phycoremediation of artificial bathroom greywater … · sumber-sumber pencemaran air di malaysia...
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PHYCOREMEDIATION OF ARTIFICIAL BATHROOM
GREYWATER IN VILLAGE HOUSES USING MICROALGAE
Botryococcus sp.
ANWARUDDIN AHMED WUROCHEKKE
A thesis submitted in
Fulfillment of the requirement for the award of the
Degree of Doctor of Philosophy of Civil and Environmental Engineering
Faculty of Civil and Environmental Engineering
UNIVERSITI TUN HUSSEIN ONN MALAYSIA
86400 Parit Raja, Johor
MARCH, 2016
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DEDICATION
I dedicate this work to my beloved son Ahmed Rayyan and my darling wife Hauwa
Atiku.
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ACKNOWLEDGEMENT
First of all most importantly, Allah, the all Mighty, who gave me the strength and
courage to carry-on throughout my studies, making it all possible.
I wish to express my sincere appreciation and gratitude to the following persons and
institution for their contributions to the successful completion of this study. My candid
appreciation to my supervisor Dr. Radin Maya Saphira Radin Mohamed for her
ceaseless encouragement, support, advice, tolerance. She is not just a supervisor but a
dear sister and a times a mother, my gratitude please, only Allah can pay you back.
My appreciation goes to my co-supervisors Prof. Ir. Dr. Amir Hashim Mohd. Kassim
and Dr. Hazel Monica Matias-Peralta thank you. The staff of wastewater, analytical
laboratory FKAAS, ceramic laboratory FKMP and UTHM with sponsorship under
grant incentive scheme (GIPS), postgraduate scholarship and fundamental research
grant scheme (FRGS) thanks a lot (Terima Kasih Banya Banya).
My sincere appreciation to my wife Hauwa Atiku with her resolute, patience,
perseverance and tolerance. To my son Ahmed Rayyan am sorry, daddy is always not
with you (crying on 14/12/20016 at 8 pm my daddy is not back home yet). Daddy will
never let you go down dear, I love you.
My heartfelt gratitude goes to my dad Alhaji Ahmed Usman Wurochekke, mum Khadija
Ahmed and siblings. My in-law Air commodore Atiku Umar (Rtd) and family, my Aunties
especially Halima and Safiya Modibbo. My uncle Mahmud Usman, Isa Modibbo. Next is
my brother Group Captain Shehu Bakari, Mr, Mohammed Giddado Modibbo and Huzaifa
Ahmed, thank you for always praying, loving, providing and always being there for me.
Lastly, a big thank you to my team in UTHM Suriani, Wahida, Hasila, Aznin, Azimah,
other UTHM collique, Dr. Adel Algeethi and my grandma, friends, brothers, sisters,
nieces and nephews, I love you all.
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ABSTRACT
The sources of water pollution in Malaysia are domestic sewage and industrial waste.
Direct discharge of household bathroom greywater into drains cause euthrophication
into the water bodies. Phycoremediation of bathroom greywater effluent to meet a
certain level of discharge limit using microalgae Botryococcus sp. is suggested. The
objectives of this study is to asses quality of nutrients in raw bathroom greywater,
produce artificial bathroom greywater (ABGW) recipe with Response Surface
Methodology, to study biokinetic absorption of microalgae through phycoremediation.
To optimize Botryococcus sp. cell concentration, pH and the efficiency of laboratory
scale treatment system with Botryococcus sp. was observed. The first objective results
shown that NO3-N and PO4-P were 1.03-7.54 & 0.12-22.7 mg/L respectively and 63
L/c/day was discharged to drains as raw bathroom greywater. Secondly, ABGW recipe
for soap, detergent, shampoo, shower gel, toothpaste were 0.13, 0.97, 0.88, 0.34, 0.37
mg/L respectively and pH= 6.55. The optimum concentration of Botryococcus sp. was
106 cells/mL and pH 7 for the third objective. Fourthly, the efficiency of Botryococcus
sp. in removing NO3-N was 97% and PO4-P 87% in ABGW on the 30th day of
phycoremediation, while biokinetic absorption rate using Michaelis-Menten
coefficient were K =0.46 mgNO3-N mg/chl a/day & mK =12.501 mg/L (R2 = 0.83)
and PO4-P coefficients were K =8.53 mgPO4-P mg/chl a/day & mK =176.88 mg/L (R2
= 0.94). Lastly, the efficiency of Botryococcus sp. in laboratory scale treatment system
was 90.98% and 93.88% for NO3-N while 80.9% and 83% for PO4-P on the 13th day
of phycoremediation in ABGW and raw bathroom greywater respectively.
Statistically, algal days of culture, growth of algae, pH, temperature and light
correlated well (p<0.05 & 0.01) influencing high nutrient removal in the system.
Therefore, this proves that Botryococcus sp. has high potential to absorb NO3-N and
PO4-P from household bathroom greywater. Hence, the system of this study represents
an effective solution for remediation of bathroom greywater.
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ABSTRAK
Sumber-sumber pencemaran air di Malaysia adalah sisa kumbahan domestik dan sisa
industri. Pelepasan langsung air sisa dari bilik air rumah kediaman ke dalam longkang
menyebabkan berlakunya euthrophikasi ke dalam saliran air semulajadi. Penggunaan
mikroalga Botryococcus sp. bagi pykoremediasi air sisa dari bilik mandi bagi
memenuhi tahap tertentu had pelepasan adalah dicadangkan. Objektif kajian ini adalah
untuk menilai kualiti nutrien dalam air sisa dari bilik mandi, menghasilkan resepi air
sisa dari bilik mandi buatan (ABGW) dengan Kaedah Response Surface, untuk
mengkaji penyerapan biokinetic mikroalga melalui pykoremediasi. Bagi
mengoptimumkan kepekatan sel Botryococcus sp., pH dan kecekapan sistem rawatan
skala makmal dengan Botryococcus sp. diperhatikan. Bagi objektif pertama, kajian
menunjukkan bahawa NO3-N dan PO4-P bagi air sisa bilik mandi masing-masing
mempunyai nilai 1.03-7.54 & 0.12-22.7 mg/L dan 63 L/c/ hari telah dilepaskan ke
longkang. Kedua, resipi ABGW untuk sabun, bahan pencuci, syampu, gel mandian,
ubat gigi masing-masing adalah 0.13, 0.97, 0.88, 0.34, 0.37 mg/L dan pH = 6.55.
