Mining phase separation-related diagnostic biomarkers for endometriosis through WGCNA and multiple machine learning techniques: a retrospective and nomogram study.

OBJECTIVE: The objective of this study was to investigate the role of phase separation-related genes in the development of endometriosis (EMs) and to identify potential characteristic genes associated with the condition. METHODS: We used GEO database data, including 74 non-endometriosis and 74 varying-degree EMs patients. Our approach involved identifying significant gene modules, exploring gene intersections, identifying core genes, and screening for potential EMs biomarkers using weighted gene co-expression network analysis (WGCNA) and various machine learning approaches. We also performed gene set enrichment analysis (GSEA) to understand relevant pathways. This comprehensive approach helps investigate EMs genetics and potential biomarkers. RESULTS: Nine genes were identified at the intersection, suggesting their involvement in EMs. GSEA linked DEGs to pathways like complement and coagulation cascades, DNA replication, chemokines, apical plasma membrane processes, and diseases such as Hepatitis B, Human T-cell leukemia virus 1 infection, and COVID-19. Five feature genes (FOS, CFD, CCNA1, CA4, CST1) were selected by machine learning for an effective EMs diagnostic nomogram. GSEA indicated their roles in mismatch repair, cell cycle regulation, complement and coagulation cascades, and IL-17 inflammation. Notable differences in immune cell proportions (CD4 T cells, CD8 T cells, DCs, macrophages) were observed between normal and disease groups, suggesting immune involvement. CONCLUSIONS: This study suggests the potential involvement of phase separation-related genes in the pathogenesis of endometriosis (EMs) and identifies promising biomarkers for diagnosis. These findings have implications for further research and the development of new therapeutic strategies for EMs.

Analysis of the regulatory mechanism of exogenous IAA-mediated tryptophan accumulation and synthesis of endogenous IAA in Chlorococcum humicola.

The activated sludge method is widely used for the treatment of phenol-containing wastewater, which gives rise to the problem of toxic residual sludge accumulation. Indole-3-acetic acid (IAA), a typical phytohormone, facilitates the microalgal resistance to toxic inhibition while promoting biomass accumulation. In this study, Chlorococcum humicola (C. humicola) was cultured in toxic sludge extract and different concentrations of IAA were used to regulate its physiological properties and enrichment of high value-added products. Ultimately, proteomics analysis was used to reveal the response mechanism of C. humicola to exogenous IAA. The results showed that the IAA concentration of 5 × 10-6 mol/L (M) was most beneficial for C. humicola to cope with the toxic stress in the sludge extract medium, to promote the activity of rubisco enzyme, to enhance the efficiency of photosynthesis, and, finally, to accumulate protein as a percentage of specific dry weight 1.57 times more than that of the control group. Exogenous IAA altered the relative abundance of various amino acids in C. humicola cells, and proteomic analyses showed that exogenous IAA stimulated the algal cells to produce more indole-3-glycerol phosphate (IGP), indole, and serine by up-regulating the enzymes. These precursors are converted to tryptophan under the regulation of tryptophan synthase (A0A383V983), and tryptophan can be metabolized to endogenous IAA to promote the growth of C. humicola. These findings have important implications for the treatment of toxic residual sludge while enriching for high-value amino acids.

Discovering and Validating Cuproptosis-Associated Marker Genes for Accurate Keloid Diagnosis Through Multiple Machine Learning Models.

