Architecture of chloroplast TOC-TIC translocon supercomplex.

Chloroplasts rely on the translocon complexes in the outer and inner envelope membranes (the TOC and TIC complexes, respectively) to import thousands of different nuclear-encoded proteins from the cytosol1-4. Although previous studies indicated that the TOC and TIC complexes may assemble into larger supercomplexes5-7, the overall architectures of the TOC-TIC supercomplexes and the mechanism of preprotein translocation are unclear. Here we report the cryo-electron microscopy structure of the TOC-TIC supercomplex from Chlamydomonas reinhardtii. The major subunits of the TOC complex (Toc75, Toc90 and Toc34) and TIC complex (Tic214, Tic20, Tic100 and Tic56), three chloroplast translocon-associated proteins (Ctap3, Ctap4 and Ctap5) and three newly identified small inner-membrane proteins (Simp1-3) have been located in the supercomplex. As the largest protein, Tic214 traverses the inner membrane, the intermembrane space and the outer membrane, connecting the TOC complex with the TIC proteins. An inositol hexaphosphate molecule is located at the Tic214-Toc90 interface and stabilizes their assembly. Four lipid molecules are located within or above an inner-membrane funnel formed by Tic214, Tic20, Simp1 and Ctap5. Multiple potential pathways found in the TOC-TIC supercomplex may support translocation of different substrate preproteins into chloroplasts.

Natural bioink of interpenetrating network hydrogels mimicking extracellular polymeric substances for microbial immobilization in water pollution control.

Artificial biomanufacturing has been developed as a promising biotechnology for water pollution control. Effective bioimmobilization techniques are limited in application because of low productivity and the difficulty in achieving both mechanical strength and biocompatibility. Bioprinting technology, using biomaterials as bioink to enable the rapid on-demand production of bioactive structures, opens a new path for bioimmobilization. In this study, mimicking extracellular polysaccharide and protein of aerobic granular sludge (AGS), sodium alginate (SA) and silk fibroin methacryloyl (SilMA) were developed as the dual-component bioink with a suitable viscosity for bioprinting hydrogel. Interpenetrating network (IPN) hydrogel beads were manufactured using 1.5% (w/v) SA combined with 20% (w/v) SilMA through physical and covalent crosslinking, which exhibited excellent structural stability and bioactivity. The addition of SilMA provided a solution to the poor mechanical stability of SA-Ca hydrogels limited by Ca2+-Na+ ionic exchange. The unique structure of SilMA contributed to the reduction of hydrogel swelling as well as the prevention of SA loss. IPN hydrogels showed a swelling rate of less than 20% compared to the high swelling rate of more than 60% for SA hydrogels. On the other hand, SA controlled the hardening induced by excessive self-assembly of SilMA and improved mass transport in SilMA hydrogels. Compared to IPN hydrogels, SilMA hydrogels experienced a 15% volumetric shrinkage and exhibited a low water content of 92%. Sonication pretreatment of the dual-component bioink not only increased the intermolecular chain entanglement to form IPN, but also led to ß-sheet content in SiMA reaching 46%-48%, which resulted in the formation of stable IPN hydrogels dominated entirely by physical crosslinking. Satisfactory proliferation and viability were achieved for the encapsulated bacteria in IPN hydrogels (µmax 1.49-2.18 d-1). Further, the IPN biohydrogels could maintain structural stability as well as achieve pollutant removal for treating synthetic wastewater with high Na+ concentration of 300 mg/L. The novel SA/SilMA hydrogel bioprinting strategy established in this study offers a new direction for bioimmobilization in water pollution control and other environmental applications.

Ammonia monooxygenase-mediated transformation of 17α-ethinylestradiol: Underlying molecular mechanism.

