Supercritical Carbon Dioxide Extraction of Four Medicinal Mediterranean Plants: Investigation of Chemical Composition and Antioxidant Activity.

With everyday advances in the field of pharmaceuticals, medicinal plants have high priority regarding the introduction of novel synthetic compounds by the usage of environmentally friendly extraction technologies. Herein, a supercritical CO2 extraction method was implemented in the analysis of four plants (chamomile, St. John's wort, yarrow, and curry plant) after which the non-targeted analysis of the chemical composition, phenolic content, and antioxidant activity was evaluated. The extraction yield was the highest for the chamomile (5%), while moderate yields were obtained for the other three plants. The chemical composition analyzed by gas chromatography-high-resolution mass spectrometry (GC-HRMS) and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) demonstrated extraction of diverse compounds including terpenes and terpenoids, fatty acids, flavonoids and coumarins, functionalized phytosterols, and polyphenols. Voltammetry of microfilm immobilized on a glassy carbon electrode using square-wave voltammetry (SWV) was applied in the analysis of extracts. It was found that antioxidant activity obtained by SWV correlates well to 1,1-diphenyl-2-picrylhidrazine (DPPH) radical assay (R2 = 0.818) and ferric reducing antioxidant power (FRAP) assay (R2 = 0.640), but not to the total phenolic content (R2 = 0.092). Effective results were obtained in terms of activity showing the potential usage of supercritical CO2 extraction to acquire bioactive compounds of interest.

Preparation and characterization of lutein ester-loaded oleogels developed by monostearin and sunflower oil.

The marigold (Tagetes erecta L.) flower is rich in lutein ester with many health-promoting activities. In this study, the effects of vegetable oil type and extracting the temperature on the extraction efficiency of lutein ester in the marigold flower were evaluated. Then, the structuring of the lutein ester-loaded sunflower oil with the addition of different amounts of monostearin and cooling temperatures (4 and 20°C) was investigated. The XRD analysis suggested that these oleogels were stabilized by the network formed by monostearin crystals in the sunflower oil. The textural properties (firmness, cohesiveness, and hardness) of oleogels were positively related to the monostearin dosage, but negatively related to the cooling temperature. According to the rheological results, the oleogels belonged to the pseudoplastic gel and their gelation temperature (Tg ) was only related to the concentration of monostearin. The light stability of lutein ester in the oleogels was also significantly improved in a monostearin dosage-dependent manner. PRACTICAL APPLICATIONS: The edible lutein ester-loaded oleogel for foods developed by structuring the sunflower oil with monostearin is introduced in this study. Its texture and rheological properties can be adjusted to cater to different requirements in the food industry by changing the monostearin dosage and cooling temperature. This study provides a reference for the development of other liposoluble nutraceuticals.

Enhanced and Balanced Microalgal Wastewater Treatment (COD, N, and P) by Interval Inoculation of Activated Sludge.

Although chemical oxygen demand (COD) is an important issue for wastewater treatment, COD reduction with microalgae has been less studied compared to nitrogen or phosphorus removal. COD removal is not efficient in conventional wastewater treatment using microalgae, because the algae release organic compounds, thereby finally increasing the COD level. This study focused on enhancing COD removal and meeting the effluent standard for discharge by optimizing sludge inoculation timing, which was an important factor in forming a desirable algae/bacteria consortium for more efficient COD removal and higher biomass productivity. Activated sludge has been added to reduce COD in many studies, but its inoculation was done at the start of cultivation. However, when the sludge was added after 3 days of cultivation, at which point the COD concentration started to increase again, the algal growth and biomass productivity were higher than those of the initial sludge inoculation and control (without sludge). Algal and bacterial cell numbers measured by qPCR were also higher with sludge inoculation at 3 days later. In a semi-continuous cultivation system, a hydraulic retention time of 5 days with sludge inoculation resulted in the highest biomass productivity and N/P removal. This study achieved a further improved COD removal than the conventional microalgal wastewater treatment, by introducing bacteria in activated sludge at optimized timing.

Extraction and Isolation of Natural Organic Compounds from Plant Leaves Using Ionic Liquids.

