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SUPPORTING INFORMATIONEnhanced aerobic sludge granulation by applying carbon fibers as nucleating skeletons Jie Xua, *, Heliang Pang a, Junguo Heb,* * , Mengfei Wanga, Jun Nan a , Lin Liaa School of Environment, Harbin Institute of Technology (HIT), Harbin 150090, Chinab School of Civil Engineering, Guangzhou University, Guangzhou 510006, China*Corresponding author, Tel: +86-0451-86289099. E-mail address: xujiehit@ (Jie Xu)Address: Second campus of HIT, No.73 Huanghe Road, Nangang District, Harbin, China.*Corresponding author, Tel: +86-020-39366166. E-mail address: hejunguohit@ (Junguo He)Address: 230 West Ring Road, Guangzhou University Town, Panyu District, Guangzhou, China.Extraction of EPSA sludge suspension was first dewatered by centrifugation (5810R, Eppendorf) in a 50-mL tube at 4000g for 5 min. The sludge in the tube was then resuspended into 15 mL of 0.05% NaCl solution that had a similar salinity to the solution in the AGS reactor. The sludge mixture was then diluted with the NaCl solution to its original volume of 50 mL. The NaCl solution for dilution was pre-heated to 70 °C to ensure that the sludge suspension reached an immediate warm temperature of 50 °C. Without any delay, the sludge suspension was then sheared by a vortex mixer (Maxi Mix II, Thermolyne) for 1 min, followed by centrifugation at 4000g for 10 min. The organic matter in the supernatant was readily extractable EPS, and was regarded as the LB-EPS of the biomass.For the extraction of the TB-EPS, the sludge left in the centrifuge tube was resuspended in 0.05% NaCl solution to its original volume of 50 mL. The sludge suspension was heated to 60 °C in a water bath for 30 min, and the sludge mixture was then centrifuged at 4000g for 15 min. The supernatant that was collected was regarded as the TB-EPS extraction of the sludge.Staining and confocal laser scanning microscopy (CLSM) imagingIn staining, Syto 63 (20 μM, 100 μl) was first added to the sample that was placed on a shaker table for 30 min. Next, 0.1-M sodium bicarbonate buffer (100 μl) was added to maintain the amine group in non-protonated form that was followed by a FITC solution (10 g/l, 10 μl), and the mixture stirred at room temperature (1 h). Subsequently, the calcofluor white (fluorescent brightener 28, 300 mg/l, 100 μl) was incubated with the sample for another 30 min to stain the β-linked D-glucopyranose polysaccharides. Table S1 Characteristics of AGS at different operating period. Operating dayMLSS (g/L)MLVSS/MLSS (%)SVI30 (mL/g)Granulation phase (days)R1R2R3R1R2R3R1R2R304.54.54.5515151112112112Ⅰ (0-30)Ⅱ (30-40)Ⅲ (40-90)Ⅳ (90-120)Ⅴ (120-150)301.61.83.0817967626963401.72.52.86177721075245901.55.05.567767315040301204.56.06.57573716045401505.06.26.4777572563531Operating dayCOD (%)TN (%)TP (%)R1R2R3R1R2R3R1R2R3091919150505010101030899697525351132514408596983858501727269090949851606069999120939594776460979799150949593736154999991Table S2 Bacterial community richness and diversity indices for different sludge samples.Sample IDOTU numberChao indexACEindexShannon indexSimpsonSEED1032109510755.730.007R14485235234.140.035R1-24565515484.010.041R26507557594.410.048R2-26057386993.510.095R3R3-25685496456856336564.473.810.0310.069utsrqponmlkjihgfedcbaFigure S1 Images of granules at different stage of the granulation process: seed sludge (a); short CFs (b); long CFs (c); day 30 (d-f); day 40 (g-i); day 90 (j-l); day 120 (m-o); SEM at low magnification (p-r) and high magnification (s-u).abcFig S2 Effluent concentrations of ammonia, nitrite and nitrate in R1 (a), R2 (b) and R3 (c).Figure S3 Pollutants removal processes in typical cycles: day 80 (a, c, e) and day 140 (b, d, f).Figure S4 3D-EEM of LB-EPS and TB-EPS in seed sludge (a, b), R1 (c, d), R2 (e, f) and R3 (g, h). ................
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