17α-ethinylestradiol (EE2) has received increasing attention as an emerging and difficult-to-remove emerging contaminant in recent years. Ammonia-oxidizing bacteria (AOB) have been reported to be effective in EE2 removal, and ammonia monooxygenase (AMO) is considered as the primary enzyme for EE2 removal. However, the molecular mechanism underlying the transformation of EE2 by AOB and AMO is still unclear. This study investigated the molecular mechanism of EE2 degradation using a combination of experimental and computational simulation methods. The results revealed that ammonia nitrogen was essential for the co-metabolism of EE2 by AOB, and that NH3 bound with CuC (one active site of AMO) to induce a conformational change in AMO, allowing EE2 to bind with the other active site (CuB), and then EE2 underwent biological transformation. These results provide a theoretical basis and a novel research perspective on the removal of ammonia nitrogen and emerging contaminants (e.g., EE2) in wastewater treatment.

Effects of zero-valent iron (ZVI) on nitrogen conversion, transformation of sulfamethoxazole (SMX) and abundance of antibiotic resistance genes (ARGs) in aerobic granular sludge process.

Even after pre-treatment, livestock and poultry wastewater still contain high concentrations of ammonia and residual antibiotics. These could be removed economically using the aerobic granular sludge (AGS) process with zero-valent iron (ZVI). The interaction of antibiotics and nitrogen in this process needs to be clarified and controlled, however, to achieve good removal performance. Otherwise, antibiotics might generate transformation products (TPs) with higher toxicity and lead to the emergence of antibiotic-resistant bacteria carrying antibiotic resistance genes (ARGs), which could cause persistent toxicity and the risk of disease transmission to the ecological environment. This study investigated the impact of ZVI on AGS for nitrogen and sulfamethoxazole (SMX) removal. The results show that AGS could maintain good ammonia removal performance and that the existence of SMX had a negative impact on ammonia oxidation activities. ZVI contributed to an increase in the abundance of nitrite oxidation bacteria, denitrifying bacteria and the functional genes of nitrogen removal. This led to better total nitrogen removal and a decrease in N2O emission. Accompanied by biological nitrogen transformation, SMX could be transformed into 14 TPs through five pathways. ZVI has the potential to enhance transformation pathways with TPs of lower ecotoxicity, thereby reducing the acute and chronic toxicity of the effluent. Unfortunately, ZVI might enhance the abundance of sul1, sul2, and sul3 in AGS, which increases the risk of sulfonamide antibiotic resistance. In AGS, Opitutaceae, Xanthomonas, Spartobacteria and Mesorhizobium were potential hosts for ARGs. This study provides theoretical references for the interaction of typical antibiotics and nitrogen in the biological treatment process of wastewater and bioremediation of natural water bodies.

Necessity of stirring for outdoor microalgal-bacterial granular sludge process.

As a green process, microalgal-bacterial granular sludge (MBGS) process shows talents in achieving pollutant removal, resource recovery and carbon neutralization. However, when it comes to application, the adequate mixing of MBGS and substrate should be adopted theoretically. Therefore, this study devoted to address the necessity of stirring for MBGS in municipal wastewater treatment. Outdoor performances showed that stirring significantly enhanced both of the photosynthetic efficiency and biomass productivity of MBGS with almost 2-fold increase as compared to non-stirred MBGS, while the average pore size and microalgae-to-bacteria ratio also increased. Consequently, stirring acted as a pivotal role in accelerating pollutants removal, with removals of organics (89.89% COD) and nutrients (99.22% NH4+-N, 92.15% PO43--P) reaching peak levels at 2 h and 6 h, respectively, while removals of organics (87.50% COD) and nutrients (86.11% NH4+-N, 86.76% PO43--P) removal peaked at 8 h for non-stirred MBGS. The improved granule characteristics and microbial compositions due to the stirring were found to be favorable for MBGS to adapting to the changeable weather. Based on the above results, the possible underlying mechanisms of stirring for improving MBGS were illustrated. Overall, stirring positively impacted the photosynthetic efficiency, biomass productivity, pollutant removal and microbial structure for MBGS. This study gains knowledge on stirred MBGS process under outdoor conditions for its future practical application.

A novel electrochemical sensor based on autotropic and heterotrophic nitrifying biofilm for trichloroacetic acid toxicity monitoring.

