Anwar Slide Presentation

8/2/2019 Anwar Slide Presentation

1/38

Experimental Design for the Effect of InorganicCompound in the Aerobic Granular SludgeDevelopment

Research Proposal: First assessment

Master Student: Muhammad Anwar Alias

Supervisor: Dr. Khalida Bt Muda

Co- Supervisor: Dr. Azmi Aris


2/38

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


3/38

Introduction

Biorranulation

Granulation is the process in which a nondiscrete flocculent biomass forms discrete well-

defined pellets or granules which can be severalmillimeters in diameter (Ong et al., 2002).

Involves cell-to-cell interactions that includebiological, physical and chemical phenomen

Granulation aid is promoting the formation of thegranules or granular biomass.


4/38

Introduction

Biogranulation can be classified as: aerobic granulation anaerobic granulation

Biogranules form through self-immobilization ofmicroorganisms.

These granules are dense microbial consortiapacked with different bacterial species andtypically contain millions of organisms per gramof biomass.


5/38

Advantages of aerobic granules More compact microbial structure Improved settling ability.

Enable a high biomass retention. Withstand high-strength wastewater & shock

loadings. Effective sludge-effluent separation Greater biomass preservation and enrichment

significantly higher organic loading rates

Factors affect aerobic granulation Organic loading rate Seed sludge

Aeration intensity Settling time Granulating agent Hydraulic retention time

Divalent metal ions


6/38

Problem Statement

Time consuming for developingfully complete matured granules.

During long-term SBR operation, the aerobic

granular sludge process gradually loses stabilityunder inappropriate operational conditions.

Aerobic granules often showed a poor stabilityafterseveral months of operation (de Kreuk andvan Loosdrecht, 2004; Liu et al., 2006) which theapplication wastewater treatment is limited


7/38

Aim and Objectives of the Study

1To identify thepotential solubleionic inorganiccompounds thatcontributes in

acceleratingbiogranulationformation duringstart up period

2

To optimize theeffect of inorganiccompound as aidin thebiogranulation

development

3To developbiogranules withthe additional ofinorganiccompound

The aim of this study is to enhance the aerobic granulation processusing inorganic compound.


8/38

Scope of the Study

Scope

Start up for aerobic granulationprocess/Initial stage

InorganicCompound as aid

Lab scale reactoraerobic condition

Bacterial isolation fromsewage sludgeSynthetic wastewater

Parameters to studythe affect

Apply to the existencesystem- develop

granule


9/38

Significance of the Research

This research could be helpful for enhancing the aerobicgranules development via addition ofionic organic compound

as granulation aid in terms of fast formation, high durabilityand stability, and high removal performance.

Thus, this research development can be utilized to overcome ourlocal waste water problem especially in palm oil mill effluent,leachate and municipal waste water.


10/38

Literature Review

Aerobic Sludge Granulation Process

Sludge may be defined as the microbial biomassutilizing nutrient substrates present in wastewater.

Microbial granules are characterized as dense

and compact microbial aggregates with aspherical outer shape.

Aerobic granules are self-immobilized sphericalaggregates of microorganisms.

The growth of aerobic granules were considered

to be a special case of biofilm growth (Liu andTay, 2002; Yang et al., 2004).


11/38

Inorganic Compound Inorganic compounds have traditionally been

considered to be of inanimate, non-biological origin

Inorganic compounds, consist of molecules with acentral metal atom (usually transition metal) bondedto 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 proteinsand substrate bindings

Example : MgSO4, Al(NO3)39H2O, BaCl2, Pb(NO3)2,MnSO4H2O

Literature Review


12/38

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


13/38

Literature Review

EPS is recognized as a key factor in thegranulation process (Wang et al. 2006),which is mainly composed ofpolysaccharide and protein

Previous studies shows divalent metalionic such as Ca2+, Mg2+ and Fe2+

augmentation led to the higher productionyield ofEPS (polysaccharides & protein)resulted in faster granulation process andshorter starting-up period