Plants contain many kinds of natural organic compounds, and their compounds possess many useful properties. Natural organic compounds are important for the development of medicines, pesticides, fragrances, cosmetics, and synthetic chemicals. In this chapter, we introduce efficient methods for extraction and isolation of valuable natural organic compounds from various plant leaves by using cellulose-dissolving ionic liquids. High-polarity ionic liquids, which can dissolve cellulose, contribute to the extraction of natural organic compounds from plant leaves probably by breaking down plant cell walls, which are composed of cellulose, hemicellulose, and lignin. Extraction and isolation of shikimic acid from ginkgo leaves, caffeoylquinic acids from sweet potato leaves, and neral and geranial (which combine to form citral) from lemon myrtle leaves were performed. Ionic liquids can achieve extraction rates greater than those achieved with water and other organic solvents. Graphical Abstract.

What are the effects of soil treatment procedures (sterilization by γ-irradiation and solvent-assisted spiking) on DDE bioaccumulation by earthworms?

Ionizing γ-irradiation and solvent-assisted spiking are frequently applied to eliminate microbial activity and to induce hydrophobic organic compounds (HOCs) into soil, respectively, when studying the accumulation of chemicals in terrestrial organisms. However, the side-effects that may arise from these treatments on soil-HOC interaction and, subsequently, the kinetics and extents of bioaccumulation are not thoroughly understood. To this end, the accumulation of 1,1-dichloro-2,2-bis(p-chlorophenyl)etylene (p,p'-DDE) by Eisenia andrei was studied in sterilized or unsterilized and freshly spiked (FS) or historically contaminated (HC) soils in parallel with an analysis of aliphatic and hydrophilic soil organic matter (SOM) moieties using mid-infrared diffuse reflectance spectroscopy (DRIFT-S). Irradiation did not impart significant changes on spectral SOM descriptors. In contrast, earthworm inhabitation increased the relative presence of aliphatic moieties to a greater extent than hydrophilic ones, reaching or exceeding pre-treatment levels. Overall, effects on SOM chemistry can be ranked as earthworms > spiking > irradiation. Corresponding changes at the bioaccumulation level were observed for the FS soil (i.e., a 27% reduction in bioaccumulation upon sterilization) but not for the HC soil. This implies that in contrast to the interactions between aged p,p'-DDE and sterilized HC soil, the interactions established between freshly added p,p'-DDE and sterilized FS soil were altered by γ-irradiation-induced secondary effects alone or in combination with earthworm inhabitation. Thus, although the soil treatment processes studied here should not drastically impact compound bioaccumulation, they should be considered in mechanistic studies where the qualitative and quantitative aspects of compound-soil (organic matter)-earthworm interactions are at the centre of attention.

Phosphorous-enriched knitting aryl network polymer for the rapid and effective adsorption of aromatic compounds.

A phosphorous-enriched knitting aryl network polymer (named as Ph-PPh3-KAP) was fabricated by one-step crosslinking between triphenylphosphine and benzene, with formaldehyde dimethyl acetal as an external crosslinker. The Ph-PPh3-KAP had a large surface area and good physicochemical stability. Its adsorption performance for aromatic organic compounds was evaluated by using some dyes and benzene ring-containing compounds as the model adsorbates. The results exhibited that it had a rapid and effective adsorption for the aromatic organic compounds due to the hydrogen bonding and polar interactions of the Ph-PPh3-KAP with the target compounds. Then, the Ph-PPh3-KAP was explored as the adsorbent for the solid phase extraction of some phenylurea pesticides from lake water, tomato and cucumber samples prior to HPLC analysis. Under optimal conditions, the linear responses of the phenylurea pesticides were 0.1-100 ng mL-1 for lake water and 0.5-100 ng g-1 for cucumber and tomato samples. The limits of detection for the analytes at S/N = 3 were 0.01-0.02 ng mL-1 for lake water and 0.03-0.05 ng g-1 for cucumber and tomato samples. The recoveries were in the range from 80.8% to 118%. The Ph-PPh3-KAP exhibited a great application potential for extraction of aromatic compounds.

Removal of organic matter of electrodialysis reversal brine from a petroleum refinery wastewater reclamation plant by UV and UV/H202 process.