Trichloroacetic acid (TCA), a toxic substance produced in the disinfection process of wastewater treatment plants, will accumulate in the receiving water. The detection of TCA in the water can achieve the purpose of early warning. However, currently there are few reports on microbial sensors used for TCA detection, and the characteristics of their microbial communities are still unclear. In this work, a toxicity monitoring microbial system (TMMS) with nitrifying biofilm as a sensing element and cathode oxygen reduction as a current signal was successfully constructed for TCA detection. The current and nitrification rate showed a linear relationship with low TCA concentration from 0 to 50 µg/L (R2current = 0.9892, R2nitrification = 0.9860), and high concentration range from 50 to 5000 µg/L (R2current = 0.9883, R2nitrification = 0.9721). High-throughput sequencing revealed that the TMMS was composed of autotrophic/heterotrophic nitrifying and denitrifying microorganisms. Further analysis via symbiotic relationship network demonstrated that Arenimonas and Hyphomicrobium were the core nodes for maintaining interaction between autotropic and heterotrophic nitrifying bacteria. Kyoto Encyclopedia of Genes and Genomes analysis showed that after adding TCA to TMMS, the carbon metabolism and the abundance of the tricarboxylic acid cycle pathway were reduced, and the activity of microorganisms was inhibited. TCA stress caused a low abundance of nitrifying and denitrifying functional enzymes, resulting in low oxygen consumption in the nitrification process, but more oxygen supply for cathode oxygen reduction. This work explored a novel sensor combined with electrochemistry and autotrophic/heterotrophic nitrification, which provided a new insight into the development of microbial monitoring of toxic substances.

Effect of ozone pretreatment on biogranulation with partial nitritation - Anammox two stages for nitrogen removal from mature landfill leachate.

Due to the extremely low C/N ratio, high concentration of ammonia nitrogen and refractory organic matter of mature landfill leachate (MLL), appropriate processes should be selected to effectively remove nitrogen and reduce disposal costs. Partial nitritation (PN) and anaerobic ammonia oxidation (AMX) have been used as the main nitrogen removal processes for MLL, and the sludge granulation in PN and AMX processes could contribute to high biological activity, good sedimentation performance, and stable resistance to toxicity. In this study, the O3-PN-AMX biogranules process was selected to effectively remove nitrogen from MLL without carbon addition and pH adjustment. Without uneconomical NH4+-N oxidation and wasting the alkalinity of MLL, ozone pretreatment achieved color removal, decreased humic- and fulvic-like acid substances, and alleviated the MLL toxicity on ammonia oxidizers. In addition, the ozonation of MLL could shorten the start-up time and improve the treatment efficiency and biogranules stability of PN and AMX processes. Efficient and stable nitritation was achieved in PN reactor without strict dissolved oxygen (DO) control, which was attributed to the unique structure of granular sludge, ozone pretreatment, and alternating inhibition of free ammonia and free nitric acid on nitrite oxidizers. Through the application of ozone pretreatment and granular sludge, the nitrogen removal rate (NRR) and nitrogen removal efficiency (NRE) of the O3-PN-AMX biogranules process reached 0.39 kg/m3/day and 85%, respectively, for the undiluted MLL treatment. This study might provide a novel and effective operation strategy of combined process for the efficient, economical, and stable nitrogen removal from MLL.

Bowel endometriosis: diagnosis and management.

The most common location of extragenital endometriosis is the bowel. Medical treatment may not provide long-term improvement in patients who are symptomatic, and consequently most of these patients may require surgical intervention. Over the past century, surgeons have continued to debate the optimal surgical approach to treating bowel endometriosis, weighing the risks against the benefits. In this expert review we will describe how the recommended surgical approach depends largely on the location of disease, in addition to size and depth of the lesion. For lesions approximately 5-8 cm from the anal verge, we encourage conservative surgical management over resection to decrease the risk of short- and long-term complications.

Synergy of N-(3-oxohexanoyl)-L-homoserine lactone and tryptophan-like outer extracellular substances in granular sludge dominated by aerobic ammonia-oxidizing bacteria.

