the nitrate-nitrite-NO pathway with widespread health benefits, including a eubiotic effect on the oral and gut microbiota. Here, we discuss the molecular mechanisms of microbiota-host communication through redox pathways, via the production of • NO and oxidants by the family of NADPH oxidases, namely hydrogen peroxide (via Duox2), superoxide radical (via Nox1 and Nox2) and peroxynitrite, which leads to downstream activation of stress responses (Nrf2 and NFkB pathways) in the host mucosa. The activation of Nox2 by microbial metabolites is also discussed. Finally, we propose a new perspective in which both oral and gut microbiota communicate through redox pathways, with nitrate as the pivot linking both ecosystems., (© 2024 Federation of European Biochemical Societies.)

Ischemic stroke occurs due a blockage in the blood flow to the brain, leading to damage to the nervous system. The prevalent morbidities resulting from stroke include post-stroke infection, as sepsis. Additionally, oxidative stress is recognized for inducing functional deficits in peripheral organs during sepsis. Therefore, sex differences in stroke exist and we aimed to investigate the peripheral oxidative stress caused by sepsis after stroke in male and female rats. Wistar rats (male and female) were divided into sham+sham, middle cerebral artery occlusion (MCAO) + sham, sham+ cecal ligation and perforation (CLP) and MCAO+CLP groups to males and female rats. Animals were subjected to MCAO or sham and after 7 days, were subjected to sepsis by CLP or sham. After 24 h, serum, total brain, lung, liver, heart, and spleen were collected. Brain edema, myeloperoxidase (MPO) activity, nitrite/nitrate (N/N) concentration, oxidative damage to lipids and proteins, and catalase activity were evaluated. Brain edema was observed only in male rats in MCAO+CLP group compared to MCAO+sham. Regarding MPO activity, an increase was verified in male in different organs and serum in MCAO+CLP group. For N/N levels, the increase was more pronounced in females submitted to MCAO+CLP. In general, to oxidative stress, an increase was only observed in animals exposed to MCAO+CLP, or with a greater increase in this group compared to the others. The findings provided the first indication that animals exposed to MCAO exhibit a heightened vulnerability to the harmful impacts of sepsis, as evidenced by brain edema and peripheral oxidative stress, and this susceptibility is dependent of sex., Competing Interests: Declaration of competing interest None of the authors or funding sources has conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Nitrospira is a common genus of nitrite-oxidising bacteria (NOB) found in wastewater treatment plants (WWTPs). To identify the key factors influencing the composition of NOB communities, research was conducted using both sequencing batch reactor (SBR) and continuous flow reactor under different conditions. High-throughput 16S rRNA gene sequencing revealed that Nitrospira (18.79% in R1 and 25.77% in R3) was the dominant NOB under low dissolved oxygen (DO) and low nitrite ( NO 2 - -N) concentrations, while Nitrobacter (21.26% in R2) was the dominant NOB under high DO and high NO 2 - -N concentrations. Flocculent and granule sludge were cultivated with Nitrospira as the dominant genus. Compared to Nitrospira flocculent sludge, Nitrospira granule sludge had higher inhibition threshold concentrations for free ammonia (FA) and free nitrous acid (FNA). It was more likely to resist adverse environmental disturbances. Furthermore, the effects of environmental factors such as temperature, pH, and DO on the activity of Nitrospira granular sludge were also studied. The results showed that the optimum temperature and pH for Nitrospira granular sludge were 36°C and 7.0, respectively. Additionally, Nitrospira granular sludge showed a higher dissolved oxygen half-saturation constant ( K o) of 3.67 ± 0.71 mg/L due to its morphological characteristics. However, the majority of WWTPs conditions do not meet the conditions for the Nitrospira granular sludge. Thus, it can be speculated that future development of aerobic partial nitrification granular sludge may automatically eliminate the influence of Nitrospira . This study provides a theoretical basis for a deeper understanding of Nitrospira and the development of future water treatment processes.