Background: Keloid is a common condition characterized by abnormal scarring of the skin, affecting a significant number of individuals worldwide. Objective: The occurrence of keloids may be related to the reduction of cell death. Recently, a new cell death mode that relies on copper ions has been discovered. This study aimed to identify novel cuproptosis-related genes that are associated with keloid diagnosis. Methods: We utilized several gene expression datasets, including GSE44270 and GSE145725 as the training group, and GSE7890, GSE92566, and GSE121618 as the testing group. We integrated machine learning models (SVM, RF, GLM, and XGB) to identify 10 cuproptosis-related genes (CRGs) for keloid diagnosis in the training group. The diagnostic capability of the identified CRGs was validated using independent datasets, RT-qPCR, Western blotting, and IHC analysis. Results: Our study successfully categorized keloid samples into two clusters based on the expression of cuproptosis-related genes. Utilizing WGCNA analysis, we identified 110 candidate genes associated with cuproptosis. Subsequent functional enrichment analysis results revealed that these genes may play a regulatory role in cell growth within keloid tissue through the MAPK pathway. By integrating machine learning models, we identified CRGs that can be used for diagnosing keloid. The diagnostic efficacy of CRGs was confirmed using independent datasets, RT-qPCR, Western blotting, and IHC analysis. GSVA analysis indicated that high expression of CRGs influenced the gene set related to ECM receptor interaction. Conclusion: This study identified 10 cuproptosis-related genes that provide insights into the molecular mechanisms underlying keloid development and may have implications for the development of targeted therapies.

Bisphenols in Aquatic Products from South China: Implications for Human Exposure.

In this study, 245 representative samples of aquatic products were selected from local markets in Shenzhen by stochastic sampling. The samples comprised eight species and fell into three aquatic product categories: fish, crustaceans, and bivalves. A total of eight BPs were determined by liquid chromatography coupled with mass spectrometry, namely, bisphenol A (BPA), bisphenol AF (BPAF), bisphenol AP (BPAP), bisphenol B (BPB), bisphenol S (BPS), bisphenol P (BPP), bisphenol Z (BPZ), and bisphenol F (BPF). All BPs were detected in aquatic products, except for BPAF, indicating pervasive contamination by BPs in aquatic products. BPS demonstrated the highest detection rate both before and after enzymatic hydrolysis, whereas BPAP exhibited the lowest detection rate before enzymatic hydrolysis and BPB displayed the lowest detection rate after enzymatic hydrolysis. The concentration difference before and after enzymatic hydrolysis proved to be statistically significant. Moreover, 49-96% of BPs in aquatic products were found in the combined state, underscoring the essentiality of conducting detections on aquatic product samples following enzymatic hydrolysis. While the health risks associated with ingesting BPs residues through aquatic product consumption were found to be minimal for residents at risk of exposure, the results suggest the necessity for more stringent regulations governing the consumption of aquatic products.

Proteomics reveals toxin tolerance and polysaccharide accumulation in Chlorococcum humicola under high CO2 concentration.

Algae have great application prospects in excess sludge reclamation and recovery of high-value biomass. Chlorococcum humicola was cultivated in this research, using sludge extract (mixed with SE medium) with additions of 10%, 20%, and 30% CO2 (v/v). Results showed that under 20% CO2, the dry weight and polysaccharide yield reached 1.389 ± 0.070 g/L and 313.49 ± 10.77 mg/L, respectively. 10% and 20% CO2 promoted the production of cellular antioxidant molecules to resist the toxic stress and the toxicity of 20% CO2 group decreased from 62.16 ± 3.11% to 33.02 ± 3.76%. 10% and 20% CO2 accelerated the electron transfer, enhanced carbon assimilation, and promoted the photosynthetic efficiency, while 30% CO2 led to photosystem damage and disorder of antioxidant system. Proteomic analysis showed that 20% CO2 mainly affected energy metabolism and the oxidative stress level on the early stage (10 d), while affected photosynthesis and organic substance metabolism on the stable stage (30 d). The up-regulation of PSII photosynthetic protein subunit 8 (PsbA, PsbO), A0A383W1S5 and A0A383VRI4 promoted the efficiency of PSII and chlorophyll synthesis, and the up-regulation of A0A383WH74 and A0A2Z4THB7 led to the accumulation of polysaccharides. The up-regulation of A0A383VDH1, A0A383VX37 and A0A383VA86 promoted respiration. Collectively, this work discloses the regulatory mechanism of high-concentration CO2 on Chlorococcum humicola to overcome toxicity and accumulate polysaccharides.