Kepekatan optimum Botryococcus sp. adalah 106 sel / mL dan pH 7 untuk objektif
ketiga. Keempat, kecekapan Botryococcus sp. dalam menghapuskan NO3-N adalah
97% dan PO4-P 87% pada ABGW pada hari ke-30 pykoremediasi, manakala kadar
penyerapan biokinetic menggunakan pekali Michaelis-Menten adalah K = 0.46
mgNO3-N mg/chl a/hari & mK = 12.501 mg/L (R2 = 0.83) dan PO4-P pekali adalah
K = 8.53 mgPO4-P mg/chl a/hari & mK = 176.88 mg/L (R2 = 0.94). Akhir sekali,
kecekapan Botryococcus sp. dalam sistem rawatan skala makmal bagi NO3-N adalah
90.98% dan 93.88% manakala 80.9% dan 83% untuk PO4-P pada hari ke-13 dalam
pykoredemiasi ABGW dan air sisa dari bilik mandi yang belum diproses. Secara
statistik, hari pembiakan alga, kadart pertumbuhan alga, pH, suhu dan cahaya
mempunyai kaitan yang baik (p <0.05 & 0.01) mempengaruhi penyingkiran nutrien
yang tinggi di dalam sistem. Oleh itu, ini membuktikan bahawa Botryococcus sp.
mempunyai potensi yang tinggi untuk menyerap NO3-N dan PO4-P dari air sisa dari
bilik air rumah kediaman. Oleh itu, sistem kajian ini merupakan penyelesaian yang
berkesan untuk pemulihan air sisa mandian.
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TABLE OF CONTENTS
DECELERATION ii
DEDICATION iii
ACKNOWLEDGMENT iv
ABSTRACT v
ABSTRAK vi
TABLE OF CONTENTS vii
LIST OF TABLES ix
LIST OF FIGURES x
LIST OF APPENDICES xi
LIST OF ABBREVIATIONS xii
LIST OF PUBLICATIONS xiii
CHAPTER 1 INTRODUCTION 1
1.1 Background 1
1.2 Problem Statement 3
1.3 Aim of study 4
1.4 Objective of study 4
1.5 Scope of Study 5
1.6 Hypothesis of Research 6
1.7 Significance of Study 6
CHAPTER 2 LITERATURE REVIEW 8
2.1 Introduction 8
2.2 Composition of Sources and Characteristics of
Greywater 9
2.3 Greywater Production 12
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2.4 Standard Regulations 15
2.5 Response Surface Methodology (RSM) 17
2.6 Artificial Greywater 17
2.7 The Trend of Greywater Treatment 19
2.7.1 The Use of Natural Filter Material in
Greywater Treatment 19
2.8 Potential Use of Microalgae in Treating Greywater 24
2.9 Microalgae Botryococcus sp. 25
2.10 Phycoremediation by using Microalgae 26
2.11 Growth Conditions of Microalgae (Botryococcus sp.) 32
2.12 Growth Controlling Factors of Microalgae 32
2.12.1 Nitrogen and Phosphorus 33
2.12.2 Carbon 33
2.12.3 Light Intensity 33
2.12.4 Temperature 34
2.12.5 pH 34
2.13 Nitrogen, Phosphorus and the Environment 35
2.14 Nutrients Removal for Total Nitrogen and Total
Phosphorus 37
2.15 Mechanisms of Nutrient Removal 37
2.15.1 Nitrogen 38
2.15.2 Phosphorus 41
2.16 Biokinetic Studies on Phycoremediation of
Microalgae 43
2.17 Selection of Greywater Treatment Method 45
2.18 Photobioreactors during the Phycoremediation
Process 47
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2.19 Key Findings of the Literature Review 47
CHAPTER 3 METHODOLOGY 49
3.1 Introduction 49
3.2 Methodology Flowchart 50
3.3 Site selection, Survey and Sampling 51
3.4 Characteristics of Raw Bathroom Greywater
Samples 54
3.4.1 Preservation and Storage 54
3.4.2 Bathroom Greywater Production and
Loading Rates from Four Houses 55
3.5 Optimization using Response Surface
Methodology for Artificial Bathroom Greywater
(ABGW) Recipe Production 56
3.6 Microalgae Botryococcus sp. Morphological and
Molecular Identification 57
3.7 Culturing Microalgae Botryococcus sp. 59
3.7.1 Algal Cell Count and Inoculation 61
3.8 Optimization of pH and Cell Concentration of
Microalgae Botryococcus sp. 61
3.9 Phycoremediation of Bathroom Greywater 62
3.9.1 Biokinetic Absorption Rate of
Microalgae Botryococcus sp. 63
3.10 Laboratory Scale Greywater Treatment System
Set-up for Bathroom Greywater 64
3.10.1 Statistical Analysis of Environmental Factor
Influence on Botryococcus sp. in Laboratory
Scale Greywater Treatment System 65
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CHAPTER 4 RESULT AND DISCUSSION 66
4.1 Introduction 66
4.2 Household Activity for Bathroom Greywater 67
4.2.1 Bathroom Greywater Production from
Household Case Study 69
4.2.2 Bathroom Greywater Pollutant Loading Rate 71
4.3 Characteristics of Bathroom Greywater 72
4.4 Optimization of Soap, Detergent, Toothpaste,
Shampoo, Shower gel and pH in the Formulation
of Artificial Bathroom Greywater 76
4.4.1 Optimization Experiments for Artificial
Bathroom Greywater (ABGW) Recipe
Formulation 78
4.5 Artificial Bathroom Greywater (ABGW)
Recipe Characteristics 79
4.6 Morphological Characteristics and Molecular
Identification of Microalgae Botryococcus sp. 81
4.7 Optimization of Cell Concentration for