This study proposes an innovative strategy for achieving PN in synthetic domestic wastewater by side-stream sludge treatment using sulfide as the sole control factor. By conducting controllable batch experiments and response surface analysis, optimal sulfide treatment conditions were firstly determined as 90 mg/L of sulfide, 7.5 of pH, 100 rpm of rotation and 12 h of treatment time. After treatment, half of ammonia oxidizing bacteria (AOB) activity remained, but nitrite oxidizing bacteria (NOB) activity was barely detected. Nitrite accumulation rate of long-term running PN steadily reached 83.9 % with 99.1 % of ammonia removal efficiency. Sulfide treatment increased community diversity and facilitated stability of microbiota functioning with PN phenotype, which might be sustained by the positive correlation between ammonia oxidation gene (amoA) and sulfur oxidation gene (soxB). Correspondingly, the network analysis identified the keystone microbial taxa of persistent PN microbiota as Nitrosomonas, Thauera, Truepera, Defluviimonas and Sulitalea in the later stage of long-term reactor., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the impact of varying total ammonia nitrogen (TAN) feed levels along with water temperature decreases on the performance of nitrifying moving bed biofilm reactor (MBBR) at 1 °C and its recovery at 3 °C. Five MBBR reactors were operated with different TAN concentrations as water temperature decreased from 20 to 3 °C: reactor R1 at 30 mg N/L, reactor R2 at 20 mg N/L, reactor R3 at 15 mg N/L, reactor R4 at 10 mg N/L and reactor R5 at 0 mg N/L. The corresponding biofilm characteristics were also analyzed to understand further nitrifying MBBR under different TAN feeding scenarios. The findings revealed that the higher TAN levels were before reaching 1 °C, the better nitrification performance and the more biomass grew. However, the highest TAN concentration (30 mg N/L) might negatively affect the nitrification performance, the activity of nitrifiers, and the growth of biofilms at 1 °C because of the toxic effects of un-ionized or free ammonia (FA). It was observed that the activities of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were affected by FA concentrations ranging from 0.2 to 0.7 mg N/L at 1 °C, but they could gradually be adapted to such inhibitory environment, with NOB recovering more quickly and robustly than AOB. The study identified 20 mg N/L (67 % of maximum influent TAN at 1 °C in R2 as the optimal TAN feeding concentration, achieving over 90 % TAN removal and a surface area removal rate (SARR) of 0.78 ± 0.02 g N/m 2 ·d at 1 °C. Meanwhile, R2 also exhibited the highest biofilm mass, with total solids at 13.3 mg/carrier and volatile solids at 11.3 mg/carrier. As TAN was removed, nitrite accumulation was observed at 1 °C, and higher influent TAN concentrations prior to 1 °C appeared to delay the accumulation. When water temperature increased from 1 °C to 3 °C, nitrification performance improved significantly in all reactors without nitrite accumulation, and the higher TAN feeding in the previous stage led to faster recovery. Compared with 20 °C, biofilm became thinner and denser at 1 °C and 3 °C. Furthermore, this study revealed significant shifts in microbial community composition and nitrifier abundances in response to changes in water temperature and influent TAN levels. The dominant nitrifiers were identified as Nitrosomonadaceae (AOB) and Nitrospiraceae (NOB). At 1 °C, the nitrifier abundances were significantly correlated with SARRs, FA, and biofilm density. R2, which exhibited the best nitrification performance, maintained higher nitrifier abundances at 1 °C., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A novel A. pittii J08 with heterotrophic nitrification and aerobic denitrification (HN-AD) isolated from pond sediments could rapidly degrade inorganic nitrogen (N) and total nitrogen (TN-N) with ammonium (NH 4 + -N) preference. N degradation rate of NH 4 + -N, nitrite (NO 2 - -N) and nitrate (NO 3 - -N) were 3.9 mgL -1 h -1 , 3.0 mgL -1 h -1 and 2.7 mgL -1 h -1 , respectively. In addition, strain J08 could effectively utilize most of detected low-molecular-weight carbon (LMWC) sources to degrade inorganic N with a wide adaptability to various culture conditions. Whole genome sequencing (WGS) analysis revealed that assembled genome of stain J08 possessed the crucial genes