Deciphering behaviors of 6:2 chlorinated polyfluorinated ether sulfonate (alternative-PFOS) on anammox processes: Nitrogen removal efficiency and microbial adaptability.

This study investigates the behaviors and effects of F-53B, an alternative to perfluorooctane sulfonate on anaerobic ammonium oxidation (anammox) processes. Results showed that the nitrogen removal efficiency (NRE) reached 83.8 % at a F-53B concentration of 0.5 mg·L-1, while NRE decreased to 66.9 % with 5 mg·L-1 of F-53B. The defluorination rates of 17.8 % (0.5 mg·L-1) and 9.3 % (5 mg·L-1) were observed, respectively, suggesting the occurrence of F-53B degradation. The relative abundance of Ca. Kuenenia decreased from 26.1 % to 16.2 % with the F-53B concentration increasing from 0.5 mg·L-1 to 5 mg·L-1. Meanwhile, Denitratisoma was selectively enriched with a relative abundance of 40.7 % at an F-53B concentration of 0.5 mg·L-1. Ca. Kuenenia could reduce reactive oxygen species induced by F-53B to maintain the balance of oxidative stress. This study gains insight into the behaviors and metabolic mechanisms of F-53B in anammox consortia, suggesting the feasibility of anammox processes for industrial wastewater.

Multi-omics analysis reveals the collaboration and metabolisms of the anammox consortia driven by soluble/non-soluble Fe(III) as the sole iron element.

Iron is recognized as a physiological requirement for anammox bacteria (AnAOB), with Fe(II) considered to be the most effective form. However, Fe(III), instead of Fe(II) is the common iron form in natural and artificial ecosystems. In this study, the nitrogen removal performance and metabolic mechanisms in anammox consortia with soluble and non-soluble Fe(III) as the sole iron element were investigated. After the 150-day operation, the soluble (FeCl3) and insoluble (Fe2O3) Fe(III)-fed anammox systems reached nitrogen removal rates of 71.84 ± 0.80% and 50.20 ± 0.98%, respectively. AnAOB could survive with soluble (FeCl3) or insoluble (Fe2O3) Fe(III) as the sole iron element, reaching relative abundances of 18.49% and 13.16%, respectively. The results show that the formation of anammox core consortia can enable AnAOB's survival to adverse external conditions of Fe(II) deficiency. Metagenomic and metatranscriptomic analysis reveal that Ca. Kuenenia can only uptake Fe(II) into the cell for metabolisms either independently through the extracellular electron transfer or with the cross-feeding of symbiotic microbes. This study provides insight into the utilization and metabolic mechanisms of Fe(III) in Ca. Kuenenia-dominated consortia, and deepens the understanding of anammox core consortia in the nitrogen, carbon, and iron cycling, further promoting the practical applications of anammox processes.

Light-Driven Electron Uptake from Nonfermentative Organic Matter to Expedite Nitrogen Dissimilation by Chemolithotrophic Anammox Consortia.

Nonphotosynthetic microorganisms are typically unable to directly utilize light energy, but light might change the metabolic pathway of these bacteria indirectly by forming intermediates such as reactive oxygen species (ROS). This work investigated the role of light on nitrogen conversion by anaerobic ammonium oxidation (anammox) consortia. The results showed that high intensity light (>20000 lx) caused ca. 50% inhibition of anammox activity, and total ROS reached 167% at 60,000 lx. Surprisingly, 200 lx light was found to induce unexpected promotion of the nitrogen conversion rate, and ultraviolet light (<420 nm) was identified as the main contributor. Metagenomic and metatranscriptomic analyses revealed that the gene encoding cytochrome c peroxidase was highly expressed only under 200 lx light. 15N isotope tracing, gene abundance quantification, and external H2O2 addition experiments showed that photoinduced trace H2O2 triggered cytochrome c peroxidase expression to take up electrons from extracellular nonfermentative organics to synthesize NADH and ATP, thereby expediting nitrogen dissimulation of anammox consortia. External supplying reduced humic acid into a low-intensity light exposure system would result in a maximal 1.7-fold increase in the nitrogen conversion rate. These interesting findings may provide insight into the niche differentiation and widespread nature of anammox bacteria in natural ecotopes.

