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Experimental Design for the Effect of Inorganic Compound in the Aerobic Granular Sludge Development
Master Student: Muhammad Anwar Alias Supervisor: Dr. Khalida Bt Muda Co- Supervisor: Dr. Azmi Aris
Outline of Presentation
Introduction
Problem Statement
Aim and Objectives of the Study Scope of the Study Significance of the Research Literature Review Methodology Expected Outcomes
Introduction
Biorranulation
Granulation is the process in which a non discrete flocculent biomass forms discrete welldefined pellets or granules which can be several millimeters in diameter (Ong et al., 2002). Involves cell-to-cell interactions that include biological, physical and chemical phenomen
Introduction
Biogranulation can be classified as: aerobic granulation anaerobic granulation Biogranules form through self-immobilization of microorganisms. These granules are dense microbial consortia packed with different bacterial species and typically contain millions of organisms per gram of biomass.
Problem Statement
Time consuming for developing fully complete matured granules.
During long-term SBR operation, the aerobic granular sludge process gradually loses stability under inappropriate operational conditions. Aerobic granules often showed a poor stability after several months of operation (de Kreuk and van Loosdrecht, 2004; Liu et al., 2006) which the application wastewater treatment is limited
1
To identify the potential soluble ionic inorganic compounds that contributes in accelerating biogranulation formation during start up period
2
To optimize the effect of inorganic compound as aid in the biogranulation development
3
To develop biogranules with the additional of inorganic compound
Scope
This research could be helpful for enhancing the aerobic granules development via addition of ionic organic compound as granulation aid in terms of fast formation, high durability and stability, and high removal performance. Thus, this research development can be utilized to overcome our local waste water problem especially in palm oil mill effluent, leachate and municipal waste water.
Literature Review
Aerobic Sludge Granulation Process
Sludge may be defined as the microbial biomass utilizing nutrient substrates present in wastewater. Microbial granules are characterized as dense and compact microbial aggregates with a spherical outer shape. Aerobic granules are self-immobilized spherical aggregates of microorganisms. The growth of aerobic granules were considered to be a special case of biofilm growth (Liu and Tay, 2002; Yang et al., 2004).
Literature Review
Inorganic Compound
Inorganic compounds have traditionally been considered to be of inanimate, non-biological origin Inorganic compounds, consist of molecules with a central metal atom (usually transition metal) bonded to one or more non-metallic ligand(inorganic, organic, or both) and are often intensely coloured In the form of ion, its acts as electron-transfer proteins and substrate bindings Example : MgSO4, Al(NO3)39H2O, BaCl2, Pb(NO3)2, MnSO4H2O
Literature Review
Extracellular Polymeric Substance (EPS) Also known as exopolysaccharides EPS are a complex high-molecular-weight mixture of polymers excreted by microorganisms/microbes Produced from cell lysis and adsorbed organic matter from wastewater
Literature Review
EPS is recognized as a key factor in the granulation process (Wang et al. 2006), which is mainly composed of polysaccharide and protein Previous studies shows divalent metal ionic such as Ca2+, Mg2+ and Fe2+ augmentation led to the higher production yield of EPS (polysaccharides & protein) resulted in faster granulation process and shorter starting-up period
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Literature Review
Previous studies shows that Cationic from the inorganic compound :
can neutralize the negative charge on the surface of bacteria, and promote the sludge granulation because of the positive charge The cationic could bind to the negatively charged groups on bacterial surfaces and extracellular polymeric substances (EPS), and act as a bridge to interconnect these components, and then form EPS(cationic)EPS crosslinkage could accelerate microbial aggregation, and increase the physical strength of mature granules
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Based on experiments in an aerobic upflow sludge blanket (AUSB), the biological assemblies is similar to the anaerobic granulation process, filamentous bacteria tangled with each other to form aerobic granules.
Negatively Charged
- Individual bacterium
+ + +
BRIDGING
SHAPING
+ - +
LITERATURE REVIEW
Previous studies on sludge biogranulation process using inorganic compound
Performance on granulation development 25 days Reactor system Biomass Type of wastewater Synthetic waste water Inorganic compound used R1 augmented with 10 mg/L of Mn2+ R2 no Mn2+ Biogranule characteristic Diameter size: R1= 2.53.5 mm R2= 1.5- 2.5mm Remarks Aerobic granules first formed in reactor R1 on day 6 Ca2+-fed granules were denser and more compact, better settling and strength characteristics, and higher polysaccharide contents. The researches had indicated that an increase in granule size resulted in the Ca2+ accumulation in granules, and finally led to a reduced granule bioactivity [ Reference
SBR
Aerobic granule
Lihui et al (2011)
ASB
Aerobic granules
Synthetic wastewater
Ca2+
augmentation
32 days
SBR
Aerobic granule
21 days
UASB = Upflow Anaerobic Sludge Blanket; SBR = Sequencing Batch Reactor; ASB = Sequential Aerobic Sludge Blanket; SS = Suspended Solid; VSS = Volatile Suspended Solid; MLSS = Mixed Liquor Suspended Solid; MLVSS = Mixed Liquor Volatile Suspended Solid; SV = Settling Velocity; SVI = Sludge Volume Index; SAA = Specific Acidogenic Activity; SMA = Specific Methanogenic Activity; EPS = Extracellular Polymeric Substance; CONTINUED.. COD = Chemical Oxygen Demand; TOC = Total Organic Content; NH4+-N = Ammonium
CONTINUED..