Nitrogen removal via nitrite is an energy-saving method for high-strength ammonia wastewater treatment. A better understanding of the formation of granular sludge dominated by aerobic ammonia-oxidizing bacteria (AerAOB) could facilitate the improved use of rapid sludge granulation for nitritation. In this study, AerAOB-dominated activated sludge (NAS) and granular sludge (NGS) produced different N-scyl-homoserine lactones (AHLs). N-(3-oxohexanoyl)-L-homoserinelactone (OHHL), only released from NGS, was shown to accelerate sludge aggregation by increasing the biomass growth rate, microbial activity, extracellular protein, and AerAOB biomass. For both NAS and NGS, sludge cells were glued together by inner extracellular polymeric substances (EPSs) with similar components to form microcolony. Different from the characterized negative effect of NAS's outer-EPS on cell adhesion, the outer-EPS of NGS played a positive role in the attached growth of AerAOB-dominated sludge and contained more tryptophan-like substances. More interesting, OHHL enhanced the yields of tryptophan-like substances after mixing with the outer-EPS of NGS, enhancing cell adhesion. In a word, OHHL and more tryptophan-like substances were produced in the process of granulation under the selective sludge discharge condition, which was proved to be able to accelerate NAS granulation. Therefore, the sludge granulation process for nitritation can be improved by increasing the levels of OHHL and tryptophan in the initial startup stage. The appropriate engineering strategy should be further studied to facilitate the actual application of granular sludge for nitrogen removal on a large scale.

Effect of Endometrial Stem Cell-derived Cytokine Cocktail on a Mouse Model of Myocardial Reperfusion Injury.

Objective To study the effect of endometrial stem cells (EnSCs) derived cytokine cocktail (EdCC) on myocardial ischemic reperfusion injury (I/R) in a mouse model. Methods EdCC was concentrated from the culture medium of EnSCs with Millipore ultra-filtration technology and was administrated to a myocardial I/R mouse models through tail vein injection. The infarct area was determined by TTC/Evans Blue staining. The apoptotic cells were counted by TUNEL assay and the protein level of cleaved caspase 3 was evaluated by Western blotting. Results The EdCC extraction efficiency was (222.4±29.3) Μg/10(6) cells in every 24 h,but the protein gradually degraded under-80 ℃ storage. As compared with I/R group,100 Μg fresh EdCC decreased infarct area (P=0.001),reduced apoptotic nuclei in the infarct border (P=0.019),and inhibited cleaved caspase 3 expression (P=0.002). Increasing EdCC dosage did not further reduce the infarct area. The myocardial protective effect of EdCC diminished after 90 days' storage under-80 ℃. Conclusion EdCC reduces myocardial I/R injury through protecting cardiomyocytes from apoptosis within 30 days storage under-80 ℃.

Divergent microbial structure still results in convergent microbial function during arrested anaerobic digestion of food waste at different hydraulic retention times.

In this study, two arrested anaerobic digestion bioreactors, fed with food waste, operated under different hydraulic retention times (HRTs) exhibited similar total volatile fatty acid (VFA) yields (p = 0.09). 16S rRNA gene sequencing revealed distinct microbial structure (p = 0.02) at the two HRTs. However, between the two HRTs, there were no differences in potential (DNA) and extant (mRNA) functionality for the production of acetic (AA)-, propionic (PA)-, butyric (BA)- and valeric-acid (VA), as indicated by the metagenome and metatranscriptome data, respectively. The highest potential and extant functionality for PA production in the reactor microbiomes mirrored the highest abundance of PA in the reactor effluents. Meta-omics analysis of BA production indicated possible metabolite exchange across different community members. Notably, the basis for similar VFA production performance observed under the HRTs tested lies in the community-level redundancy in convergent acidification functions and pathways, rather than trends in community-level structure alone.

Efficient nitrogen removal by microalgal-bacterial granular sludge-marimo coupling process.

Current biological wastewater treatment processes usually have a drawback of insufficient nitrogen (N) removal, contributing to the ubiquitous eutrophication of aquatic ecosystems globally. To address such a challenging situation, this study explored an innovative microalgal-bacterial granular sludge-marimo (MBGS-MA) coupling process. The process removed 83.4 % of N with the effluent N concentration of 4.0 mg/L. With the growth of MBGS, there was a shift towards genes associated with nitrification and denitrification, and away from ammonia assimilation genes, revealing internal mechanism of the shift of N removal pathway. Contrarily, MA could use gaseous N2 with the N fixing genes in MA enriched, and the genes abundance related to assimilatory nitrate reduction were also raised under the mutualistic interactions between Proteobacteria and Cyanobacteria, which was beneficial to achieve efficient N removal. These findings may open a new horizon for developing innovative hybrid microalgal-bacterial processes aimed at high-efficiency N removal from wastewater.