The efficient capture of polysaccharides in Tetradesmus obliquus of indole-3-acetic acid coupling sludge extraction.

Polysaccharide is an important biomass of algae. The sludge extract is rich in organic substances, which can be used by algae for biomass growth and high-value biomass synthesis, but its organic toxicity has an inhibitory effect on algae. To overcome inhibition and improve polysaccharide enrichment, Tetradesmus obliquus was cultured with sludge extract with different indole-3-acetic acid (IAA) concentrations. Within 30 days of the culture cycle, T. obliquus showed in good condition at the IAA dosage content of 10-6 M, the maximum cell density and dry weight were respectively (106.78 ± 2.20) × 106 cell/mL and 2.941 ± 0.067 g/L while the contents of chlorophyll-a, chlorophyll-b, and carotenoid were 1.79, 1.91 and 2.80 times that of the blank group, respectively. The highest polysaccharide accumulation was obtained under this culture condition, reaching 533.15 ± 21.11 mg/L on the 30th day, which was 2.49 times that in the blank group. By FT-IR and NMR analysis, it was found that the polysaccharides of T. obliquus were sulfated polysaccharide with glucose and rhamnose as the main monosaccharides. Proteomic showed that the up-regulation of A0A383WL26 and A0A383WLM8 enhanced the light trapping ability, and A0A383WMJ2 enhanced the accumulation of NADPH. The up-regulation of A0A383WHD5 and A0A383WAY6 indicated that IAA culture could repair the damage caused by sludge toxicity, thus promoting the accumulation of biomass. The above findings provided new insights into the mechanism of sludge toxicity removal of T. obliquus and the enhancement of the polysaccharide accumulation effect under different concentrations of IAA.

Excessive composite pollution carbon sources enhance the bio-fertilizer efficiency of Tetradesmus obliquus: focused on cultivation period.

Microalgae can use carbon sources in sludge extract prepared from sludge. Moreover, the high concentration of CO2 and the large number of carbon sources in the liquid phase will promote microalgae growth and metabolism. In this experiment, Tetradesmus obliquus was cultivated with sludge extract at 30% CO2. Algae liquid (the name used to describe the fertilizer made in this research) was further prepared as lettuce fertilizer. The effect of different times of microalgae culture (10, 15, 20, 25, and 30 days) on the fertilizer efficiency of the algae liquid was evaluated by lettuce hydroponic experiments. The findings indicate that lettuce cultivated in algae liquid collected on the 15th and 30th days exhibited superior performance in terms of growth, antioxidant capacity, and nutritional quality. We analyzed the experimental results in the context of microalgae metabolic mechanisms, aiming to contribute experience and data essential for the development of industrial microalgae fertilizer production.

Indole-3-acetic acid mediated removal of sludge toxicity by microalgae: Focus on the role of extracellular polymeric substances.

The use of indole-3-acid (IAA) as an additive aided in achieving the objectives of reducing sludge extract toxicity, increasing Tetradesmus obliquus biomass yield, and enhancing extracellular polysaccharide production. Proteomics analysis can unveil the microalgae's response mechanism to sludge toxicity stress. With 10-6 M IAA addition, microalgae biomass reached 3.426 ± 0.067 g/L. Sludge extract demonstrated 78.3 ± 3.2% total organic carbon removal and 72.2 ± 2.1% toxicity removal. Extracellular polysaccharides and proteins witnessed 2.08 and 1.76-fold increments, respectively. Proteomic analysis indicated that Tetradesmus obliquus directed carbon sources towards glycogen accumulation and amino acid synthesis, regulating pathways associated with carbon metabolism (glycolysis, TCA cycle, and amino acid metabolism) to adapt to the stressful environment. These findings lay the groundwork for future waste sludge treatment and offer novel insights into microalgae cultivation and extracellular polysaccharide enrichment in sludge.