LITERATURE REVIEW
Type of wastewater Synthetic wastewater (glucose) Influent COD = 640 mg/l Synthetic wastewater (potassium nitrate; methanol) Nitrate = 120 mg/l; TOC = 120 mg/l Biomaterial used Biogranule characteristic Size: 2.9mm Specific gravity: 1.002 kg/l MLSS: 3.2 g/l SS: 7.6 g/l SVI: 25 ml/g Performance removal efficiency Effluent COD = 56 mg/l; COD removal = 91% Remarks Mg2+ augmentation significantly decreased the sludge granulation time from 32 days to 18 days. Reference
SBR
Mg2+ augmentation
SBR
Mean size of biomass: 420m 2+ augmentation MLVSS/MLSS Ca ratio: 0.37 SVI: 14.36 ml g-1
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Literature Review
Previous studies shows that Cationic from the inorganic compound :
can neutralize the negative charge on the surface of bacteria, and promote the sludge granulation because of the positive charge The cationic could bind to the negatively charged groups on bacterial surfaces and extracellular polymeric substances (EPS), and act as a bridge to interconnect these components, and then form EPS(cationic)EPS crosslinkage could accelerate microbial aggregation, and increase the physical strength of mature granules
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Methodology
Single colonies of isolated bacteria will be labeled and stored in universal bottle as stocks.
The stocks are stored at 4C and working solutions of required concentrations were obtained by appropriate dilution in deionized water prior to each experiment Culture will be grown in the Nutrient Broth to more quantity
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Research Framework
Objective 1
Stage 1 To identify the potential of soluble ionic inorganic compound as aid for the aerobic granulation start-up
Isolation of bacteria from sewage waste water sludge
Screening for bacteria selection in the aerated synthetic medium Response: Hydrophobicity and Autoaggregation
Stage 2
Screening for soluble inorganic compound using aerated synthetic medium and selected bacteria in room temperature Variables: Type of ionic inorganic compound (based on concentration) Response: Hydrophobicity and Auto aggregation
Stage 3
Data analysis using Completely Randomized Design (CRD) method to select the best ionic inorganic compound
Research Framework
Objective 2 To optimize the effect of inorganic compound as aid in
granulation start up using response surface method Optimize the selected ionic inorganic compound using synthetic medium with aeration under room temperature Variables: Concentration, pH and temperature Response: Extracellular polymer substance(EPS) secretion, Hydrophobicity and Auto aggregation Data analysis using Response Surface method to optimize the best condition of ionic inorganic compound for the aerobic granulation start-up
Research Framework
Objective 3
To develop biogranules with the additional of soluble inorganic compound
Operation process using according to Muda K. et al. 2009 method + (inorganic compound) i. Granular sludge properties ii. Granular sludge removal performance
Experiment Set up
Objective 1 500mL of Schott bottle will be used as reactor with a working volume of 250 mL (synthetic medium + bubbling for aeration + culture in nutrient broth,10%(v/v)) + (inorganic compound) Batch process - (R1, R2, R3, R4, R5, R6 and R7) respectively will be augmented with Al(NO3)39H2O, BaCl2, Pb(NO3)2, MnSO4H2O, ZnSO47H2O, NiSO6H20 and Sr(NO3)29H2O Medium Synthetic Waste water (according to khalida muda et al. 2011 method) Air flow rate for aeration = 1Lmin-1 pH = 6.0 8.0 Temperature = 272 0C (room temperature) Duration = 30min
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Experiment Set up
Objective 2 500mL of Schott bottle will be used as reactor with a working volume of 250 mL (synthetic medium + bubbling for aeration + culture in nutrient broth,10%(v/v)) + (inorganic compound) Medium Synthetic Waste water (according to khalida muda et al. 2011 method)
Duration = 60 min
Air flow rate for aeration = 1Lmin-1 Variable parameters:
- Temperature = 27- 30 0C
- pH= 6-8 - Concentration of inorganic compound selected from the 1st objective
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Experiment Set up
Objective 3
Parameter Measurements
Objective 1
Parameter Measurements
Objective 2
Autoaggregation Assay - Turbidity Meter- refer to khalida muda et al. 2011 Surface Hydrophobicity Assay - Optical Density Meter- refer to khalida muda et al. 2011 Characteristics of the Extracellular Polymeric Substance (EPS) and protein method will refer to Zhang et al. 1999 & Liu and Fang, 2002 Alginate-like exopolysaccharides (ALE) extraction and identification method will refer to Lin et al.2008
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Expected Outcomes
Expected Results
The potential soluble inorganic compounds that contributes in accelerating biogranulation formation during start up period were identified
The optimum condition for the effect of soluble inorganic compound as aid in the biogranulation development were achieved
Aerobic granules were developed with the additional of soluble inorganic compound
Expected Outcomes
Negatively Charged
Individual bacterium
- + BRIDGING
+
+
+ + +
SHAPING
+ - +
any anion
nitrate NO3acetate (CH3COO-) Chloride (Cl-), Bromide (Br-), Iodide (I-) Sulphate (SO42-)
+
+ + + + +
=
= = = =
soluble
soluble soluble low solubility (insoluble) soluble low solubility (insoluble)
calcium (Ca2+), strontium (Sr2+), barium (Ba2+), silver = (Ag+), lead (Pb2+), radium (Ra2+) any other cation alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+), alkali earth metals (Be2+,Mg2+,Ca2+,Sr2+,Ba2+,Ra2+), and H+(aq), NH4+ any other cation alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+), H+(aq),NH4+,Sr2+,Ba2+,Ra2+,Tl+ any other cation alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+), H+(aq),NH4+ = =
+ Sulfide S2+
soluble soluble
Hydroxide OH-
+ + +
= = = =
zinc sulfide, ZnS, is insoluble strontium hydroxide, Sr(OH)2, is soluble silver hydroxide, AgOH, is insoluble (forms a precipitate) ammonium phosphate, (NH4)3PO4, is soluble magnesium carbonate, MgCO3, is insoluble