Microbial interactions play a keystone role in rapid anaerobic ammonium oxidation sludge proliferation and biofilm formation.

Two mature anaerobic ammonium oxidation (anammox) consortia with high/low relative abundance of anammox bacteria were inoculated for the rapid sludge proliferation and biofilm formation in this study, named up-flow anaerobic sludge blanket reactor (UASB1) (high) and UASB2 (low), respectively. Results showed that the nitrogen removal efficiency of UASB2 reached 90.94% after the 120-day operation, which was 13% higher than that of UASB1. Moreover, its biomass amounts were 22.18% (biofilm) and 40.96% (flocs) higher than that of UASB1, respectively. Ca. Kuenenia possessed relative abundances of 29.32% (flocs), 27.42% (biofilm) and 31.56% (flocs), 35.20% (biofilm) in the UASB1 and UASB2, respectively. The relative abundances of genes involved in anammox transformation (hzs, nir) and carbon metabolism (fdh, lgA/B/C, acs) were higher in the UASB2, indicating that Ca. Kuenenia might produce acetate and glycogen to enhance microbial interactions. These findings emphasized the importance of microbial interactions in anammox sludge proliferation and biofilm formation.

Biosynthesis of poly(3-hydroxybutyrate) by N,N-dimethylformamide degrading strain Paracoccus sp. PXZ: A strategy for resource utilization of pollutants.

N,N-dimethylformamide is a toxic chemical solvent, which widely exists in industrial wastewater. Nevertheless, the relevant methods merely achieved non-hazardous treatment of N,N-dimethylformamide. In this study, one efficient N,N-dimethylformamide degrading strain was isolated and developed for pollutant removal coupling with poly(3-hydroxybutyrate) (PHB) accumulation. The functional host was characterized as Paracoccus sp. PXZ, which could consume N,N-dimethylformamide as the nutrient substrate for cell reproduction. Whole-genome sequencing analysis confirmed that PXZ simultaneously possesses the essential genes for poly(3-hydroxybutyrate) synthesis. Subsequently, the approaches of nutrient supplementation and various physicochemical variables to strengthen poly(3-hydroxybutyrate) production were investigated. The optimal biopolymer concentration was 2.74 g·L-1 with a poly(3-hydroxybutyrate) proportion of 61%, showing a yield of 0.29 g-PHB·g-1-fructose. Furthermore, N,N-dimethylformamide served as the special nitrogen matter that could realize a similar poly(3-hydroxybutyrate) accumulation. This study provided a fermentation technology coupling with N,N-dimethylformamide degradation, offering a new strategy for resource utilization of specific pollutants and wastewater treatment.

Succession of microbial communities reveals the inevitability of anammox core in the development of anammox processes.

The lack of anammox seeds is regarded as the bottleneck of anammox-based processes. Although the interactions in anammox consortia have attracted increasing attention, little is known about the influence of inoculated sludge populations on the growth of anammox bacteria. In this study, four sludge of distinct communities mixed with anammox sludge (the relative abundance of Ca. Kuenenia was 1.96 %) were used as the seeds, respectively for the start-up of anammox processes. Notably, all these mixed microbial communities tend to form a similar microbial community, defined as the anammox core, containing anammox-bacteria (22.9 ± 5.9 %), ammonia-oxidizing-bacteria (0.8 ± 0.7 %), nitrite-oxidizing-bacteria (0.2 ± 0.2 %), Chloroflexi-bacteria (0.7 ± 0.4 %), and heterotrophic-denitrification-bacteria (0.3 ± 0.2 %). It also elucidated that the communities of Nitrosomonas-dominated sludge were the closest to the anammox core, and achieved the highest nitrogen-removal rate of 0.73 kg-N m-3 d-1. This study sheds light on the solution to the shortage of anammox seeds in the full-scale wastewater treatment application.

Coupling continuous poly(3-hydroxybutyrate) synthesis with piperazine-contained wastewater treatment: Fermentation performance and microbial contamination deciphering.