Microalgae Botryococcus sp. in Artificial
Bathroom Greywater (ABGW) 82
4.8 pH Optimization of Botryococcus sp. 87
4.9 Nutrient (Nitrate-N and Phosphate-P) Removal
Efficiency by Botryococcus sp. from Artificial
Bathroom Greywater (ABGW) 88
4.9.1 Biokinetic Coefficients, Nitrate-N and
Phosphate-P Specific Removal Rate and
Yield Coefficients from Artificial
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Bathroom Greywater (ABGW) 93
4.10 The Efficiency of Botryococcus sp. in Laboratory
Scale Photobioreactor Treatment System for
Nutrient Removal from Raw and Artificial
Bathroom Greywater (ABGW) 99
4.10.1 Statistical Analysis for Environmental
Factors on Botryococcus sp. Growth 106
4.11 Summary 107
CHAPTER 5 CONCLUSION AND RECOMMENDATION 109
5.1 Conclusion 109
5.2 Recommendation 111
REFERENCES 113
APPENDIX A Molecular Identification of Microalagae
Botryococcus sp. Results 130
APPENDIX B SAMPLE OF SURVEY QUESTIONAIRE 134
APPENDIX C RSM RESULTS 137
VITA 146
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LIST OF TABLES
2. 1: Characteristics of Bathroom Greywater and Related
Literature Sources 11
2. 2: Quantity of Greywater Produced from Shower and
Bathroom Sources Compared with Literature Data 14
2. 3: Effluent Standards in Selected Countries for
Discharge into Surface Water (Alderlieste et al., 2006) 16
2. 4: Environment Quality (Sewerage and Industrial
Effluents) Regulations, 2009 (Environment Quality
Act 1974) 16
2. 5: Artificial Greywater Recipe 18
2. 6: Review of Primary Treatment System of
Different Sources of Greywater 21
2. 7: Reduction of N and P by using Microalgae in
Treating Different Types of Wastewater 30
3. 1: Characteristics Measurement of Bathroom Greywater
Parameters, Equipment’s and Models 54
3. 2: Preservation and storage of water samples
(APHA, 2012) 55
3. 3: Optimization of factor dosage and pH range using
design
expert for artificial bathroom greywater recipe 57
3. 4: Composition of Bold’s Basal Medium for Culturing of
Botryococcus sp. 60
3. 5: Bathroom greywater produce per person per day 64
4. 1: Summary of Demographic Data, Type of Bath,
Household Activities and Bathroom Greywater
Production Profile 68
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4. 2: Bathroom greywater production rate (L/peak
period/day) 70
4. 3: Bathroom Greywater BOD5 Loading Rate of
Bathroom Greywater at House A, B, C and D (n=5) 72
4.4: Range of Raw Bathroom Greywater Characteristics
of Houses A, B, C and D (Sampling was done from
March 2014 to February 2015, from 6-9 in the morning
and 5-8 at night, n=15) 73
4. 5: Range of Characteristics from Raw Bathroom
Greywater Compared with Previous Studies 74
4. 6: Model and Experimental Values for the Optimization
Experiments 79
4. 7: Comparison of Artificial and Raw Bathroom
Greywater Characteristics with Previous
Studies (n=5). 80
4. 8: Temperature and Growth of Botryococcus sp.
for Raw Bathroom Greywater and Artificial
Bathroom Greywater (ABGW) 103
4. 9: Correlation Analysis for Environmental Factors
on Algal Growth 107
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LIST OF FIGURES
2. 1: Nitrogen Cycle (Stanley, 2001) 36
2. 2: Conversion of inorganic nitrogen to organic nitrogen
(assimilation) (Cai et al., 2013). 38
3. 1: Frame work of methodology 50
3. 2: Satellite image of sampling area (coordinate: 1° 54’ 0”
North, 103° 9’ 0” East), Parit Raja, Batu pahat, Johor,
Malaysia (source: Google Maps) 51
3. 3: Bathroom greywater discharge from four houses
sampling point (Photos were taken on 20/07/2015) 53
3. 4: Image of Microalgae Botryococcus sp. Observed under
Light Compound Microscope 58
3. 5: (a) Mixture of BBM Stock Solution without Microalgae
Botryococcus sp. (b) Culturing Process Inoculated
Microalgae Botryococcus sp. (Photo was taken on
28/5/2015). 60
3. 6: (a) Multi Parameter Meter, model: DR1900 HACH
(b) Ion Chromatography (IC), Dionex, model: ICS-2000
(c) UV-VIS Spectrophotometer, model: DR 6000 HACH
(Photo was taken 9/4/2015) 63
3. 7: Bathroom Greywater Laboratory Scale Greywater
Treatment System (Photobioreactor) (Photo was taken
on 12/10/2014) 65
4. 1: Percentage of Bathroom Greywater Production from
the Four Houses in the Case Study 70
4. 2: Botryococcus sp. Cell Diameters 82
xv
4. 3: Botryococcus sp. Initial Inoculum of 105 Cell/mL in
ABGW 83
4. 4: Botryococcus sp. Initial Inoculum of 106 Cell/mL in
ABGW 84
4. 5: Botryococcus sp. Initial Inoculum of 107 Cell/mL in
ABGW 85
4. 6: Optimum Specific Growth Rate of Botryococcus sp. in
ABGW, BBM (+) Control and Distilled water