involved in dissimilatory/assimilatory NO 3 - -N reduction and NH 4 + -N assimilation. These results indicated that strain J08 could be applied to wastewater treatment in aquaculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Rare earth elements result in substantial tailings wastewater with high ammonium and nitrate during extraction. In this study, a temperature-resilient Anammox process was employed for efficient treatment of rare earth element tailings wastewater through implementing synergistic nitrite supply by partial nitritation (PN) and partial denitrification (PD). Enhancing temperature resilience of Anammox process relies on dynamic management of DO and COD inputs to shift the dominant nitrite supplier from PN to PD, stable PD (NAR ≥ 90 %) can boost nitrogen removal by Anammox to 97.8 %. The nitrogen removal rate and nitrogen removal efficiency at 10.6 °C could maintain at 0.12 kgN/m 3 ·d -1 and 92.5 %, respectively. Microbial analysis reveals that Nitrosomonas, Thauera, and Candidatus&#95;Kuenenia are the predominant genera responsible for nitrite supply and nitrogen removal, localized within the gas channels of granules, flocs, and micro-granules, respectively. Keeping the influent C/NO 3 - -N ratio below 1.7 is ideal to prevent overgrowth of Thauera and maintain system stability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A comprehensive understanding of the defense strategies against nitrite by Lactiplantibacillus plantarum remains unknown. Herein, the effects of nitrite degradation process on metabolic profiling of L. plantarum PK25 were investigated by metabolomics and phenomenological measurement. A total of 633 metabolites were significantly different at 6, 12, and 24 incubation hours. Specifically, citrulline and lysine reduction facilitated strain survival by limiting cell growth. A significant reduction of unsaturated fatty acids was observed, which could induce reduced cell membrane fluidity to prevent nitrite entry. The accumulation of thymine and cytosine might be resulted from accelerated RNA expression to accelerate the repair of cells. Dopamine and ergothioneine could serve as antioxidants to prevent bacteria from oxidative stress. Furthermore, cell filamentation production, increased hydrophobicity, and altered antioxidant enzyme activity were favorable alterations made by strain. Our study demonstrated the metabolite profile alteration of L. plantarum during nitrite degradation, which provided a theoretical basis for targeting strain function., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Challenges in obtaining stable nitrite have impeded the use of anammox in municipal wastewater treatment. This study explored the feasibility of using sludge fermentation products as carbon source and selective nitrification inhibitor to supply nitrite via partial nitrification (PN) and partial denitrification (PD). PD was initiated within 15 days, achieving nitrite transformation rate of over 90 % with a carbon/nitrogen ratio of 3 and a reaction time of 0.75 h. The dominant genus, Romboutsia, increased in relative abundance from 4.1 to 35 %. Organic acids in sludge fermentation products, like acetate (200 mg/L) and propionate (400 mg/L), selectively suppressed nitrite-oxidizing bacteria (NOB) more than ammonia-oxidizing bacteria (AOB), leading to PN. Combining anaerobic exposure with sludge fermentation products addition achieved PN with over 80.0 % nitrite accumulation. AOB increased tenfold in the long term, significantly outpacing NOB growth. This strategy simplifies difficulty of anammox application and shows broad application potential in municipal wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cupric ions can restrain biological nitrogen removal processes, which comprise nitrite reductase and nitric oxide reductase. Here, Pseudomonas sp. NY1 can efficiently perform heterotrophic nitrification and aerobic denitrification with cupric ions at 15 °C. At optimal culturing conditions, low cupric ion levels accelerated nitrogen degradation, and ammonium and nitrite removal efficiencies increased by 2.33%-4.85% and 6.76%-12.30%, respectively. Moreover, the maximum elimination rates for ammonium and nitrite increased from 9.48 to 10.26 mg/L/h and 6.20 to 6.80 mg/L/h upon adding 0.05 mg/L cupric ions. Additionally, low cupric ion concentrations promoted electron transport system activity (ETSA), especially