Open poly(3-hydroxybutyrate) (PHB) fermentation is of great potential, and batch PHB synthesis with piperazine as the nitrogen switch has been realized. However, it is vital to explore the feasibility of continuous PHB fermentation with piperazine-contained wastewater remediation collaboratively. Here, an aerobic membrane bioreactor was constructed for consecutive PHB synthesis. The removal efficiency of piperazine decreased from 100 % to 82.6 % after three cycles, meanwhile, the PHB concentration was 0.39 g·L-1, 0.18 g·L-1, and undetected for each cycle. Microbial community analysis showed that Proteobacteria, Actinobacteriota, and Bacteroidota were the main contaminating microbes. Furthermore, three metagenome-assembled genomes related to Flavobacterium collumnare, Herbaspirillum aquaticum, and Microbacterium enclense were identified as the dominant contaminating strains. These microbes obtained nitrogenous substrates transformed by Paracoccus sp. TOH, such as amino acids and dissolved organic matter, as nutrient for accumulation. This study verified the practicability of coupling continuous PHB synthesis with industrial wastewater treatment and revealed the derivation mechanism of contaminating species, which could provide a reference for the targeted nitrogen release gene knockout of functional PHB fermentation chassis.

The organics-mediated microbial dynamics and mixotrophic metabolisms in anammox consortia under micro-aerobic conditions.

The engineering applications of mainstream anaerobic ammonium oxidation (anammox) have raised increasing attention due to its energy-efficient, however, the organics-mediated microbial dynamics and mixotrophic metabolisms in anammox consortia under micro-aerobic conditions are still elusive. Here, the response of the anammox process to sodium acetate and glucose at a C/N ratio ranging from 0 to 0.5 was investigated under micro-aerobic conditions, respectively. Results showed that the additional glucose could promote the nitrogen removal efficiency (NRE) and nitrogen removal rate (NRR) of anammox processes at a low C/N ratio (0.3), representing 84.00% and 0.53 N kg·m-3·d-1. The introduced organics could regulate the diversity of the microbial community and simplify the microbial relationship in anammox consortia. Anammox could not benefit from the introduced sodium acetate, while glucose could effectively enhance the anammox activity and microbial interactions in anammox consortia. Glucose might also stimulate the mixotrophic mechanism of Ca. Kuenenia, further promotes the proliferation of anammox sludge under micro-aerobic conditions. This study reveals that glucose could positively mediate microbial interactions and mixotrophic metabolism in anammox consortia under micro-aerobic conditions, which raises a new horizon for the proliferation of anammox sludge for mainstream engineering applications.

Bioconversion of waste activated sludge hydrolysate into polyhydroxyalkanoates using Paracoccus sp. TOH: Volatile fatty acids generation and fermentation strategy.

The expensive carbon matrix is a bottleneck restricting the industrialization of polyhydroxyalkanoates (PHAs). Volatile fatty acids (VFAs) derived from waste activated sludge via anaerobic fermentation might be alternative carbon matters for PHAs synthesis. In this study, the effect of enzymes on VFAs yields and the feasibility of the produced VFAs for PHAs fermentation by Paracoccus sp. TOH were investigated. The optimum cumulative VFAs concentration reached 4076.6 mg-COD·L-1 in the lysozyme treatment system. Correspondingly, the highest poly(3-hydroxybuturate-co-3-hydroxyvalerate) (PHBV) concentration (119.1 mg·L-1) containing 20.3 mol% 3-hydroxyvalerate was obtained. It proved that Paracoccus sp. TOH possesses the capability for PHBV accumulation. The functional hydrolytic-acidogenic microorganisms, such as Clostridium sensu stricto and Bacteroides sp. were accumulated. The functional genes encoding hydrolysis, carbohydrates metabolism, VFAs generation were enriched. This study offered a possible strategy for VFAs production and verified the feasibility of sludge hydrolysate as a high-quality carbon substrate for PHAs fermentation.