Analysis of the mechanism of exogenous indole-3-acetic acid on the enrichment of d-glucose in Chlorococcum humicola cultured by sludge extracts.

Addressing problems of high organic toxicity in the wastewater treatment process, microalgae have been used to reduce the toxicity in sludge and to synthesize non-toxic and recoverable biomass of resources. Phytohormone is a core regulator of plant growth and current research has generally focused on their promotion of cell division and cell expansion. Effects of phytohormone on the enrichment mechanism of microalgae directional polysaccharides accumulation remain poorly elucidated. This study was carried out to investigate the effects of exogenous indole-3-acetic acid (IAA) on growth characteristics, biomass accumulation, and photosynthesis capacity of Chlorococcum humicola cultured in sludge extract and further find the d-glucose enrichment mechanism of it through proteomic. The results indicated that the optimal culture conditions were the 75 % sludge extract and 25 % selenite enrichment (SE) medium with 5 × 10-6 mol/L indole-3-acetic acid. Polysaccharides increased significantly from day 20 and accumulated to (326.59 ± 13.06) mg/L on day 30, in which the d-glucose proportion increased to 61.53 %. Most notably, proteomic tests were performed and found that the photosynthesis-related proteins including the differential proteins of photosystem electron transport, ATP and NADPH catalytic synthesis were significantly up-regulated. At the end of the path, three pathways of d-glucose enrichment with α-d-Glucose-1P as a precursor were summarized through indole-3-acetic acid activation on amylase, endoglucanase and Beta-glucosidase, etc. These results provide insights to explore the directed enrichment of biomass in Chlorococcum humicola by indole-3-acetic acid.

Regulatory mechanism of high-concentration CO2 on polysaccharide accumulation in Tetradesmus obliquus cultured in sludge extract.

Microalgae such as Tetradesmus obliquus have great potential in immobilizing high-concentration CO2 and removing highly toxic organic matters, which could be produced from coal chemical industry and coal chemical wastewater biological treatment process. In this study, Tetradesmus obliquus was cultured in sludge extract and high-concentration CO2 was added. The maximum cell density and dry weight were respectively (111.46 ± 4.87) × 106 cell/mL and 3.365 ± 0.168 g/L under 30% CO2. Tetradesmus obliquus accumulated the most polysaccharides (629.60 ± 31.48 mg/L) on the 30th day under 30% CO2. The results of proteomic showed that the upregulation of A0A2Z4THB7 and A0A383VAT1 promoted polysaccharide accumulation. Polysaccharide was mainly formed at the stable phase instead of the log-growth phase due to the abiotic stress caused by high TOC at the log-growth phase. Collectively, this study revealed the regulatory mechanism of high-concentration CO2 on the toxicity removal and accumulation of polysaccharides in Tetradesmus obliquus.

Improvement of coal gasification reverse osmosis concentrate treatment by Cu-Co-Mn/AC catalytic ozonation.

Approximately 20% of concentrate will be produced from coal gasification wastewater after reverse osmosis treatment. The organic matter contained in the concentrate affects its evaporation crystallisation; therefore, the refractory organics must be removed. In this study, Cu-Co-Mn/AC catalytic ozonation was used to treat reverse osmosis concentrate (ROC). With the addition of the Cu-Co-Mn/AC catalyst, the production of ·OH increased by 82 µmol/L, thereby enhancing the ozonation performance. The pH, ozone dosage, and catalyst dosage all affected the catalytic ozonation performance. By constructing a response surface model, it was found that the catalyst dosage had the most significant effect on the catalytic ozonation performance. The predicted optimal reaction conditions were pH = 9.02, ozone dosage = 1.08 g/L, and catalyst dosage = 1.33 g/L, under which the chemical oxygen demand (COD) removal reached a maximum of 81.49%.

Analysis of photosynthetic pigments pathway produced by CO2-toxicity-induced Scenedesmus obliquus.