(-) Control 86
4. 7: Specific Growth Rate of Botryococcus sp. for Varying
Concentrations of ABGW 86
4. 8: Cell Growth of Botryococcus sp. against time for
Optimum pH 88
4. 9: ABGW Nitrate-N Removal Concentration and
Removal Percentage 89
4. 10: ABGW Phosphate-P Removal Concentration and
Removal Percentage 91
4. 11: Variation of chl a Content with initial Nitrate-N
Concentration 92
4: 12. Effect of Initial NNO 3 on Specific NNO 3
Removal Rate 94
4. 13: Effects of Initial PPO 4 on Specific PPO 4
Removal Rate 94
4. 14: Determination of Kinetic Coefficients, mK and K for
NNO 3 Removal 96
4. 15: Determination of Kinetic Coefficients, mK and K for
PPO 4 Removal 96
4. 16: Determination of Yield Coefficient for NNO 3
Uptake by Botryococcus sp. 97
4. 17: Determination of Yield Coefficient for PPO 4
Uptake by Botryococcus sp. 97
xvi
4. 18: Nitrate-N Concentration and Percentage of Removal
against Time 100
4. 19: DO Profiles for Raw Bathroom Greywater and
Artificial Bathroom Greywater (ABGW) against Time 102
4.20: Phosphate-P Concentration and Percentage of
Removal against Time 104
xvii
LIST OF ABBREVIATIONS
BOD - Biochemical Oxygen Demand
COD - Chemical Oxygen Demand
TSS - Total Suspended Solids
NH4+ - Ammonium
NO3- - Nitrate
NO3-N - Nitrate-N
PO4-P - Phosphate-P
OP43- - Orthophosphate
CO2 - Carbondioxide
DHA - Docosahexaenoic Acid
GHG - Green House Gases
Mg - Magnesium
Ca - Calcium
TN - Total Nitrogen
N - Nitrogen
P - Phosphorus
TP - Total Phosphorus
DNA - Dioxoribonucleic Acid
RNA - Ribonucleic Acid
ABGW- Artificial Bathroom Greywater
ATP - Adesonine triphosphate
RSM - Response Surface Methodology
PDDA - Polyelectrolyte polydiallyldimethyl Ammonium Chloride
DAF - Dissolved Air Flotation
CTAB - Cationic N-cetyl-N-N-N trimethyl Ammonium
NaCl - Sodium Chloride
UTHM - Universiti Tun Hussein Onn Malaysia
xviii
APHA - American public health association
CCD - Central Composite Design
SMBR - Submerged membrane bioreactor
FSTPI - Fakulti Sains, Teknology dan Pembangunan Insan
xix
LIST OF PUBLICATIONS
Anwaruddin Ahmed Wurochekke, Radin Maya Saphira Radin Mohamed, Amir
Hashim Mohd. Kassim. Adel Algeethi, Hauwa Atiku, Hazel Monica Matias-Peralta.
2016. Household Greywater Treatment Methods using Natural Materials and their
Hybrid System: A Review, Journal of Water and Health, 14(6), 914-928 (ISI, IF=
1.025, Q2).
Hauwa Atiku Majidda, Radin Maya Saphira Radin Mohamed, Al-Gheethi AA,
Anwaruddin Ahmed Wurochekke & Amir Hashim Mohd Kassim. 2016. Harvesting
Microalgae Biomass from the Phycoremediation Process of Greywater. Environmental
Science and Pollution Research, 23(16) (ISI, IF= 2.8, Q1).
Anwaruddin Ahmed Wurochekke, Radin Maya Saphira Radin Mohamed, Siti
Asmah Binti Lokman Halim, Amir Hashim bin Mohd. Kassim and Rafidah Binti
Hamdan. 2015. Sustainable Extensive On-Site Constructed Wetland for Some
Bacteriological Reduction in Kitchen Greywater. In Applied Mechanics and
Materials (773), 1199-1204) (Scopus).
Anwaruddin Ahmed Wurochekke, Radin Maya Saphira Radin Mohamed, Chee-
Ming Chan and Amir Hashim bin Mohd. Kassim. 2014. The Use of Natural Filter
Media Added with Peat Soil for Household Greywater Treatment. International
Journal of Engineering Technology (JET) 2(4).
Radin Maya Saphira Radin Mohamed, Anwaruddin Ahmed Wurochekke, Siti
Solehah Mohd Hadzri and Amir Hashim Mohd Kassim. 2015. Induction Performance
of pn-Site Low Cost Treatment Unit for Treating Kitchen Greywater at Village House.
Journal of Applied Sciences Research 11(10), 22-28.
xx
Anwaruddin Ahmed Wurochekke, Nurul Azma Harun, Radin Maya Saphira Radin
Mohamed and Amir Hashim Bin Mohd. Kassim, (2014). Constructed Wetland of
Lepironia Articulata for Household Greywater Treatment presented at the
INTERNATIONAL CONFERENCE ON ENVIRONMENTAL SCIENCE AND
DEVELOPEMENT Singapore, 20th February, 2014.
Radin Maya Saphira binti Radin Mohamed, Anwaruddin Ahmed Wurochekke, Siti
Solehah binti Mohd Hadzri, Sabariah Musa and Amir Hashim bin Mohd. Kassim,
(2014). Preliminary Performance of On-Site Low Cost Treatment Unit for Treating
Kitchen Greywater At Village House presented at the NATIONAL SEMINAR ON
CIVIL ENGINEERING RESEARCH SEPKA, Johor, 14th April, 2014.
Anwaruddin Ahmed Wurochekke, Radin Maya Saphira Radin Mohamed, Hauwa
Atiku, Suriani Sharuddin and Amir Hashim Mohd. Kassim. Improvement of Bathroom
Greywater Quality after Phycoremediation with Microalgae Botryococcus sp.
presented at the INTERNATIONAL CONFERENCE ON ENVIRONMENTAL
FORENSICS (iENFORCE 2015), 19-20, August 2015.
