Granulation, P Granulation, P - - Dynamics, and Dynamics, and Population Shifts in a Column Population Shifts in a Column SBR operated for EBPR SBR operated for EBPR Ebru Dulekgurgen Ebru Dulekgurgen 08.06.04 (UIUC) 08.06.04 (UIUC) Background pictures: Unpublished data by E. Dulekgurgen (2004) Background pictures: Unpublished data by E. Dulekgurgen (2004)
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Granulation, PGranulation, P--Dynamics, and Dynamics, and Population Shifts in a Column Population Shifts in a Column
SBR operated for EBPR SBR operated for EBPR
Ebru DulekgurgenEbru Dulekgurgen08.06.04 (UIUC)08.06.04 (UIUC)Background pictures: Unpublished data by E. Dulekgurgen (2004)Background pictures: Unpublished data by E. Dulekgurgen (2004)
Column SBRColumn SBR
ColumnColumn--type SBR, H/D=74.4/7.2cmtype SBR, H/D=74.4/7.2cmVwVw = 3L = 3L ExchangeExchange =50%=50%QairQair = 150 L/hr= 150 L/hrPowerPoweraeraer =0.26 Watt =0.26 Watt GGaeraer = 208 sec= 208 sec--11
Influent: Influent: 400 mgCOD/L (acetate)400 mgCOD/L (acetate)17 mgPO17 mgPO44
--33--P/LP/LOperation:Operation:
2hr Anaerobic/ 3hr 44min Aerobic/12hr Anaerobic/ 3hr 44min Aerobic/1--5min settling5min settling
Column SBRColumn SBRInoculum (D01): Inoculum (D01):
EBPR biomass from CarlosEBPR biomass from Carlos’’s SBRs SBR
Granular Biomass (D99)Granular Biomass (D99)
Augmentation (D40Augmentation (D40--D47):D47):EBPR biomass from CarlosEBPR biomass from Carlos’’s SBRs SBR
Granulation strategy: Granulation strategy: High shear, low settling time (ts), High shear, low settling time (ts), slowslow--growers (PAOs)growers (PAOs)
Biomass/SettlingBiomass/Settling
ColSBR: SVI vs MLSS
0
500
1000
1500
2000
2500
3000
3500
4000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Time (days)
MLS
S (m
g/L)
0
50
100
150
200
250
300
350
SVI (
mL/
g)
MLSS (mg/L) SVI (mL/g)
ts5min
ts:1 min ts: 5min ts:1 min
ColSBR: Effluent SS
0
20
40
60
80
100
120
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Time (days)
Con
c. (m
g/L)
ts5min
ts:1 min ts: 5min ts:1 min
EffluentEffluent
CC--DynamicsDynamics
ColumnSBR: Carbon Removal
0
100
200
300
400
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Time (days)
Solu
ble
CO
D (m
g/L)
Effluent Anaerobic
Start-up
Augmentation1+2: Foaming and biofilm formation stoppped
ts5min
T:26oC 29oC 22oC 22oC 21oC
ts:1 min
Biomass washoutExtensive FOAMINGExtensive BIOFILM1st granulation
ts: 5min ts:1 min
After Augmentation:2nd granulationCycle profilesP-dynamics
POPO44--33--P DynamicsP Dynamics
ColumnSBR: PO4-3-P Removal
0
10
20
30
40
50
60
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Time (days)
PO4-3
-P (m
g/L)
Effluent Anaerobic
Start-up
Augmentation1+2: Foaming and biofilm formation stoppped
ts5min
T:26oC 29oC 22oC 22oC 21oC
ts:1 min
Biomass washoutExtensive FOAMINGExtensive BIOFILM1st granulation
ts: 5min ts:1 min
After Augmentation:2nd granulationCycle profilesP-dynamics
Column SBR: Extracellular Polysaccharides (ECP) and Extracellular Proteins (E Prot)
0
100
200
300
400
500
600
700
D01 D08 D39 C39 D56 D100Time (days)
Con
c. (m
g/L)
EPSac (mg glucose/L) EProt (mg BSA/L)
Extracellular polysaccharides and proteins per unit biomass
0.0
0.1
0.2
0.3
0.4
0.5
0.6
D01 D08 D39 C39 D56 D100Time (days)
Valu
es n
orm
aliz
ed to
MLV
SS