14/38

can neutralize the negative charge on the surface ofbacteria, and promote the sludge granulation because ofthe positive charge

The cationic could bind to the negatively charged groups onbacterial surfaces and extracellular polymeric substances(EPS), and act as a bridge to interconnect thesecomponents, and then form EPS(cationic)EPS cross-linkage

could accelerate microbial aggregation, and increase thephysical strength of mature granules

www.themegallery.com

Company Logo

Literature Review

Previous studies shows that Cationic from the inorganic

compound :


15/38

Mechanism of aerobic granulation


16/38

Mishimaand

Nakamura(1991)

Based on experiments in an aerobic upflow sludge blanket(AUSB), the biological assemblies is similar to the anaerobicgranulation process, filamentous bacteria tangled with eachother to form aerobic granules.

Beun et al.(1999)

Proposed mechanism of aerobic granulation in SBR

Beun et al. (1999) proposed a model in which aerobicgranulation could be started with fungi.


17/38

Modified schematic representation of extracellular polymeric substance-enhanced

(Liu et al., 2004).

Individual

bacterium

Extracellular polymeric

substance

Positively Charged (Ionic

compound)

Negatively Charged

--

-

- -

-

-

-

-

-

-

-

-+

+

+

+

+

++

BRIDGING

-

-

SHAPING

--

-

-

-

-

-

-

-+

+

+

+

+

+

+

++

+

LITERATURE REVIEW


18/38

Previous studies on sludge biogranulation process using inorganic compoundLITERATURE REVIEW

CONTINUED..

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;COD = Chemical Oxygen Demand; TOC = Total Organic Content; NH4

+-N = Ammonium

Reactor

system

BiomassType of

wastewater

Inorganic

compound used

Biogranule

characteristic

Performance

on granulation

development

Remarks Reference

SBRAerobic

granule

Synthetic

waste water

R1 augmented with

10 mg/L of Mn2+

R2 no Mn2+

Diameter size:

R1= 2.53.5 mm

R2= 1.5- 2.5mm

25 days

Aerobic granules first

formed in

reactor R1 on day 6

Lihui et al

(2011)

ASBAerobic

granules

Synthetic

wastewaterCa2+ augmentation

Diameter: 2.8mm

Wet density: 1.01 kgL-1 32 days

Ca2+-fed granules were

denser and more

compact, better

settling and strength

characteristics, and

higher polysaccharide

contents.

Tay T-L et

al., (2002)

SBRAerobic

granule

Synthetic

waste water

R1 augmented with

Ca2+ at 40 mg/L

R2 augmented withMg2+ at 40 mg/L

Diameter size :

R1= 1.32mm

R2=0.2

1mm

21 days

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[

Lin Liu et

al. (2010)

CONTINUED LITERATURE REVIEW


19/38

Reactorsystem

BiomassType of

wastewaterBiomaterial used

Biogranulecharacteristic

Performanceremoval

efficiencyRemarks Reference

SBRAerobicsludgegranule

Syntheticwastewater(glucose)Influent COD= 640 mg/l

Mg2+augmentation

Size: 2.9mmSpecific gravity:1.002 kg/lMLSS: 3.2 g/lSS: 7.6 g/lSVI: 25 ml/g

Effluent COD= 56 mg/l;COD removal= 91%

Mg2+ augmentationsignificantlydecreased thesludge granulationtime from 32 daysto 18 days.

Li X-M etal., (2008)

SBR

Denitrifyinggranules(anaerobiccondition)

Syntheticwastewater

(potassiumnitrate;methanol)Nitrate = 120mg/l;TOC = 120mg/l

Ca2+ augmentation

Mean size ofbiomass: 420mMLVSS/MLSSratio: 0.37SVI: 14.36 ml g-1

NO3--N; TOC

removal =nearly 100%

-Liu Y-J and

Sun DD,(2010)

CONTINUED.. LITERATURE REVIEWPrevious studies on sludge biogranulation process using inorganic compound