Hauwa Atiku, Radin Maya Saphira Radin Mohamed and Anwaruddin Ahmed
Wurochekke. Bathroom Greywater Bioremediation by Microalgae Botryococcus sp.
(ICSESS 2016) presented at the 2nd INTERNATIONAL CONFERENCE ON
SCIENCE, ENGINEERING AND THE SOCIAL SCIENCES, Universiti Teknologi
Malaysia, Johor Bahru, Malaysia. Organized by UTM International and UTHM, 29
May-1 June 2016.
Al-Gheethi AA, Mohamed RMSR, Wurochekke AA, Nurulainee NR, Mas Rahayu J,
Amir Hashim MK. Efficiency of Moringa oleifera seeds for Treatment of Laundry
Wastewater presented at the INTERNATIONAL SYMPOSIUM ON CIVIL AND
ENVIRONMENTAL ENGINEERING (ISCEE 2016), 25-26 October, 2016.
1
CHAPTER 1
INTRODUCTION
1.1 Background
The speedy growth of the human population and their activities have caused serious
water pollution and insufficient freshwater in many developing countries. It is a
driving force for everybody to think of alternatives and sustainable solutions to manage
this valuable resource (Qu et al., 2013). In Jordan, water scarcity occurred due to the
growing population over the previous decade. The availability per capita of water
decreased to 198 m3/capita/year from the standard of 1000 m3/capita/year from 1996
to 2013 (Boufaroua et al., 2013; Najib Al-Beiruti, 2005) which has affected the
economic growth in Jordan. The high cost of usual sewers is one of the main restraints
to expand wastewater services to small communities. As a result, domestic wastewater
especially from greywater sources goes uncollected contributing phosphorus, nitrogen
and other elements into water bodies. However, Jordanian water management has
implemented some models to change how water is handled and appreciated due to the
present water plan from the ministry of water and irrigation which is responsible for
the management of operations, maintenance cost and supply, treating and distribution
(Al-Beiruti, 2007). Hence, these would increase greywater reuse and reduce the
amount of pollutants being discharged into water bodies.
In India, about 25 billion liters of untreated wastewater are discharged into the water
bodies daily (Parjane & Sane, 2011). This untreated wastewater will lead to health
problems and increase infectious diseases such as diarrhea, dysentery, skin and tissue
2
infections (Akpor et al., 2011). Furthermore, in India, the International Water
Management Institute (IWMI) forecast that by 2025 one person in three will facetotal
water shortage. Similarly, rapid development and industrialization in Malaysia have
affected water resources especially in the rural community. Besides, the decrease in
quality of water in many rivers has mainly contributed to the water problem in
Malaysia. The rivers of Kuantan and Belat were monitored by Kozaki et al., (2016)
and it was found that the pollution level of some ions were as follows: 4.44-14.9 for
4NH , 3.71-18.1 for
3NO ,154-6429 for Na and 33.8-1363 for 2Mg respectively.
These pollutants were influenced by human activities like the discharge of untreated
water from human sewage and household waste, and also industrial point sources.
Thus, they are becoming serious issues and need to be solved before worsening.
Greywater is another alternative source to substitute freshwater usage but it contains
contaminants (Parjane & Sane, 2011). Greywater is a type of wastewater derived from
the kitchen, bathroom (i.e. discharge from the hand basin, shower, and bath) and also
laundry water. Bathroom contributes more than 50% of the total usable greywater
volume in a typical household (Donner et al., 2010; Coghlan & Higgs, 2003). Besides
that, greywater which originates from bathrooms and showers make up over 30 per
cent of household greywater flow (Edwin et al., 2014). Water used for washing hands
and showers generate about 50-60% of the total greywater and it can be considered as
least polluted type of greywater compared to others. Common chemical pollutants
include soap, shampoo, hair dye, toothpaste and cleaning products. It also has a
number of faecal contaminations 3.2 x 105 CFU/100mL through body wash (Saumya
et al., 2015).
However, greywater does not include the wastewater that is generated from toilet use
which is normally considered as black water. However, greywater is generated in
different quantities between household to household within one community and
depends on different factors such as lifestyle and household activities (Al-Mashaqbeh
et al., 2012). Many communities in Malaysia, mainly villagers, dispose bathroom
greywater directly into the nearest ditch. These inputs promote eutrophication in water
bodies and naturally there will be a decline in water quality. At the same time, the
removal of nutrients and organic compounds from greywater is an essential means to
3
avert eutrophication and water bloom. Therefore, bathroom greywater ought to obtain
proper treatment prior to being discharged into water bodies.
Numerous systems operate for the removal of nutrients from greywater, although these
are expensive and generate elevated thick soft mud (Ruiz-Marin et al., 2010). Natural
treatment systems via primary settling with cascaded water flow, aeration, agitation
and filtration (Parjane & Sane, 2011; Pangarkar et al., 2010) were used and are less
expensive. Yet, there is lack of information when it concerns the removal of nutrients
especially phycoremediation with microalgae Botryococcus sp. Greywater
management treatment system is a system that allows direct utilization of the water. It
can use the natural gravity by a hybrid treatment process with the use of natural
materials and wetland system. It will facilitate in breaking down the organic
compounds and recovery of nutrients (Parjane & Sane, 2011). Bathroom greywater
should preferably be treated anaerobically because of lower treatment costs and the
possibility of recovering energy (Leal et al., 2010).
Therefore, in this study phycoremediation with microalgae Botryococcus sp. as a
laboratory scale system will be adopted. Based on the aforementioned criteria,
greywater treatment options for households will be advocated. Since uptake is the
major means of removing nutrients by microalgae, the colony of microalgal growth
squarely influences the nutrient removal rate. Greywater can contain nutrients such as
total phosphorus (TP), total nitrogen (TN) from detergents (Maya et al., 2013b; Park
et al., 2011b) and total organic carbon (TOC) (Beck et al., 2013b; Li et al., 2009) that
benefit algal growth. Therefore, the usage of microalgae for treatment was proposed
for a simple greywater treatment system especially for greywater from the bathroom
source.