EPSac (mg glucose/mg MLVSS) EProt (mg BSA/mg MLVSS)
EPS EPS Extracellular polysaccharides and ProteinsExtracellular polysaccharides and Proteins
D01D01 : Inoculum: InoculumD08D08 : Before ts decreased to 1 min: Before ts decreased to 1 minD39D39 : Before augmentation: Before augmentationC39C39 : Biomass for augmentation (Carlos: Biomass for augmentation (Carlos’’s SBR)s SBR)D56D56 : Before ts decreased to 1 min: Before ts decreased to 1 minD100D100 : Granular biomass: Granular biomass
EPS EPS Extracellular polysaccharides and ProteinsExtracellular polysaccharides and Proteins
x8x8 x8x8 x10x10 x6x6
EPSac / EProt (mg glucose/mg BSA)57.0% 55.8%
39.5%41.9%
44.5%
38.7%
0%
10%
20%
30%
40%
50%
60%
D01 D08 D39 C39 D56 D100Time (days)
ECP/
E Pr
ot
TRFLP to Monitor the PopulationsTRFLP to Monitor the Populations
TRFLP? TRFLP? Terminal Restriction Fragment Length PolimorphismTerminal Restriction Fragment Length Polimorphism
Extract genomic DNAExtract genomic DNAAmplify region on SSU rDNA (PCR)Amplify region on SSU rDNA (PCR)Use FluoUse Fluo--labeled primer (PCR: *Eub338 and Univ1392a)labeled primer (PCR: *Eub338 and Univ1392a)Digest amplicons w/ restriction enzymesDigest amplicons w/ restriction enzymesElectrophore the digets Electrophore the digets
TRFLP to Monitor the PopulationsTRFLP to Monitor the Populations
ElectophorogramsElectophorograms
GoodGood
LowLow
Monomorphic!!Monomorphic!!
Polymorphic but;Polymorphic but;
TRFLP to Monitor the PopulationsTRFLP to Monitor the Populations
In silico TRFLP: Results of amplification and DigestionIn silico TRFLP: Results of amplification and Digestion
HaeIII-diversity
0
100
200
300
400
500
600
700
800
900
1000
0 250 500 750 1000 1250 1500 1750 2000 2250 2500 2750 3000 3250 3500 3750 4000 4250 4500 4750 5000 5250 5500 5750 6000 6250 6500 6750 7000 7250 7500 7750 8000 8250 8500 8750 9000
Seq. Order
Frag
men
t Len
gth
TRFLP to Monitor the PopulationsTRFLP to Monitor the Populations
Electrophorogram for PAO population Electrophorogram for PAO population in the Column SBR (Day1in the Column SBR (Day1--18)18)
D1D1
D4D4
D8D8
D13D13
D18D18
Preliminary TRFLP ResultsPreliminary TRFLP Results
Significant Results for Significant Results for PAOs PAOs andand GAOsGAOs
PAOsPAOs
GAOsGAOs
130
1.37% 2.45%0 0
2.52%
15.99%
40.48%40.20%42.90%
4.05%
23.07%
16.21%
0 0 1.20% 0 0 1.51%0%5%
10%15%20%25%30%35%40%45%50%
D01 D04 D08 D13 D18 D20 D29 D34 D40 C40 D43 D45 D47 D50 D62 D70 D84 D98
Time (days)
% P
eak
Are
a
410+415
53.40%
35.81%41.93%
38.35%
1.33% 0.00% 0.00% 0.00% 0.00%
29.18%
18.56%
30.32%
45.83%
24.45%
43.24%
62.54%
54.75%
21.20%
0%
10%
20%
30%
40%
50%
60%
70%
D01 D04 D08 D13 D18 D20 D29 D34 D40 C40 D43 D45 D47 D50 D62 D70 D84 D98
Time (days)
% P
eak
Are
a
AcknowledgementsAcknowledgements
Eberhard Eberhard MORGENROTH, MORGENROTH, Lut Lut RASKIN, RASKIN, Julie Julie ZILLESZILLESRocky BARIONES, Dominic FRIGON, Adam Rocky BARIONES, Dominic FRIGON, Adam KLEIN, Carlos LOPEZ, Sudini PADMASIRI, Rob KLEIN, Carlos LOPEZ, Sudini PADMASIRI, Rob SANFORD, Chuanwu XISANFORD, Chuanwu XI..........and all the members of the ..........and all the members of the
MORGENROTH Group and RASKIN Group MORGENROTH Group and RASKIN Group
Nazik ARTAN, Derin ORHONNazik ARTAN, Derin ORHON
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