A typical acidic extracellular polysaccharide alludes to algae-bacteria-collaboration in microalgal-bacterial symbiosis.

Microalgal-bacterial symbioses are prevalent in aquatic ecosystems and play a pivotal role in carbon sequestration, significantly contributing to global carbon cycling. The understanding of the contribution of exopolysaccharides (EPSs), a crucial carbon-based component, to the structural integrity of microalgal-bacterial symbioses remains insufficiently elucidated. To address this gap, our study aims to enhance our comprehension of the composition and primary structure of EPSs within a specific type of granular microalgal-bacterial symbiosis named microalgal-bacterial granular sludge (MBGS). Our investigation reveals that the acidic EPSs characteristic of this symbiosis have molecular weights ranging from several hundred thousand to over one million Daltons, including components like glucopyranose, galactopyranose, mannose, and rhamnose. Our elucidation of the backbone linkage of a representative exopolysaccharide revealed a →3)-ß-D-Galp-(1→4)-ß-D-Glcp-(1→ glycosidic linkage. This linear structure closely resembles bacterial xanthan, while the branched chain structure bears similarities to algal EPSs. Our findings highlight the collaborative synthesis of acidic EPSs by both microalgae and bacteria, emphasizing their joint contribution in the production of macromolecules within microalgal-bacterial symbiosis. This collaborative synthesis underscores the intricate molecular interactions contributing to the stability and function of these symbiotic relationships.

Enhancing the performance of microalgal-bacterial systems with sodium bicarbonate: A step forward to carbon neutrality of municipal wastewater treatment.

The microalgal-bacterial granular sludge (MBGS) process, enhanced with sodium bicarbonate (NaHCO3), offers a sustainable alternative for wastewater treatment aiming for carbon neutrality. This study demonstrates that NaHCO3, which can be derived from the flue gases and alkaline textile wastewater, significantly enhances pollutant removal and biomass production. Optimal addition of NaHCO3 was found to achieve an inorganic-to-organic carbon ratio of 1.0 and a total carbon-to-nitrogen ratio of 5.0. Metagenomic analysis and structural equation modeling showed that NaHCO3 addition increased dissolved oxygen concentrations and pH levels, creating a more favorable environment for key microbial communities, including Proteobacteria, Chloroflexi, and Cyanobacteria. Confocal laser scanning microscopy further confirmed enhanced interactions between Cyanobacteria and Proteobacteria/Chloroflexi, facilitating the MBGS process. These microbes harbored functional genes (gap2, GLU, and ppk) critical for removing organics, nitrogen, and phosphorus. Carbon footprint analysis revealed significant reductions in CO2 emissions by the NaHCO3-added MBGS process in representative countries (China, Australia, Canada, Germany, and Morocco), compared to the conventional activated sludge process. These findings highlight the effectiveness of NaHCO3 in optimizing MBGS process, establishing it as a key strategy in achieving carbon-neutral wastewater treatment globally.

Nitrogen removal pathways in lake restoration using microalgal-bacterial granular sludge: Unraveling influence of organics and carbon to nitrogen ratio.

This study investigated the performance of microalgal-bacterial granular sludge (MBGS) in the restoration of Qingling Lake and Huangjia Lake, focusing on nitrogen removal under varying water quality conditions. Significant color changes in MBGS and differences in granule characteristics were observed, with Qingling Lake demonstrating superior removal efficiencies for ammonia nitrogen, nitrate nitrogen, and total nitrogen compared to Huangjia Lake. Stoichiometric analysis revealed that when the chemical oxygen demand (COD) and carbon-to-nitrogen (C/N) ratios were less than 20 mg/L and 20, respectively, assimilatory nitrate reduction was positively correlated with both, whereas denitrification was negatively correlated. Gene function analysis showed that Qingling Lake had a more active microbial community supporting efficient nitrogen metabolism. The findings highlighted the enormous potential of MBGS in lake restoration, demonstrating its ability to adapt to different COD concentrations and C/N ratios by altering its nitrogen removal pathways.