Microbial dynamics reveal the adaptation strategies of ecological niche in distinct anammox consortia under mainstream conditions.

The feasibility of anammox-based processes for nitrogen-contained wastewater treatment has been verified with different anammox bacteria, however, the ecological niche of anammox bacteria under mainstream conditions is still elusive. In this study, six sludge samples collected from different habitats were utilized to culture anammox bacteria under mainstream conditions, and two distinct anammox genera (Ca. Kuenenia and Ca. Brocadia) with a relative abundance of 6.31% (C1) and 3.09% (C3), respectively, were identified. Notably, the microbial dynamics revealed that anammox bacteria (AMX), ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), Chloroflexi bacteria (CFX), and heterotrophic denitrification bacteria (HDB) were the core members in anammox consortia. However, Ca. Kuenenia and Ca. Brocadia occupied different ecological niches in anammox consortia. The dissolved oxygen and microbial structures of the anammox-continuous stirred tank reactor systems were the main factors to affect their niche differentiation. Meanwhile, comammox might exist in the systems and occupy the ecological niche of AOB in nitrogen cycling. The network analysis suggested that Ignavibacterium could be the associated bacteria in Ca. Kuenenia-dominated consortia, while Ca. Nitrotoga was that in the Ca. Brocadia-dominated consortia. Our findings reveal a valuable reference for the observation of distinct anammox genera under mainstream conditions, which provides theoretical guidance for the engineering application of mainstream anammox-based processes.

Nitrogen contribution and microbial community of size-fractionated anammox sludge in continuous stirred-tank reactors.

In this study, the microbial diversity of size-fractionated anammox sludge in a well-mixed system and their contribution to nitrogen transformation were investigated. Results showed that small granules (0.2-1.0 mm) contributed to the major part of the nitrogen removal rate (56 %) due to its largest mixed liquor volatile suspended solids (1240 ± 80 mg·L-1). However, large granules (>1.0 mm) possessed the highest relative abundances of Ca. Kuenenia stuttgartiensis and specific anammox activity, representing 49.34 % and 24.45 ± 0.01 mg-N·g-1-mixed liquor volatile suspended solids·h-1, respectively. The microbial diversity decreased as the increase of granular size, resulting in microbial community shifting to a simpler model. Metagenomic analysis showed that fine sludge might be the potential major for NO/N2O production in the mature well-mixed system under inorganic conditions. This study provides guidance for the evaluation of nitrogen contribution by anammox size-fractionated sludge and the inhibition of the potential NO/N2O emission in anammox processes.

Synthesis of tetrasubstituted allenes via a 1,4-palladium migration/carbene insertion/ß-H elimination sequence.

A palladium-catalyzed synthesis of tetrasubstituted allenes from aryl bromides and aryl diazoacetates is developed. This transformation proceeded via an aryl to alkenyl 1,4-palladium migration/carbene insertion/ß-H elimination sequence under mild reaction conditions.

Palladium-catalyzed cross-coupling of unreactive C(sp3)-H bonds with azole C(sp2)-H bonds by using bromide as a traceless directing group.

A palladium-catalyzed intermolecular cross-coupling of unreactive C(sp3)-H bonds and azole C(sp2)-H bonds with bromide as a traceless directing group is described. The judicious selection of the bulky and electron-rich phosphine ligand is the key for the success of this cascade process. The protocol features a broad substrate scope, excellent regioselectivity, and good functional group tolerance.

Phosphorylation of C(sp3)-H Bonds via 1,4-Palladium Migration.

1,4-Palladium migration has emerged as a reliable method for directed C-H functionalization. In contrast to coupling with carbon nucleophiles, limited examples with heteroatom nucleophiles have been reported. Herein we report a palladium-catalyzed intermolecular C(sp3)-H phosphorylation reaction via 1,4-palladium migration, which is often difficult because of the strong coordination of phosphorus reagents to palladium catalysts. Phosphorylation of C(sp3)-H bonds is accomplished in good reaction yields with excellent regioselectivity. The judicious selection of the phosphine ligand proved to be the key to the success of this cascade process.

How anammox responds to the emerging contaminants: Status and mechanisms.