The coal-to-gas process produces carbon dioxide, which increases global warming, and its wastewater treatment generates sludge with high organic toxicity. Scenedesmus obliquus is a potential solution to such environmental problems, and photosynthetic pigments are the focus of this study. The optimal concentration of CO2 for the growth of Scenedesmus obliquus was found to be 30 % after increasing the concentration of CO2 (0.05 %-100 %). The accumulation of photosynthetic pigments during cultivation could reach 31.74 ± 1.33 mg/L, 11.21 ± 0.42 mg/L, and 5.59 ± 0.19 mg/L respectively, and the organic toxicity of sludge extract could be reduced by 44.97 %. Upregulation of A0A383VSL5, A0A383WMQ3, and A0A2Z4THB7 as photo systemic oxygen release proteins and propylene phosphate isomerase resulted in oxygen-evolving proteins in photosystem II, electron transport in photosystem I, and intermediates in carbon fixation. This is achieved by increasing the intracellular antennae protein and carbon fixation pathway, allowing Scenedesmus obliquus to both tolerate and fix CO2 and reduce the organic toxicity of sludge. These findings provide insights into the innovative strategy underlining the fixation of CO2, treatment and disposal of industrial residual sludge, and the enhancement of microalgal biomass production.

Self-regulation mechanism difference of Chlorella vulgaris and Scenedesmus obliquus in toxic sludge extract caused by hydroquinone biodegradation.

Chlorella vulgaris (C. vulgaris) and Scenedesmus obliquus (S. obliquus) were compared to remove toxicity under conditions of sludge extract cultivation for 30 days. The toxicity of sludge extract, the growth characteristics, photosynthetic pigment, superoxide dismutase (SOD) enzyme and catalase (CAT) enzyme activities of the two microalgae were studied by contrast. The results showed that small molecular organic matter (<500 Da) was more easily utilized by microalgae. The toxicity in the toxic group of C. vulgaris and S. obliquus on the 30th day decreased to 56.8 ± 1.2% and 60.7 ± 2.8%, respectively. In the toxic group, the maximal SOD enzyme activity of C. vulgaris and S. obliquus were 2.02 U/mg proteins and 8.21 U/mg proteins, respectively, demonstrating that toxicity caused more oxidative damage to S. obliquus than to C. vulgaris. Proteomics analysis revealed that C. vulgaris mainly regulates energy synthesis and distribution primarily through sugar metabolism, and biomass synthesis primarily through carbon metabolism, whereas S. obliquus mainly regulates energy synthesis and distribution primarily through sugar metabolism and oxidative phosphorylation, resulting in sludge toxicity stress regulation.

Effects of different concentrations of CO2 on Scenedesmus obliquus to overcome sludge extract toxicity and accumulate biomass.

Large amounts of toxic excess sludge as well as high concentrations of carbon dioxide can be produced in coal-gasification industry. Microalgae has huge potential in the use of nutrients, the removal of toxic organic matter in excess sludge and CO2 fixation. At the same time, the cultivation of the microalgae and the accumulation of high-quality biomass are also the key problems of concern. In this study, the growth and biomass synthesis of Scenedesmus obliquus cultured in sludge extract under 0%-15% (v/v) CO2 were investigated. Results indicated that the highest microalgae biomass yield of 1.609 ± 0.012 g/L can be achieved under 15% CO2 on the 30th day. The maximal photochemical efficiency of PSⅡ (Fv/Fm) decreased in the first 12 h and then increased with the culture time, and the decline amplitude decreased with the increase of the CO2 concentration, indicating that CO2 slowed down the toxic inhibition of sludge extract to Scenedesmus obliquus, which was expressed as the down-regulation of p53 signaling pathway and protein A0A383WFI7. Proteomic analysis showed that under high-concentration CO2, the protein interaction network with the protein of photosystem II assembly (A0A383VSL5) as the core protein regulated the growth of Scenedesmus obliquus in terms of energy metabolism and material transportation. On the 4th day, Methyltransf_11 domain-containing protein (A0A383VH03) was up-regulated and promoted lipid synthesis, leading to the accumulation of lipids in Scenedesmus obliquus in the early stage and the increase of polysaccharides in the later stage. Collectively, this study revealed the regulation mechanism of CO2 on toxicity removal and carbon distribution of Scenedesmus obliquus.