Low-carbon wastewater treatment and resource recovery of recirculating aquaculture system by immobilized chlorella vulgaris based on machine learning optimization.

Immobilized microalgae biotechnologies can conserve water and space by low-carbon wastewater treatment and resource recovery in a recirculating aquaculture system (RAS). However, technical process parameters have been unoptimized considering the mutual interaction between factors. In this study, machine learning optimized the parameters of alginate-immobilized Chlorella vulgaris (C. vulgaris), that is, 474 µmol/(m2·s) of light intensity, 23 × 106 cells/mL for initial cell number, and 2.07 mm particle size. Importantly, under continuous illumination, the immobilized C. vulgaris and microalgal-bacterial consortium improved water purification and biomass reutilization. Transcriptomics of C. vulgaris showed enhanced nitrogen removal by increasing pyridine nucleotide and lipid accumulation via enhanced triacylglycerol synthesis. Symbiotic bacteria upregulated genes for nitrate reduction and organic matter degradation, which stimulated biomass accumulation through CO2 fixation and starch synthesis. The recoverable microalgae (1.94 g/L biomass, 47 % protein, 26.23 % lipids), struvite (64.79 % phosphorus), and alginate (79.52 %) every two weeks demonstrates a low-carbon resource recovery in RAS.

Microbial mechanisms for higher hydrogen production in anaerobic digestion at constant temperature versus gradient heating.

BACKGROUND: Clean energy hydrogen (H2) produced from abundant lignocellulose is an alternative to fossil energy. As an essential influencing factor, there is a lack of comparison between constant temperatures (35, 55 and 65 °C) and gradient heating temperature (35 to 65 °C) on the H2 production regulation potential from lignocellulose-rich straw via high-solid anaerobic digestion (HS-AD). More importantly, the microbial mechanism of temperature regulating H2 accumulation needs to be investigated. RESULTS: Constant 65 °C led to the lowest lignin residue (1.93%) and the maximum release of cellulose and hemicellulose, and the highest H2 production (26.01 mL/g VS). H2 production at 35 and 55 °C was only 14.56 and 24.13 mL/g VS, respectively. In order to further explore the potential of ultra-high temperature (65 °C), HS-AD was performed by gradient heating conditions (35 to 65 °C). However, compared to constant 65 °C, gradient heating conditions led to higher lignin residue (2.49%) and lower H2 production (13.53 mL/g VS) than gradient heating conditions (47.98%). In addition, metagenomic analysis showed the cellulose/hemicellulose hydrolyzing bacteria and genes (mainly Thermoclostridium, and xynA, xynB, abfA, bglB and xynD), H2-producing bacteria and related genes (mainly Thermoclostridium, and nifD, nifH and nifK), and microbial movement and metabolic functions were enriched at 65 °C. However, the enrichment of two-component systems under gradient heating conditions resulted in a lack of highly-enriched ultra-high-temperature cellulose/hemicellulose hydrolyzing genera and related genes but rather enriched H2 consumption genera and genes (mainly Acetivibrio, and hyaB and hyaA) resulting in a weaker H2 production. CONCLUSIONS: The lignin degradation process does not directly determine H2 accumulation, which was actually regulated by bacteria/genes contributing to H2 production/consumption. In addition, it is temperature that enhances the hydrolysis process of lignin rather than lignin-degrading enzymes, bacteria and genes by promoting microbial material transfer and metabolism. In terms of temperature, one of the key parameters of HS-AD for H2 production, we developed an important regulatory strategy, enriched the theoretical basis of temperature regulation for H2 production to further expanded the research horizon in this field. Video Abstract.

Structural basis for an early stage of the photosystem II repair cycle in Chlamydomonas reinhardtii.