Direct (UV) and hydrogen peroxide-assisted (UV/H2O2) photolysis were investigated in bench-scale for removing the organic compounds present in the electrodialysis reversal (EDR) brine from a refinery wastewater reclamation plant. In the UV/H2O2 experiments, a COD:H2O2 molar ratios of 1:1, 1:2 and 1:3 were tested by recirculating the brine in the UV reactor for 120 min. Results showed a significant reduction in UVA254, whereas no reduction was observed for chemical oxygen demand (COD), in the UV process, suggesting great cleavage but limited mineralization of the organic matter. UV/H2O2 with C:H2O2 ratio of 1:3 exhibited high efficiency in removing the organic matter (COD removal of 92% with an electrical energy per removal order (EEO) value of 22 kW h m-3). Although the EDR brine has high salinity, no strong scavenging effect of •OH was found in the water matrix due to the high concentration of anions, especially chloride and bicarbonate. Finally, UV/H2O2 with C:H2O2 ratio of 1:3 and residence time of 120 min is an efficient alternative for organic matter removal of EDR brine from refinery wastewater reclamation plant showing total capital cost (CapEx) estimated at US$ 369,653.00 and total operational cost (OpEx), at US$ 1.772 per cubic meter of effluent.

Potential use of membrane bioreactor to treat petroleum refinery effluent: comprehension of dynamic of organic matter removal, fouling characteristics and membrane lifetime.

This study aims to evaluate membrane bioreactor (MBR) performance in a pilot scale to treat petroleum refinery effluent, and has been primarily focused on (1) investigation of dynamics of organic matter removal; (2) characterization of membrane fouling under real hazardous events; (3) evaluation of the effect of fouling on membrane lifetime; and (4) estimate the membrane lifetime. The results have shown that the MBR was able to effectively reduce COD, NH3-N, turbidity, color, phenol and toxicity, and bring them to the levels required to meet disposal and non-potable water reuse standards. The FTIR results showed that organic matter was removed by biological oxidation and/or retained by adsorption in the biological sludge, or retention in the UF membrane, and that SMP was produced during the treatment. In terms of membrane permeability, the results showed that soluble fraction of mixed liquor contributed significantly to membrane fouling. And finally, considering the concept of lifetime based on permeability decline, a membrane lifetime of 7 years is expected.

Managing peatland vegetation for drinking water treatment.

Peatland ecosystem services include drinking water provision, flood mitigation, habitat provision and carbon sequestration. Dissolved organic carbon (DOC) removal is a key treatment process for the supply of potable water downstream from peat-dominated catchments. A transition from peat-forming Sphagnum moss to vascular plants has been observed in peatlands degraded by (a) land management, (b) atmospheric deposition and (c) climate change. Here within we show that the presence of vascular plants with higher annual above-ground biomass production leads to a seasonal addition of labile plant material into the peatland ecosystem as litter recalcitrance is lower. The net effect will be a smaller litter carbon pool due to higher rates of decomposition, and a greater seasonal pattern of DOC flux. Conventional water treatment involving coagulation-flocculation-sedimentation may be impeded by vascular plant-derived DOC. It has been shown that vascular plant-derived DOC is more difficult to remove via these methods than DOC derived from Sphagnum, whilst also being less susceptible to microbial mineralisation before reaching the treatment works. These results provide evidence that practices aimed at re-establishing Sphagnum moss on degraded peatlands could reduce costs and improve efficacy at water treatment works, offering an alternative to 'end-of-pipe' solutions through management of ecosystem service provision.

Biotransformation of trace organic compounds by activated sludge from a biological nutrient removal treatment system.

The removal of trace organic compounds (TOrCs) and their biotransformation rates, kb (LgSS(-)(1)h(-)(1)) was investigated across different redox zones in a biological nutrient removal (BNR) system using an OECD batch test. Biodegradation kinetics of fourteen TOrCs with initial concentration of 1-36µgL(-)(1) in activated sludge were monitored over the course of 24h. Degradation kinetic behavior for the TOrCs fell into four groupings: Group 1 (atenolol) was biotransformed (0.018-0.22LgSS(-)(1)h(-)(1)) under anaerobic, anoxic, and aerobic conditions. Group 2 (meprobamate and trimethoprim) biotransformed (0.01-0.21LgSS(-)(1)h(-)(1)) under anoxic and aerobic conditions, Group 3 (DEET, gemfibrozil and triclosan) only biotransformed (0.034-0.26LgSS(-)(1)h(-)(1)) under aerobic conditions, and Group 4 (carbamazepine, primidone, sucralose and TCEP) exhibited little to no biotransformation (<0.001LgSS(-)(1)h(-)(1)) under any redox conditions. BNR treatment did not provide a barrier against Group 4 compounds.