20/38

Literature Review

can neutralize the negative charge on the surface ofbacteria, and promote the sludge granulation because ofthe positive charge

The cationic could bind to the negatively charged groups onbacterial surfaces and extracellular polymeric substances(EPS), and act as a bridge to interconnect thesecomponents, and then form EPS(cationic)EPS cross-linkage

could accelerate microbial aggregation, and increase thephysical strength of mature granules


Company Logo

Previous studies shows that Cationic from the inorganic

compound :


21/38

Methodology


22/38

Stock solutions having 7 types of inorganiccompounds are prepared by dissolvingappropriate quantities in deionized water

List of inorganic compound will be used in this

research:

Preparation of Inorganic compound

1. Aluminium Nitrate Anhydrous

2. Barium Chloride- dihydrate

3. Lead Nitrate

4. Manganese Sulphate-1-hydrate

5. Zinc Sulphate-7-hydrate

6. Nickel Sulphate

7. Strontium Nitrate


23/38

Culture of bacteria is isolated from sewage waste water sludge

Bacteria is isolated on a spread plate.

Single colonies of isolated bacteria will be labeled and stored inuniversal bottle as stocks.

The stocks are stored at 4C and working solutions of requiredconcentrations were obtained by appropriate dilution in deionizedwater prior to each experiment

Culture will be grown in the Nutrient Broth to more quantity

Preparation for stock culture of bacteria


24/38

List of reagent used in the experiment


Company Logo

Reagents/ Chemicals Application

Nutrient Agar Spread Plate

Potassium phosphate, K2HPO4 Washing buffer

Xylene (C6H4(CH3)2) Surface hydrophobicity assay

Nutrient broth Bacteria culture

Sodium Chloride EPS Extraction Method


25/38

Research Framework

Objective 1

Stage 2

To identify the potential of soluble ionic inorganic

compound as aid for the aerobic granulation start-up

Stage 1 Isolation of bacteria from sewage waste water sludge

Screening for bacteria selection in the aerated synthetic medium

Response: Hydrophobicity and Autoaggregation

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 toselect the best ionic inorganic compound


26/38

To optimize the effect of inorganic compound as aid in

granulation start up using response surface method

Objective 2

Optimize the selected ionic inorganic compound using

synthetic medium with aeration under room temperature

Research Framework

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 conditionof ionic inorganic compound for the aerobic granulation start-up


27/38

Research Framework

To develop biogranules with the additional of solubleinorganic compound

Objective 3

Operation process using according to Muda K. et al.

2009 method + (inorganic compound)

i. Granular sludge properties

ii. Granular sludge removal performance


28/38

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


29/38


Company Logo

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


30/38

Operation set up will be according toKhalida Muda et al. 2011 et al


Experiment Set up

Objective 3


31/38

Parameter Measurements

Autoaggregation Assay - Turbidity Meter

Surface Hydrophobicity Assay - Optical Density

Meter

Objective 1


32/38

Parameter Measurements

Autoaggregation Assay - Turbidity Meter- refer to khalida

muda et al. 2011

Surface Hydrophobicity Assay - Optical Density Meter- referto 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

Objective 2


33/38

Experimental Design for data analysis

Objective 1:For screening purpose

Completely Randomized Design ( StatisticalPackage for the Social Science- SPSSsoftware)

Ojective 2: For optimization

Response Surface Method (Design Expert)


Company Logo

Gantt Chart of Research Activities


34/38

Gantt Chart of Research Activities2011 2012 2013

NO ACTIVITY 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 51

Literature Review2 To identify the potential of soluble inorganic

compound as aid for the aerobic granular

sludge developmenta. isolate mixed culture bacteria and 7 identifiedinorganic compound: -

b. Experimental work using CompletelyRandomized Design Method

i. Factors: Type of inorganic compounds

ii. Sample preparation: - the isolated bacteriagrown in schott bottle with volume 250ml ofsynthetic medium + inorganic compound (10% v/v)