1.2 Problem Statement
Conventional discharge of greywater into drains gains the least attention in terms of
environmental sanitation such as toilet waste and solid waste disposals. In individual
village house areas in Malaysia, bathroom greywater is most often discharged
untreated into storm water drains. These discharge can cause unpleasant odours, bread
4
mosquitos, flies, aesthetic of the environment is disturb and add nutrients (nitrogen
and phosphorus) in the drain. Excess nutrients in bathroom greywater causes
eutrophication. This discharge of untreated bathroom greywater in an uncontrolled
manner to the main drain with excess nutrients flow in to the rivers. Rivers are the
main source of water supply in Malaysia. Therefore, eutrophication of rivers with
enrich nutrients rapidly grow algae. The decaying algae plant reduce dissolved oxygen,
fish and other aquatic life die. In addition, the rapid increase of pollutants (nutrients
and pathogens) will occur actively and result in an unhealthy environment for humans
or animals with dangerous diseases. Bathroom greywater effluents have high
concentrations of nitrogen and phosphorus content which are normally generated by
activities such as bathing. It is anticipated that the utmost source of nitrogen is urine,
as some people pass urine in the shower rooms or from body washing and bathing
through the use of protein-rich shampoos and conditioners. Additionally, nutrient
loads may come from washing babies, children after defecation, diaper changes or
diaper washing. Phosphorus is mainly found in detergents. Therefore, bathroom
greywater treatment is necessary before discharging the treated greywater to water
bodies.
1.3 Aim of study
The aim of this study is to evaluate the efficiency of phycoremediation in treating
bathroom greywater from village households. This study intends to ascertain the
potential for treated bathroom greywater to reduce pollutant loads of nearby water
bodies.
1.4 Objective of study
This study embarks on the following objectives:
1. To assess the nutrient, elements availability and the quality and quantity of
bathroom greywater from selected case study houses.
2. To optimize bathroom products for the production of an artificial bathroom
greywater (ABGW) recipe.
5
3. To optimize Botryococcus sp. cell concentration for best growth and pH of
ABGW.
4. To determine ABGW nutrient removal efficiency and mechanism on biokinetic
absorption rate.
5. To determine the efficiency of Laboratory Scale Greywater Treatment System
for nutrient (nitrate-N and phosphate-P) removal and influence of
environmental factors during phycoremediation of ABGW and raw bathroom
greywater in a photobioreactor using Botryococcus sp.
1.5 Scope of Study
The scope of this study includes the isolation of a microalgae called Botryococcus sp.
at the microbiology laboratory, Fakulti Sains, Teknology dan Pembangunan Insan (FSTPi),
Universiti Tun Hussein Onn Malaysia (UTHM). This microalgae was originally
obtained from the Endau Rompin forest reserve in Johor, Malaysia.
Household activities were ascertained using interviews and a questionnaire in four
households around Parit Raja. Bathroom greywater loading was measured during the
composite sampling by comparing the production of bathroom greywater in the
mornings and in the night. Water quality parameters were tested for pH, BOD5, COD,
TSS, Turbidity, ammonium, nitrate-N, phosphate-P, Ca, Na and Mg to assess the
quality of bathroom greywater collected. The artificial bathroom greywater recipe was
produced using domestic products. Phycoremediation was done on bathroom
greywater with Botryococcus sp. to determine its efficiency on nutrient removal
(nitrate-N and phosphate-P) and chloropyll-a content. The adsorption rate of
Botryococcus sp. on nitrate-N and phosphate-P were determined with a biokinetic
model of Michaealis-Menten equation o
m
xi SK
K
KR
111 and environmental factors
like light, temperature and pH on algal growth were observed during
phycoremediation.
The Laboratory Scale Greywater Treatment System (photobioreactor) was built to
represent the phycoremediation efficiency of ABGW and raw bathroom greywater for
6
village household application to reduce the pollution load before entering the main
river. It is a unit named photobioreactor which contains the microalgae Botryococcus
sp. Optimization of the microalgae cell concentration and pH for Botryococcus sp.
growth were conducted. Optimization of simulated ABGW was also done using
Response Surface Methodology (RSM). The efficiency of Laboratory Scale Greywater
Treatment System was determined. Statistical analysis on environmental factors was
run to determine their influence on the growth of Botryococcus sp. Recommendations
based on the efficiency were outlined.
1.6 Hypothesis of Research
This study hypothesized that nutrients supplied from bathroom greywater can be
absorbed by microalgae for growth. However, it is expected that the growth of
microalgae is also influenced by pH, temperature and light supply. Some chemicals
from soaps, shampoos, household cleaning products may affect the microalgae growth.
Yet, the composition and amount of nutrients, the variation of household activities and
the effects of environmental parameters on the growth of microalgae are not well
understood. Effluent treatment is expected to meet a certain level of discharge limit,
therefore phycoremediation can reduce the pollution load of bathroom greywater
effluent.
1.7 Significance of Study
The most important contribution of this study is to produce a bathroom greywater
treatment system by capturing the nutrients for microalgae growth during the
phycoremediation process. The phycoremediation system is a treatment system that
can be fit in an individual house that could serve as a greywater pollutant remover.
This can reduce the pollution load especially nutrients to an acceptable standard limit
to be discharged into water bodies. Through phycoremediation, the microalgae tends
to trap CO2 from the atmosphere during photosynthesis thus it can control the
production of greenhouse gases and oxygenate the environment.
7
Therefore, Malaysia’s water resource authorities will benefit from the outcome of this
research with greywater management strategies. In Malaysia, there is no definite
standard of household greywater. The Environmental Quality (Sewage and Industry
Effluences) Regulations 1979 are used to refer to the discharge of effluence into any
inland water other than the effluence discharged from prescribed premises. Hence, this
study will initiate the development of a national guiding principle for the treatment of
household greywater to prevent the contamination of the environment.