Combination of granular activated carbon adsorption and deep-bed filtration as a single advanced wastewater treatment step for organic micropollutant and phosphorus removal.

Adsorption onto granular activated carbon (GAC) is an established technology in water and advanced wastewater treatment for the removal of organic substances from the liquid phase. Besides adsorption, the removal of particulate matter by filtration and biodegradation of organic substances in GAC contactors has frequently been reported. The application of GAC as both adsorbent for organic micropollutant (OMP) removal and filter medium for solids retention in tertiary wastewater filtration represents an energy- and space saving option, but has rarely been considered because high dissolved organic carbon (DOC) and suspended solids concentrations in the influent of the GAC adsorber put a significant burden on this integrated treatment step and might result in frequent backwashing and unsatisfactory filtration efficiency. This pilot-scale study investigates the combination of GAC adsorption and deep-bed filtration with coagulation as a single advanced treatment step for simultaneous removal of OMPs and phosphorus from secondary effluent. GAC was assessed as upper filter layer in dual-media downflow filtration and as mono-media upflow filter with regard to filtration performance and OMP removal. Both filtration concepts effectively removed suspended solids and phosphorus, achieving effluent concentrations of 0.1 mg/L TP and 1 mg/L TSS, respectively. Analysis of grain size distribution and head loss within the filter bed showed that considerable head loss occurred in the topmost filter layer in downflow filtration, indicating that most particles do not penetrate deeply into the filter bed. Upflow filtration exhibited substantially lower head loss and effective utilization of the whole filter bed. Well-adsorbing OMPs (e.g. benzotriazole, carbamazepine) were removed by >80% up to throughputs of 8000-10,000 bed volumes (BV), whereas weakly to medium adsorbing OMPs (e.g. primidone, sulfamethoxazole) showed removals <80% at <5,000 BV. In addition, breakthrough behavior was also determined for gabapentin, an anticonvulsant drug recently detected in drinking water resources for which suitable removal technologies are still largely unknown. Gabapentin showed poor adsorptive removal, resulting in rapid concentration increases. Whereas previous studies classified gabapentin as not readily biodegradable, sustained removal was observed after prolonged operation and points at biological elimination of gabapentin within the GAC filter. The application of GAC as filter medium was compared to direct addition of powdered activated carbon (PAC) to deep-bed filtration as a direct process alternative. Both options yielded comparable OMP removals for most compounds at similar carbon usage rates, but GAC achieved considerably higher removals for biodegradable OMPs. Based on the results, the application of GAC in combination with coagulation/filtration represents a promising alternative to powdered activated carbon and ozone for advanced wastewater treatment.

Straw biochar hastens organic matter degradation and produces nutrient-rich compost.

Biochar derived from wheat straw was added to pig manure in amounts equivalent to 5%, 10%, or 15% (w/w, wet weight). The ratios of NH4(+)/NO3(-) and of UV light absorption at a wavelength of 254nm (SUV254) and dissolved organic carbon (DOC) indicated that compost with 10-15% biochar became more mature and more humified within 42days of composting, and the content of DOC and the concentration of NH4(+) in such compost decreased by 37.5-62.0% and 4.0-20.9%, respectively, compared to the corresponding levels in the control. Addition of biochar lowered the pH and increased electrical conductivity by 7.0-37.5% compared to the control and also increased the concentrations of water-soluble nutrients including PO4(3-) (5.6-7.4%), K(+) (14.2-58.6%), and Ca(2+) (0-12.5%). It is therefore recommended that straw biochar be added to pig manure at 10-15% by weight.

Removal of phosphorus by a high rate membrane adsorption hybrid system.

Membrane adsorption hybrid system (MAHS) was evaluated for the removal of phosphate from a high rate membrane bioreactor (HR-MBR) effluent. The HR-MBR was operated at permeate flux of 30L/m(2)h. The results indicated that the HR-MBR could eliminate 93.1±1.5% of DOC while removing less than 53% phosphate (PO4-P). Due to low phosphate removal by HR-MBR, a post-treatment of strong base anion exchange resin (Dowex(∗)21K-XLT), and zirconium (IV) hydroxide were used as adsorbent in MAHS for further removal of phosphate from HR-MBR effluent. It was found that the MAHS enabled to eliminate more than 85% of PO4-P from HR-MBR effluent. Hence, HR-MBR followed by MAHS lead to simultaneous removal of organics and phosphate in a reliable manner. The experiments were conducted only for a short period to investigate the efficiency of these resins/adsorbents on the removal of phosphorus and high rate MBR for organic removal.