iii. Parameter/ Response: coaggregation,hydrophobicity

3 To optimize the effect of inorganic compoundas aid in granulation development usingresponse surface methoda. Experimental work using Response SurfaceMethod

ii. Sample preparation: - the isolated bacteriagrown in schott bottle with volume 250ml ofsynthetic medium +inorganiccompound+aeration(bubbling)

iv. Factors: Concentration of selected inorganiccompounds, temperature and pH

iii. Parameter: EPS, ALE, hydrophobicity andcoaggregation & others

4 to develop biogranules with additional ofinorganic compound (biogranulation aidsolution)a. using Muda K. et al. 2010 method + (inorganiccompound)

i. Granular sludge properties

ii. Granular sludge removal performance

5Thesis Writing


35/38

Expected Outcomes

ExpectedResults

The potential solubleinorganic compoundsthat contributes inacceleratingbiogranulation

formation during startup period wereidentified

Aerobic granuleswere developed withthe additional ofsoluble inorganiccompound

The optimum

condition for theeffect of solubleinorganic compoundas aid in thebiogranulationdevelopment wereachieved


36/38

Modified schematic representation of extracellular polymeric substance-enhanced(Liu et al., 2004).

Individual

bacterium

Extracellular polymeric

substance

Positively Charged (Ionic

compound)

Negatively Charged

--

-

--

-

-

-

-

-

-

-

-+

+

+

+

+

+

+

BRIDGING

-

-SHAPING

--

-

-

-

--

-

-+

+

+

+

+

+

+

++

+

Expected Outcomes


37/38

The Solubility of Inorganic Compounds in Water


38/38

y g p

Negative Ions

(Anions)

+ Positive Ions

(Cations)

= Solubility of compounds in

water

Example

any anion + alkali ions

(Li+,Na+,K+,Rb+,Cs+,Fr+)

= soluble sodium fluoride, NaF, is soluble

any anion + hydrogen ion

[H+(aq)]

= soluble hydrogen chloride, HCl, is soluble

any anion + amonium ion

(NH4+)

= soluble ammonium chloride, NH4Cl, is soluble

nitrate

NO3-

+ any cation = soluble potassium nitrate, KNO3, is soluble

acetate

(CH3COO-)

+ any cation = soluble sodium acetate, CH3COONa, is soluble

Chloride (Cl-),

Bromide (Br-

),Iodide (I-)

+ silver (Ag+), lead (Pb2+), mercury (Hg2+), copper

(Cu+

), thallium (Tl+

)

= low solubility (insoluble) silver chloride, AgCl, forms a white

precipitate (a white solid)

+ any other cation = soluble potassium bromide, KBr, is soluble

Sulphate

(SO42-)

+ calcium (Ca2+), strontium (Sr2+), barium (Ba2+), silver

(Ag+), lead (Pb2+), radium (Ra2+)

= low solubility (insoluble) barium sulphate, BaSO4, forms a white

precipitate (a white solid)

+ any other cation = soluble copper sulphate, CuSO4, is soluble

Sulfide

S2-

+ alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+), alkali earth

metals (Be2+,Mg2+,Ca2+,Sr2+,Ba2+,Ra2+),

and H+(aq), NH4+

= soluble magnesium sulfide, MgS, is soluble

+ any other cation = low solubility (insoluble) zinc sulfide, ZnS, is insoluble

Hydroxide

OH-

+ alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+),

H+(aq),NH4+,Sr2+,Ba2+,Ra2+,Tl+

= soluble strontium hydroxide,

Sr(OH)2, is soluble

+ any other cation = low solubility (insoluble) silver hydroxide, AgOH, is insoluble

(forms a precipitate)

Phosphate, PO43-,

Carbonate, CO32-,

l hi SO 2

+ alkali ions (Li+,Na+,K+,Rb+,Cs+,Fr+), H+(aq),NH4+ = soluble ammonium phosphate,

(NH4)3PO4, is soluble

h i l l bili (i l bl ) i b M CO i

Anwar Slide Presentation

Documents

Transcript of Anwar Slide Presentation