Bacterial diversity evolution process based on physicochemical characteristics of sludge treating hydroquinone during acclimation.

Hydroquinone is one of the main pollutants in coal-gasification wastewater, which is biologically toxic and difficult to remove. The aerobic biodegradation rate, organic toxicity, and microbial community structure at different acclimation stages of degradation of hydroquinone by activated sludge were investigated. In each acclimation cycle, the removal of hydroquinone reached 100% after 5 days, indicating that high-concentration hydroquinone in the activated sludge could be completely biodegraded. When the microbial flora was inhibited by the influent hydroquinone, the enzyme system experienced stress conditions and led to the secretion of secondary metabolites, extracellular protein of 5-10 kDa mainly contributing to the sludge organic toxicity. Microbial diversity analysis showed that with the increase of the concentration of hydroquinone, ß-Proteus bacteria such as Azoarcus and Dechloromonas gradually accumulated, which improved the removal of hydroquinone with aerobic activated sludge in the sequencing batch reactor (SBR) system. As the inhibition degree exceeded the appropriate tolerance range of microorganisms, bacteria would secrete much more secondary metabolites, and the organic toxicity of sludge would reach a relatively high level.

Study on the differences in sludge toxicity and microbial community structure caused by catechol, resorcinol and hydroquinone with metagenomic analysis.

The aerobic biodegradation rate, organic toxicity and microbial community structure of activated sludge acclimated by catechol, resorcinol and hydroquinone were investigated, to study the relationship between microbial structure and sludge organic toxicity caused by phenolic compounds. At the stable operation stage, the degradation rates of the dihydroxy benzenes in a single sequencing batch reactor (SBR) cycle were followed the order: resorcinol (89.71%) > hydroquinone (85.64%) > catechol (59.62%). Sludge toxicity bioassay indicated that the toxicity of sludge was catechol (45.63%) > hydroquinone (40.28%) > resorcinol (38.15%). The accumulation of secondary metabolites such as 5-10 kDa tryptophan and tyrosine protein substances caused the differential sludge toxicity. Microbial metagenomic analysis showed that the toxicity of sludge was significantly related to the microbial community structure. Thauera, Azoarcus, Pseudomonas and other Proteobacteria formed in the sludge during acclimation. Catechol group had the least dominant bacteria and loop ring opening enzyme genes (catA, dmpB, dxnF, hapD) numbers. Therefore, the degradation of catechol was the most difficult than resorcinol and hydroquinone, resulting the highest sludge toxicity.

Latent Epstein-Barr virus infection status and prognosis in patients with newly diagnosed Hodgkin lymphoma in Southeast China: a single-center retrospective study.

PURPOSE: This study aimed to investigate the latent Epstein-Barr virus (EBV) infection status of patients with newly diagnosed Hodgkin lymphoma (HL) and to discuss the relationship between tumor cell EBV status and the prognosis of HL patients. PATIENTS AND METHODS: A total of 134 previously untreated HL patients were analyzed in the study. Epstein-Barr virus encoded RNAs (EBERs) in situ hybridization was performed to detect the EBV status of tumor cells. RESULTS: EBV positive status correlated with sex (p=0.046) and the proportion of extranodal lesions(p=0.037). There was no obvious correlation between EBV status and overall survival (OS) or failure-free survival (FFS) in all cases, but in cases over 50 years old, EBV positive group had an inferior 5-year FFS compared with EBV negative group (38.5%±13.5% vs 90.9%±8.7%, p=0.012). In FFS multivariate analysis of this age subgroup, EBV positive status was associated with significantly inferior survival (HR, 10.10; 95% CI, 1.26-81.08; p=0.030). CONCLUSION: This study demonstrates positive tumor cell EBV status is an unfavorable prognostic factor in elder HL patients.