Improving composting as a post-treatment of anaerobic digestate.

This work investigated the influences of practical parameters upon composting of digestate. The yardsticks for evaluation were digestate stabilization, nitrogenous emissions mitigation and self-heating potential. The results suggest choosing an "active" bulking agent like dry wood chips (WC) which served as free-water and nitrogen sink through composting. At an optimal volumetric WC:digestate mixing ratio of 4:1, nearly 90% of the initial NH4(+)/NH3 were fixed, which reduced significantly nitrogenous emissions. This mixing ratio also improved the stabilization and self-heating potential. Using small particle size WC increased narrowly O2 consumption and reduced NH3 emission. Storing used WC prior to recycling reduced 40% N2O emission compared to directly recycled WC. Recycling compost helped to decrease NH3 emission, but quadrupled N2O emission. The optimal aeration rate (15Lh(-1)kg OM0) which was lower compared to composting of organic waste, was enough to ensure the O2 supply and ameliorate the self-heating potential through composting of digestate.

Chemical Constituents of Plants from the Genus Eupatorium (1904-2014).

Eupatorium (family: Compositae), which comprises nearly 1200 species, is distributed throughout tropical America, Europe, Africa, and Asia. Up to now, the reported constituents from the genus Eupatorium involve flavonoids, terpenoids, pyrrolizidine alkaloids, phenylpropanoids, quinonoids, essential oils, and some others, altogether more than 300 compounds. Studies have shown that Eupatorium and its active principles possess a wide range of pharmacological activities, such as cytotoxic, antifungal, insecticidal, antibacterial, anti-inflammatory, and antinociceptive activities. Currently, effective monomeric compounds or active parts have been screened for pharmacological activities from Eupatorium in vivo and in vitro. Increasing amount of data supports application and exploitation for new drug development.

Effluent quality and reuse potential of domestic wastewater treated in a pilot-scale hybrid constructed wetland system.

The study investigates treatment and reuse potential of domestic wastewater of a small community of about 30 people sequentially by anaerobic pretreatment followed by horizontal (HSSF-CW) and vertical (VSSF-CW) sub-surface flow constructed wetlands operated in series. The organic and suspended solids load to the hybrid wetland system was decreased by anaerobic pretreatment. HSSF-CW mainly removed organic matter and supported denitrification whereas VSSF-CW mainly obtained nitrification and phosphorus removal. Recirculation of the effluent increased particularly total nitrogen removal in the wetland system. The study involves evaluation of the whole system in terms of effluent quality. It was achieved on average >95% organic matter and >90% nitrogen removal in the hybrid constructed wetland system with anaerobic pretreatment at a specific wetland surface area of only about 1 m(2) per person. Average mass removal rates were 21.17 gCOD/m(2)day, 5.58 gBOD5/m(2)day, 2.78 gTKN/m(2)day, 1.35 gTN/m(2)day, 0.44 gTP/m(2)day and 5.21 gTSS/m(2)day throughout the total duration of the operation. Consequently, the effluent met the regulations for discharge limits for organic matter and suspended solids. COD and TN concentrations decreased to below 20 mg/L in the effluent. It was also shown that effluent of the system could be reused for irrigation if it is disinfected properly.

Treatment of petroleum refinery wastewater containing heavily polluting substances in an aerobic submerged fixed-bed reactor.