for nitrite reduction. However, high concentrations of cupric ions decreased the ETSA during nitrogen conversion processes. The crucial enzymes ammonia monooxygenase, nitrate reductase, and nitrite reductase possessed similarly trends as ETSA upon exposure to cupric ion. These findings deepen the understanding for the effect of cupric ions on nitrogen consumption and bioremediation in nitrogen-polluted waters., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: This work was financially supported by the National Natural Science Foundation of China (No. 42,167,019 and 42007223). Scientific Research Inovation Team Project of Guizhou University [No. Guidakechuangtuan (2024) 06]., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The partial nitritation-anammox process based on a membrane-aerated biofilm reactor (MABR) faces several challenges, such as difficulty in suppressing nitrite-oxidizing bacteria (NOB), excessive effluent nitrate, and ineffective synergy between denitrification and anammox bacteria. Therefore, a novel partitioned granular sludge coupling with MABR (G-MABR) was constructed. The chemical oxygen demand (COD) and nitrogen removal efficiency were 88.8 ± 1.8 %-92.6 ± 1.2 % and 88.8 ± 1.5 %-93.6 ± 0.7 %, respectively. The COD was mainly lowered in the lower granular sludge-zone, while nitrogen was removed in the upper MABR-zone. NOB was significantly suppressed in the MABR-zone due to competition for substrate with denitrifying bacteria and anammox bacteria. This partitioned configuration reduced the C/N ratio in the MABR-zone, thus facilitating autotrophic nitrogen removal. Both partial nitrification and denitrification provided nitrite for anammox bacteria in granular sludge, whereas partial nitrification mainly supplied nitrite to the anammox bacteria in membrane biofilms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The function of Litopenaeus vannamei Na + /K + /2Cl - cotransporter 1 (NKCC1) under nitrite stress was investigated. The full-length cDNA sequence of the L. vannamei NKCC1 gene was cloned using the rapid amplification of cDNA ends (RACE) technique, and the sequence was analysed using bioinformatics tools. Expression and localisation of NKCC1 in tissues were assessed using real-time quantitative PCR and in situ hybridisation, respectively. The impact of nitrite stress on the survival, physiology, biochemistry and tissue structure of L. vannamei was investigated following silencing of NKCC1 by RNA interference. The 3143 bp cDNA sequence of L. vannamei NKCC1 encodes a polypeptide of 918 amino acids. It is evolutionarily conserved. NKCC1 expression was highest in gill tissue, particularly within cuticle and gill epithelial cells. After silencing NKCC1, an increase in shrimp survival was observed, accompanied by a significant reduction in nitrite entry into the body (P < 0.05). Moreover, the oxidative stress enzyme system remained unaffected and damage to gill tissue was alleviated. The results suggest that NKCC1 is involved in regulating nitrite uptake, and plays a crucial role in facilitating nitrite entry into the organism through gill tissue. The findings provide a vital experimental basis for addressing concerns related to nitrite toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interest or personal relationship that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The explosive and biorefractory nature of nitrocellulose (NC) poses major risks to both humans and the environment. Expanding the range of microorganisms capable of degrading NC is essential, though the most effective known microorganisms, Desulfovibrio genera and Fusarium solani, achieve degradation rates of 5%-25%. Here, a novel strain, Rhodococcus pyridinivorans LZ1 was isolated, demonstrating the ability to degrade NC, with its growth potentially enhanced by the presence of NC. The degradation process was monitored by assessing changes in nitrate, nitrite, and ammonium. Notably, the -OH strength of NC increased over time, whereas the energetic functional groups (-NO 2 and O-NO 2 ) diminished. Furthermore, the presence of NC enhanced nitrate esterase activity 1-2-fold, indicating that ammonification was the primary pathway for NC biodegradation. By converting the nitrate ester of NC into hydroxyl, R. pyridinivorans LZ1 mitigates the harmful effects of NC, offering a promising approach for the treatment of NC waste and wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The aerobic granular sludge (AGS) process had been extensively studied for its simultaneous nitrification and denitrification (SND) capabilities. Iron-carbon (IC) had enhanced AGS nitrogen removal efficiency, but the mechanism remained unclear. In this study, four reactors had been added with 50, 30, 10, and 0 g/L of IC. Total nitrogen removal efficiency increased with IC dosage under the same operation mode. IC enhanced sludge ammonia oxidation rate, denitrification rate, and specific oxygen uptake rate, allowing SND to complete 60 min earlier, potentially reducing wastewater treatment costs. Notably, IC eliminated nitrite accumulation in conventional AGS effluent. IC decreased the abundance of genes and enzyme activities related to NOR expression, while increasing those related to NOS, which may mitigate the potential for nitrous oxide formation by microorganisms. In this study, IC acted as an enzymatic reaction activator, affecting granules more than flocs, with the activity gap gradually decreasing with the IC dosage., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Background: The oral microbiome-dependent nitrate (NO 3 - )-nitrite (NO 2 - )-nitric oxide (NO) pathway may help regulate blood pressure. NO 2 - -producing bacteria in subgingival plaque are reduced in relative abundance in patients with untreated periodontitis compared with periodontally healthy patients. In periodontitis patients, the NO 2 - -producing bacteria increase several months after periodontal treatment. The early effects of periodontal treatment on NO 2 - -producing bacteria and the NO 3 - -NO 2 - -NO pathway remain unknown. The aim of this study was to determine how periodontal treatment affects the oral NO 2 - -producing microbiome and salivary NO 3 - and NO 2 - levels over time., Methods: The subgingival microbiota of 38 periodontitis patients was analysed before (baseline [BL]) and 1, 7 and 90 days after periodontal treatment. Changes in NO 2 - -producing bacteria and periodontitis-associated bacteria were determined by 16s rRNA Illumina sequencing. Saliva samples were collected at all-time points to determine NO 3 - and NO 2 - levels using gas-phase chemiluminescence., Results: A significant increase was observed in the relative abundance of NO 2 - -producing species between BL and all subsequent timepoints (all p < 0.001). Periodontitis-associated species decreased at all timepoints, relative to BL (all p < 0.02). NO 2 - -producing species negatively correlated with periodontitis-associated species at all timepoints, with this relationship strongest 90 days post-treatment (ρ = -0.792, p < 0.001). Despite these findings, no significant changes were found in salivary NO 3 - and NO 2 - over time (all p > 0.05)., Conclusions: Periodontal treatment induced an immediate increase in the relative abundance of health-associated NO 2 - -producing bacteria. This increase persisted throughout periodontal healing. Future studies should test the effect of periodontal treatment combined with NO 3 - intake on periodontal and cardiovascular health., (© 2024 The Author(s). Molecular Oral Microbiology published by John Wiley & Sons Ltd.)

The unique bacterial infection microenvironment (IME) usually requires complicated design of nanomaterials to adapt to IME for enhancing antibacterial therapy. Here, an alternative IME adaptative nitrite reductase-mimicking nanozyme is constructed by in situ growth of ultrasmall copper sulfide clusters on the surface of a nanofibrillar lysozyme assembly (NFLA/CuS NHs), which can temporally regulate nitric oxide (NO) gradient concentration to kill bacteria initially and promote tissue regeneration subsequently. Benefiting from a copper nitrite reductase (CuNIR)-inspired structure with CuS cluster as active center and NFLA as skeleton, NFLA/CuS NHs efficiently boost the catalytic reduction of nitrite to NO. The inherent supramolecular fibrillar networks displays excellent bacterial capture capability, facilitating initial high-concentration NO attacks on the bacteria. The subsequent catalytic release of low-concentration NO by NFLA/CuS NHs-mediated nitrite reduction remarkably promotes cell migration and angiogenesis. This work paves the way for dynamically eliminating MDR bacterial infection and promoting tissue regeneration in a simple and smart way through CuNIR-mimicking catalysis.