Deciphering and Suppressing Over-Oxidized Nitrogen in Nickel-Catalyzed Urea Electrolysis.

Urea electrolysis is a prospective technology for simultaneous H2 production and nitrogen suppression in the process of water being used for energy production. Its sustainability is currently founded on innocuous N2 products; however, we discovered that prevalent nickel-based catalysts could generally over-oxidize urea into NO2 - products with ≈80 % Faradaic efficiencies, posing potential secondary hazards to the environment. Trace amounts of over-oxidized NO3 - and N2 O were also detected. Using 15 N isotopes and urea analogues, we derived a nitrogen-fate network involving a NO2 - -formation pathway via OH- -assisted C-N cleavage and two N2 -formation pathways via intra- and intermolecular coupling. DFT calculations confirmed that C-N cleavage is energetically more favorable. Inspired by the mechanism, a polyaniline-coating strategy was developed to locally enrich urea for increasing N2 production by a factor of two. These findings provide complementary insights into the nitrogen fate in water-energy nexus systems.

Comparison of nitrogen removal efficiency and microbial characteristics of modified two-stage A/O, A2/O and SBR processes.

The low temperature of sewage in north China results in low performance of biological treatment at municipal wastewater treatment plants (MWTPs), especially in biological nitrogen removal. A modified two-stage A/O process with an embedded biofilm was proposed to enhance nitrogen removal. The operation performance of a pilot test was compared with an A2/O and SBR process at existing MWTPs to investigate the resistance to low temperature. The microbial communities for the three processes were compared based on the metagenomics results of 16sDNA high-throughput sequencing from activated sludge. The modified A/O resulted in a higher average removal of COD (90.12%) than A2/O (85.23%) and SBR (83.03%), especially of small-molecule organic compounds (< 500 Da) and macromolecular refractory organics (> 5 k Da); the TN removal rate of A2/O, SBR and the modified A/O was also increased from 74.47%, 70.63% and 78.46%, respectively. High-throughput sequencing revealed increased microbial diversity and an abundance of denitrifying functional bacteria was observed in the modified A/O process at low temperatures. The abundance of nitrite oxidation bacteria (NOB) including Nitrosomonas and Nitrospira, the amount was 1.76% and 2.34% in modified A/O, respectively, whereas NOB only accounted for 1.82% in A2/O and 1.35% in SBR.

Cultivation of energy microalga Chlorella vulgaris with low-toxic sludge extract.

Chlorella vulgaris was cultivated in different proportions of activated sludge extracts, which was from the treatment of synthetic wastewater containing tetrachlorophenol. The growth period of C. vulgaris could be shortened for about 10 days when sludge extract was mixed into BG11 culture substrate, and the growth of C. vulgaris was promoted during the period of adaptation and logarithmic period. In the stable and decay period, when the proportion of sludge extract increased to 50%, cell proliferation was inhibited. There was an evident positive correlation between the total and average amount of starch polysaccharide with sludge concentration. When C. vulgaris was cultivated with pure sludge extracts, the total amount of starch and polysaccharide was up to 103 and 125 mg/L. Therefore, the low-toxic sludge extracts were more beneficial to the accumulation of carbohydrates. In the 100% sludge extracts culture medium, chlorophyll-a in C. vulgaris was accumulated to 30.2 mg/L on the 25th day. Through the analysis of algal cells' ultrastructures, it was shown that the photosynthesis was strengthened greatly with low-toxic sludge extracts. The results show that the rich heterotrophic carbon source in the sludge extract can be used as an excellent medium for Chlorella. It provides new ideas for the harmless utilization of surplus sludge as a resource. At the same time, the use of nutrients in the sludge extract to cultivate Chlorella is of great significance to low-cost algae cultivation.