Petroleum refineries produce large amount of wastewaters, which often contain a wide range of different compounds. Some of these constituents may be recalcitrant and therefore difficult to be treated biologically. This study evaluated the capability of an aerobic submerged fixed-bed reactor (ASFBR) containing a corrugated PVC support material for biofilm attachment to treat a complex and high-strength organic wastewater coming from a petroleum refinery. The reactor operation was divided into five experimental runs which lasted more than 250 days. During the reactor operation, the applied volumetric organic load was varied within the range of 0.5-2.4 kgCOD.m(-3).d(-1). Despite the inherent fluctuations on the characteristics of the complex wastewater and the slight decrease in the reactor performance when the influent organic load was increased, the ASFBR showed good stability and allowed to reach chemical oxygen demand, dissolved organic carbon and total suspended solids removals up to 91%, 90% and 92%, respectively. Appreciable ammonium removal was obtained (around 90%). Some challenging aspects of reactor operation such as biofilm quantification and important biofilm constituents (e.g. polysaccharides (PS) and proteins (PT)) were also addressed in this work. Average PS/volatile attached solids (VAS) and PT/VAS ratios were around 6% and 50%, respectively. The support material promoted biofilm attachment without appreciable loss of solids and allowed long-term operation without clogging. Microscopic observations of the microbial community revealed great diversity of higher organisms, such as protozoa and rotifers, suggesting that toxic compounds found in the wastewater were possibly removed in the biofilm.

Nutrients removal and lipids production by Chlorella pyrenoidosa cultivation using anaerobic digested starch wastewater and alcohol wastewater.

The cultivation of microalgae Chlorella pyrenoidosa (C. pyrenoidosa) using anaerobic digested starch wastewater (ADSW) and alcohol wastewater (AW) was evaluated in this study. Different proportions of mixed wastewater (AW/ADSW=0.176:1, 0.053:1, 0.026:1, v/v) and pure ADSW, AW were used for C. pyrenoidosa cultivation. The different proportions between ADSW and AW significantly influenced biomass growth, lipids production and pollutants removal. The best performance was achieved using mixed wastewater (AW/ADSW=0.053:1, v/v), leading to a maximal total biomass of 3.01±0.15 g/L (dry weight), lipids productivity of 127.71±6.31 mg/L/d and pollutants removal of COD=75.78±3.76%, TN=91.64±4.58% and TP=90.74±4.62%.

Influence of a compost layer on the attenuation of 28 selected organic micropollutants under realistic soil aquifer treatment conditions: insights from a large scale column experiment.

Soil aquifer treatment is widely applied to improve the quality of treated wastewater in its reuse as alternative source of water. To gain a deeper understanding of the fate of thereby introduced organic micropollutants, the attenuation of 28 compounds was investigated in column experiments using two large scale column systems in duplicate. The influence of increasing proportions of solid organic matter (0.04% vs. 0.17%) and decreasing redox potentials (denitrification vs. iron reduction) was studied by introducing a layer of compost. Secondary effluent from a wastewater treatment plant was used as water matrix for simulating soil aquifer treatment. For neutral and anionic compounds, sorption generally increases with the compound hydrophobicity and the solid organic matter in the column system. Organic cations showed the highest attenuation. Among them, breakthroughs were only registered for the cationic beta-blockers atenolol and metoprolol. An enhanced degradation in the columns with organic infiltration layer was observed for the majority of the compounds, suggesting an improved degradation for higher levels of biodegradable dissolved organic carbon. Solely the degradation of sulfamethoxazole could clearly be attributed to redox effects (when reaching iron reducing conditions). The study provides valuable insights into the attenuation potential for a wide spectrum of organic micropollutants under realistic soil aquifer treatment conditions. Furthermore, the introduction of the compost layer generally showed positive effects on the removal of compounds preferentially degraded under reducing conditions and also increases the residence times in the soil aquifer treatment system via sorption.

Performance of nitrate-dependent anaerobic ferrous oxidizing (NAFO) process: a novel prospective technology for autotrophic denitrification.

Nitrate-dependent anaerobic ferrous oxidizing (NAFO) is a valuable biological process, which utilizes ferrous iron to convert nitrate into nitrogen gas, removing nitrogen from wastewater. In this work, the performance of NAFO process was investigated as a nitrate removal technology. The results showed that NAFO system was feasible for autotrophic denitrification. The volumetric loading rate (VLR) and volumetric removal rate (VRR) under steady state were 0.159±0.01 kg-N/(m(3) d) and 0.073±0.01 kg-N/(m(3) d), respectively. In NAFO system, the effluent pH was suggested as an indicator which demonstrated a good correlation with nitrogen removal. The nitrate concentration was preferred to be less than 130 mg-N/L. Organic matters had little influence on NAFO performance. Abundant iron compounds were revealed to accumulate in NAFO sludge with peak value of 51.73% (wt), and they could be recycled for phosphorus removal, with capacity of 16.57 mg-P/g VS and removal rate of 94.77±2.97%, respectively.