8
CHAPTER 2
LITERATURE REVIEW
2.1 Introduction
Greywater is defined as wastewater generated from domestic activities such as dish
washing, laundry and bathing as well as wastewater disposed by kitchen sinks (Oron
et al., 2014). Greywater has less organic loading than that of sewage (blackwater).
Some authors in the literature categorize kitchen wastewater as blackwater due to its
high organic loading compared to other sources of wastewater such as bath water.
However, blackwater might be used to indicate the source of high organic contents
which is associated with human wastes and thus has low quality and high microbial
loads (Allen et al., 2010).
When untreated greywater is discharged into the river, the quality of freshwater
deteriorates. Untreated greywater is also one of the major contributors for
eutrophication which reduces portable water for human consumption. This situation
occurs where the water body receives an excessive amount of nutrients that causes
negative impacts on the environment such as the reduction of oxygen levels in the air
as well as a decrease in the species of fish and other microbe populations (Gera et al.,
2015). A review on the bathroom greywater characteristics, the trends of treatment
systems for bathroom greywater and their related sources, the potential of
phycoremediation by using photobioreactors, nutrient removal and their mechanisms
will be discussed in this chapter.
9
2.2 Composition of Sources and Characteristics of Greywater
Greywater represents the largest potential source of water saving in domestic
residences, accounting for as much as 50-80% of the total water use (Al-Hamaiedeh
& Bino, 2010). Greywater is sourced from laundries, hand basins, bathrooms and
kitchens.
Greywater from showers normally contains soaps, shampoos, body-fats, hair, fabric
fibre, urine and skin. The latter is more prevalent where the family comprises either
very young children or the incontinent elderly. In addition, greywater may contain
household cleaning products (Christova-Boal et al., 1996). There are three
characteristics of greywater which are physical, chemical and biological.
Characteristics of greywater depends on the quality of water, the distribution of both
clean drinking water and greywater leaching from pipes, chemical and biological
processes in the biofilm on the pipe wall and the activities performed by the household.
Physical characteristics of greywater are primarily and generally measurable
substances such as pH, alkalinity, conductivity, suspended solids (SS), turbidity and
color. The age of greywater is identified by its appearance (colour). Also, temperature
plays a vital role in the determination of those parameters. Greywater is generally low
in suspended solids as it is collected from laundry and bathroom sources. Shaving
waste, skin, hair and lint are common suspended solids which can potentially foul
treatment processes.
The chemical compound representatives from households are from cooking and
washing. The amount and variability of the chemical compounds are affected by water
usage by individual habits, fixtures, product use (e.g. detergents, shampoos, soaps etc.)
and other site-specific characteristics. In addition, the amount of salt (sodium, calcium,
magnesium, potassium and other salt compounds), oils and grease can largely be
influenced by the type of products used within a household (Mohamed et al., 2014).
Organic and inorganic compounds found in greywater evidently prove the existence
of chemical compounds like synthetic organic chemicals, ammonia, phosphorus,
carbon, hydrogen, oxygen and trace elements such as sulfur.
10
Biological parameters are perhaps of greatest importance to greywater treatment. The
microbiological compound originates from the presence of potential harmful
microorganisms. These are feacal coliforms and E. coli bacteria in greywater
pollutants (Eriksson et al., 2002). The feacal coliform bacteria are used as an indicator
of microbiological quality. These bacteria can pose a risk to human health where an
illness or infection may occur through contact with greywater.
The general characteristics of raw household wastewater are summarized in Table 2.1.
It appeared that the organic pollutants originate from baths with greywater and
wastewater is quiet high in terms of BOD5 and COD. Furthermore, these
characteristics vary with time and geography. For instance, Teh et al., (2015) stated
that the BOD5 in greywater from bathrooms in Malaysia was 349 mg/L while a study
conducted by Al-Hamaiedeh & Bino (2010) in Jordan showed that BOD5 ranged
between 110–1240 mg/L.
A study carried out on the greywater from bathrooms by Mohamed et al., (2014) in
Malaysia showed a significant disparity of elevated COD (445-621 mg/L) compared
to the 77-240 mg/L in the study performed by Eriksson et al., (2002) on greywater
from showers and hand basins in Denmark. Furthermore, Nnaji et al., (2013a) reported
high concentrations of TSS (633.23 mg/L) in greywater from bathrooms in Nigeria.
In Greece, Fountoulakis et al., (2016) found that COD in bathrooms and laundry was
645 mg/L while PO4 was 101 mg/L which is quite high. Indeed, most of the
phosphorus from household are found in washing soap and detergent. However, the
high concentrations of nitrogen in greywater might be caused by some people who
urinate when bathing. Besides, in some areas, the kitchen and bathroom wastewater
are mixed, and this might explain the high content of phosphorus in greywater.
Mohamed et al., (2013a) revealed that the greywater in Malaysia contained 10-38
mg/L of nitrogen and 3-20 mg/L of phosphorus.
11
Tab
le 2
. 1:
Char
acte
rist
ics
of
Bat
hro
om
Gre
yw
ater
and R
elat
ed L
iter
ature
Sourc
es
Res
earc
her
s F
oun
toula
kis
et a
l.,
(20
16)
Teh
et
al.
,
(20
15
)
Mo
ham
ed
et
al.
(20
14
)
Nnaj
i et
al.
, (2
01
3)
Go
kula
n
et
al.
,
(20
13
)
Mo
ham
ed
et
al.
,
(20
13
)
Chai
llo
u e
t
al.
, (2
01
1)
Al-
Ham
aied
eh
and
B
ino
(20
10
)
Eri
kss
on
(20
09
)
Eri
kss
on
et
al.
,
(20
03
)
Chri
sto
va-
Bo
al
et
al.