Numerous researches have been carried out to study the effects of emerging contaminants in wastewater, such as antibiotics, nanomaterials, heavy metals, and microplastics, on the anammox process. However, they are fragmented and difficult to provide a comprehensive understanding of their effects on reactor performance and the metabolic mechanisms in anammox bacteria. Therefore, this paper overviews the effects on anammox processes by the introduced emerging contaminants in the past years to fulfill such knowledge gaps that affect our perception of the inhibitory mechanisms and limit the optimization of the anammox process. In detail, their effects on anammox processes from the aspects of reactor performance, microbial community, antibiotic resistance genes (ARGs), and functional genes related to anammox and nitrogen transformation in anammox consortia are summarized. Furthermore, the metabolic mechanisms causing the cell death of anammox bacteria, such as induction of reactive oxygen species, limitation of substrates diffusion, and membrane binding are proposed. By offering this review, the remaining research gaps are identified, and the potential metabolic mechanisms in anammox consortia are highlighted.

[Effect of Biochar on Soil Enzyme Activity & the Bacterial Community and Its Mechanism].

Biochar-based fertilizers can improve the mineralization of carbon and nitrogen in soil and enhance the soil micro-ecological environment due to particular physical and chemical properties. It is of great significance to explore the underlying mechanism of biochar-based fertilizer in the regulation of soil microorganisms and soil enzyme activity to improve soil quality. Field experiments were conducted to investigate the effects of different biochar-based fertilizer rates[0 (CK2), 0.6 (T1), 0.9 (T2), 1.2 (T3), and 1.5 (T4) t·hm-2]on soil nutrients, soil enzyme activity, and bacterial community structure. The results showed that with the application of biochar-based fertilizer, soil bulk density decreased, while the pH value, available P, available K, organic matter content, and the C/N ratio increased by 0.32%-5.83%, 14.09%-23.16%, 0%-38.70%, 7.49%-14.16%, and 4.06%-10.13%, respectively, compared to that of the CK2 treatment. With increasing rates of biochar-based fertilizer, the enzyme activity first increased and then decreased. Invertase (INV), urease (URE), catalase (CAT), and neutral phosphatase (NPH) activity under the application of biochar-based fertilizer were 63.73%-166.37%, 117.52%-174.03%, 12.98%-23.59%, and 60.84%-119.71% higher than that of CK2, respectively. The corresponding bacterial diversity was significantly improved, especially with regard to the increase in the abundance of growth promoting bacteria, such as Gemmatimonadetes and Proteobacteria, and decreased the abundance of Acidobacteria and Actinobacteria. The correlation analysis showed that soil C/N ratio was the key factor affecting soil enzyme activity, and there was a significant positive correlation between soil enzyme activity and bacterial diversity. There were significantly positive correlations among the activities of the above four soil enzymes and the relative abundance of Gemmatimonadetes (P<0.01), with CAT being the key factor affecting the bacterial community structure. This study revealed a relationship between soil enzyme activity and microbial colonies, which provides a theoretical basis and mechanism for applying biochar to regulate the soil enzyme and micro-ecological environment.

Evolution of microbial community and antibiotic resistance genes in anammox process stressed by oxytetracycline and copper.

The individual and combined impacts of copper ion (Cu2+) and oxytetracycline (OTC) on anaerobic ammonium oxidation (anammox) performance and its self-recovery process were examined. Experimental results showed that the anammox performance and activity of anammox bacteria were inhibited by 1.0 mg L-1 OTC, Cu2+ and OTC + Cu2+, and both single and combined inhibitions were reversible. The abundance of functional genes and parts of antibiotic resistance genes (ARGs) were positively related to the dominant bacterium Ca. Kuenenia, implying that the recovery of the performance was associated with the progressive induction of potentially resistant species after inhibition. The above outcomes illustrated that anammox bacteria were stressed by metals and antibiotics, but they still could remove nitrogen at a rate higher than 20.6 ± 0.8 kg N m-3 d-1, providing guidance for engineering applications of anammox processes.