Partial nitrification (PN) represents an energy-efficient bioprocess; however, it often confronts challenges such as unstable nitrite accumulation, nitrite oxidizing bacteria shocks, and slow reaction rate. This study established an acidophilic PN with self-sustained pH as low as 5.36. Over 120-day monitoring, nitrite accumulation rate (NAR) was stabilized at more than 97.9 %, and an ultra-high ammonia oxidation rate of 0.81 kg/m 3 ·d was achieved. Notably, least NAR of 77.8 % persisted even under accidental nitrite oxidizing bacteria invasion, aeration delay, alkalinity fluctuations, and cooling shocks. During processing, despite detrimental effects on bacterial diversity, there was a discernible increase in acid-tolerant bacteria responsible for nitrification. Candidatus Nitrosoglobus, gradually dominated in nitrifying guild (2.15 %), with the substantially reduction or disappearance of typical nitrifying microorganisms. Acidobacteriota, a key player in nitrogen cycling of soil, significantly increased from 0.45 % to 9.98 %, and its associated nitrogen metabolism genes showed a substantial 215 % rise. AmoB emerged as the most critical functional gene influencing acidophilic PN, exhibiting significantly higher unit gene expression than other nitrification genes. Blockade tricarboxylic acid cycle, DNA damage, and presence of free nitrous acid exert substantial effects on nitrite-oxidizing bacteria (NOB), serving as internal driving forces for acidophilic PN. This highlights the reliable potential for shortening nitrogen transformation process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) is an important methane (CH 4 ) consumption and nitrogen (N) removal pathway in estuarine and coastal wetlands. Antibiotic contamination is known to affect microbially mediated processes; however, its influences on n-DAMO and the underlying molecular mechanisms remain poorly understood. In the present study, using 13 CH 4 tracer method combined with molecular techniques, we investigated the responses of n-DAMO microbial abundance, activity, and the associated microbial community composition to sulfamethazine (SMT, a sulfonamide antibiotic, with exposure concentrations of 0.05, 0.5, 5, 20, 50, and 100 µg L -1 ). Results showed that the effect of SMT exposure on n-DAMO activity was dose-dependent. Exposure to SMT at concentrations of up to 5 µg L -1 inhibited the potential n-DAMO rates (the average rates of nitrite- and nitrate-DAMO decreased by 92.9 % and 79.2 % relative to the control, respectively). In contrast, n-DAMO rates tended to be promoted by SMT when its concentration increased to 20-100 µg L -1 (the average rates of nitrite- and nitrate-DAMO increased by 724.1 % and 630.1 % relative to the low-doses, respectively). Notably, low-doses of SMT suppressed nitrite-DAMO to a greater extent than nitrate-DAMO, indicating that nitrite-DAMO was more sensitive to SMT than nitrate-DAMO. Molecular analyses suggest that the increased n-DAMO activity under high-doses SMT exposure may be driven by changes in microbial communities, especially because of the promotion of methanogens that provide more CH 4 to n-DAMO microbes. Moreover, the abundances of n-DAMO microbes at high SMT exposure (20 and 50 µg L -1 ) were significantly higher than that at low SMT exposure (0.05-5 µg L -1 ). These results advance our understanding of the ecological effects of SMT on carbon (C) and N interactions in estuarine and coastal wetlands., Competing Interests: Declaration of Competing Interest The authors declare no known conflicts of competing financial interest or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)