Photosystem II (PSII) catalyzes water oxidation and plastoquinone reduction by utilizing light energy. It is highly susceptible to photodamage under high-light conditions and the damaged PSII needs to be restored through a process known as the PSII repair cycle. The detailed molecular mechanism underlying the PSII repair process remains mostly elusive. Here, we report biochemical and structural features of a PSII-repair intermediate complex, likely arrested at an early stage of the PSII repair process in the green alga Chlamydomonas reinhardtii. The complex contains three protein factors associated with a damaged PSII core, namely Thylakoid Enriched Factor 14 (TEF14), Photosystem II Repair Factor 1 (PRF1), and Photosystem II Repair Factor 2 (PRF2). TEF14, PRF1 and PRF2 may facilitate the release of the manganese-stabilizing protein PsbO, disassembly of peripheral light-harvesting complexes from PSII and blockage of the QB site, respectively. Moreover, an α-tocopherol quinone molecule is located adjacent to the heme group of cytochrome b559, potentially fulfilling a photoprotective role by preventing the generation of reactive oxygen species.

Impact of zero-valent iron on nitrifying granular sludge for 17α-ethinylestradiol removal and its mechanism.

Aerobic granulation of nitrifying activated sludge could enhance the removal of 17α-ethinylestradiol (EE2) via abiotic nitration induced by reactive nitrogen species, cometabolism by ammonia-oxidizing bacteria and biodegradation by heterotrophic bacteria. Zero-valent iron (ZVI), a promising and low-cost material, has previously been applied to effectively enhance biological wastewater treatment. The impact and the effect mechanism of ZVI on nitrifying granular sludge (NGS) for EE2 removal was investigated in this study. The results showed that the addition of ZVI achieved better EE2 removal, though ZVI was not conducive to the accumulation of nitrite in NGS which reduced the abiotic transformation of EE2. Moreover, ZVI enriched heterotrophic denitrifying bacteria such as Arenimonas, thus changing the EE2 removal pathway and improving the degradation and mineralization of EE2. In addition, ZVI reduced the emission risk of the greenhouse gas N2O and strengthened the stability of the granules. Metagenomic analysis further revealed that the functional genes related to EE2 mineralization, nitrite oxidation, N2O reduction and quorum sensing in NGS were enriched with ZVI addition. This study provides meaningful guidance for ZVI application in the NGS process to achieve efficient and simultaneous removal of ammonia and emerging contaminants.

Enhancing biolipid production and self-flocculation of Chlorella vulgaris by extracellular polymeric substances from granular sludge with CO2 addition: Microscopic mechanism of microalgae-bacteria symbiosis.

Microalgae-bacteria symbiotic systems were known to have great potential for simultaneous water purification and resource recovery, among them, microalgae-bacteria biofilm/granules have attracted much attention due to its excellent effluent quality and convenient biomass recovery. However, the effect of bacteria with attached-growth mode on microalgae, which has more significance for bioresource utilization, has been historically ignored. Thus, this study attempted to explore the responses of C. vulgaris to extracellular polymeric substances (EPS) extracted from aerobic granular sludge (AGS), for enhancing the understanding of microscopic mechanism of attached microalgae-bacteria symbiosis. Results showed that the performance of C. vulgaris was effectively boosted with AGS-EPS treatment at 12-16 mg TOC/L, highest biomass production (0.32±0.01 g/L), lipid accumulation (44.33±5.69%) and flocculation ability (20.83±0.21%) were achieved. These phenotypes were promoted associated with bioactive microbial metabolites in AGS-EPS (N-acyl-homoserine lactones, humic acid and tryptophan). Furthermore, the addition of CO2 triggered carbon flow into the storage of lipids in C. vulgaris, and the synergistic effect of AGS-EPS and CO2 for improving microalgal flocculation ability was disclosed. Transcriptomic analysis further revealed up-regulation of synthesis pathways for fatty acid and triacylglycerol that was triggered by AGS-EPS. And within the context of CO2 addition, AGS-EPS substantially upregulated the expression of aromatic protein encoding genes, which further enhanced the self-flocculation of C. vulgaris. These findings provide novel insights into the microscopic mechanism of microalgae-bacteria symbiosis, and bring new enlightenment to wastewater valorization and carbon-neutral operation of wastewater treatment plants based on the symbiotic biofilm/biogranules system.