Perfluorooctane sulfonate disrupts the blood brain barrier through the crosstalk between endothelial cells and astrocytes in mice.

Perfluorooctane sulfonate (PFOS), a classic environmental pollutant, is reported to accumulate in brain and induce neurotoxicity. However, little is known the route and mechanism of its entrance in brain. In the present study, ICR mice were treated with PFOS for 28 days, the cerebral PFOS were measured and the morphological and ultrastructural changes of blood-brain barrier (BBB) were observed. Also, the expression and localization of the proteins related to the cerebral damages, tight junctions (TJs) and p38 activation were detected. Additionally, U87 cells were used to explore the role of p38 in PFOS-induced damages of astrocytes. PFOS significantly decreased the expression of TJ-related proteins (ZO-1, Claudin-5, Claudin-11, Occludin) in endothelial cells and disrupted BBB, which subsequently led PFOS to astrocytes and increased the expression of the proteins related to astrocytic damages (Aquaporin 4 and S100ß). These results aggravated BBB disruption and further increased the cerebral PFOS levels. Besides, phosphorylated p38 activation was involved into PFOS-induced astrocytic damages in vivo and in vitro. In conclusion, the crosstalk between endothelial cells and astrocytes facilitated the BBB disruption and increased the accumulation of PFOS in brain. Our findings provided a new insight into the toxicological and physiological profiles of PFOS-induced neurotoxicity.

Finasteride inhibited brain dopaminergic system and open-field behaviors in adolescent male rats.

AIMS: Finasteride inhibits the conversion of testosterone to dihydrotestosterone. Because androgen regulates dopaminergic system in the brain, it could be hypothesized that finasteride may inhibit dopaminergic system. The present study therefore investigates the effects of finasteride in adolescent and early developmental rats on dopaminergic system, including contents of dopamine and its metabolites (dihydroxy phenyl acetic acid and homovanillic acid) and tyrosine hydroxylase expressions both at gene and protein levels. Meanwhile, open-field behaviors of the rats are examined because of the regulatory effect of dopaminergic system on the behaviors. METHODS: Open-field behaviors were evaluated by exploratory and motor behaviors. Dopamine and its metabolites were assayed by liquid chromatography-mass spectrometry. Tyrosine hydroxylase mRNA and protein expressions were determined by real-time qRT-PCR and western blot, respectively. RESULTS: It was found that in adolescent male rats, administration of finasteride at doses of 25 and 50 mg/kg for 14 days dose dependently inhibited open-field behaviors, reduced contents of dopamine and its metabolites in frontal cortex, hippocampus, caudate putamen, nucleus accumbens, and down-regulated tyrosine hydroxylase mRNA and protein expressions in substantia nigra and ventral tegmental area. However, there was no significant change of these parameters in early developmental rats after finasteride treatment. CONCLUSION: These results suggest that finasteride inhibits dopaminergic system and open-field behaviors in adolescent male rats by inhibiting the conversion of testosterone to dihydrotestosterone, and imply finasteride as a potential therapeutic option for neuropsychiatric disorders associated with hyperactivities of dopaminergic system and androgen.

[Application progress of proteomic in pharmacological study of Chinese medicinal formulae].

Chinese medicinal formulae are the important means of clinical treatment in traditional Chinese medicine. It is urgent to use modern advanced scientific and technological means to reveal the complicated mechanism of Chinese medicinal formulae because they have the function characteristics of multiple components, multiple targets and integrated regulation. The systematic and comprehensive research model of proteomic is in line with the function characteristics of Chinese medicinal formulae, and proteomic has been widely used in the study of pharmacological mechanism of Chinese medicinal formulae. The recent applications of proteomic in pharmacological study of Chinese medicinal formulae in anti-cardiovascular and cerebrovascular diseases, anti-liver disease, antidiabetic, anticancer, anti-rheumatoid arthritis and other diseases were reviewed in this paper, and then the future development direction of proteomic in pharmacological study of Chinese medicinal formulae was put forward. This review is to provide the ideas and method for proteomic research on function mechanism of Chinese medicinal formulae.