,
(19
96
)
So
urc
es
Bat
hro
om
and
laund
ry
Bat
hro
om
B
athro
om
B
athro
om
M
en
ho
stel
Bat
hro
om
and
laund
ry
Bat
hro
om
V
illa
ge
ho
use
s
Bat
hro
om
S
ho
wer
s/
han
d
bas
ins
Bat
hro
om
Co
untr
y
Gre
ece
Mal
aysi
a
Mal
aysi
a
Nig
eria
In
dia
M
alaysi
a
Fra
nce
Jo
rdan
D
enm
ark
D
enm
ark
A
ust
rali
a
pH
6
.4-1
0
6.1
3
6.1
- 6
.4
7.7
7
.48
6.8
-7.8
7
.58
6.9
–7.8
N
R
7.6
-8.6
6
.4-8
.4
CO
D (
mg/L
) 2
6-6
45
44
5
44
5-6
21
67
.64
98
6
18
0-2
91
39
9
92
–22
63
NR
7
7-2
40
NR
BO
D5 (
mg/L
) N
R
34
9
40
– 1
05
60
.23
27
9.6
9
0-1
30
24
0
11
0–1
240
NR
2
6-1
30
76
-20
0
TS
S (
mg/L
) 7
-25
0
81
78
- 1
63
63
3.2
3
56
4
88
-11
0
12
5
23
–35
8
NR
7
-20
7
48
-12
0
E.c
oli
(CF
U/1
00
ml)
0-3
.4x1
05
NR
N
R
6.9
7x1
09
NR
N
R
4.7
6x1
05
NR
N
R
<1
00
-
28
00
NR
NH
4-N
(mg/L
)
NR
N
R
0.0
22
-6.7
N
R
14
.21
0.2
4-5
.2
NR
N
R
NR
0
.02
-0.4
2
<0
.1-1
5
TN
3
.6-2
1
NR
1
0-3
8
NR
1
4.2
1
5.3
-30
3.8
-17
.0
38
-61
NR
3
.6-6
.4
NR
TP
0
.1-1
01
NR
3
-20
N
R
9.6
4
0.2
2-6
.7
0.1
-2.0
N
R
NR
0
.28
-
0.7
79
NR
Cd
N
R
NR
N
R
NR
N
R
NR
N
R
0.0
08
0.6
2
NR
<
0.0
1
Pb
NR
N
R
NR
N
R
NR
N
R
NR
1
.19
3.1
-6.3
N
R
NR
Mg
N
R
NR
2
-5.2
N
R
NR
2
.1-9
.8
NR
N
R
NR
2
0.8
-23
1.4
-2.9
Cu
N
R
NR
N
R
NR
N
R
NR
N
R
NR
N
R
NR
0
.06
-0.1
2
Bo
ron
NR
N
R
0.0
9-1
.78
NR
N
R
0.0
7-0
.62
NR
N
R
NR
N
R
NR
Ca
NR
N
R
6-2
0
NR
N
R
7.5
-20
NR
N
R
NR
9
9-1
00
3.5
-7.9
Na
NR
N
R
72
-85
NR
N
R
41
-18
0
NR
N
R
NR
4
4.7
-98
.5
7.4
-18
K
NR
N
R
0.8
5-2
.5
NR
N
R
0.0
5-1
.5
NR
N
R
NR
5
.9-7
.4
1.5
-5.2
*N
R=
No
t re
po
rted
12
These differences could be related to the nature of living, geographical location,
demographics, level of occupancy, social habits and water usage patterns and time
(Jamrah & Ayyash, 2008; Prathapar et al., 2005). Besides, these comparisons were
made between the quality of potable water in developing countries and developed
countries.
2.3 Greywater Production
Greywater volume production from households is generally not measured, and
therefore needs to be estimated from alternative data sources (Hyde, 2013). Greywater
volume production for households may be estimated given the water consumption of
a household, some knowledge on how this water is used and whether water
conservation measures are being practiced in the home amongst other factors.
Antonopoulou et al., (2013) agrees that the production of greywater is directly related
to the consumption of water in a household and is dependent on a number of factors
including the level of service provision, tolerance of residents to pollution and the
community's level of awareness of health and environmental risks. They point out that
it could be assumed that greywater accounts for virtually all water usage in non-
sewered areas except for that which is used for drinking purposes, for cooking and the
water that remains on the surface of washed articles. Wood et al., (2001) noted that
there is a general absence of data on the quantification of greywater in village
settlements, owing to the fact that generally there is no proper measurement in these
areas and assumptions are made based on the population and usage of water obtained
from survey questionnaires.
During on site survey, residents indicated water consumption figures ranging from 4.7
to 28 L/c/day, although in the general absence these figures do not accurately reflect
the total amount of water drawn from the system (i.e. leaks, under-reporting are not
accounted for) (Wood et al., 2001). Furthermore, the study by Schalkwyk, (1996)
estimated the water used for dish washing, cleaning the house, clothes washing and
personal hygiene varies approximately from 12 to 50 L/c/day. Under such
circumstances Schalkwyk concluded that a greywater volume of 150 L/household/day
13
is possible, assuming a mean size of household occupants is 6, and the fact that up to
half of the water used for washing could be retained on surfaces. It is estimated that
the average person in developing countries uses 60-150 L/day (Mofokeng, 2008), and
if 90% of this ends up as greywater, then between 55 and 135 L of greywater per day
would be produced per person. He further states that for households with a standpipe
outside the house or tap in the home (not connected to a waterborne sewage system),
the amount of greywater produced is likely to be between 30 and 80 liters per day. In
places where water has to be brought in from a source at least 250 meters away from
the home, a consumption of 9-50 L/person/day is likely. Greywater volumes may then
be estimated from these figures.
However, as discussed earlier, there is high variability associated with the production
of greywater. To best estimate the amount of greywater produced by a household, it is
most accurate to determine the potential for greywater production by the household
usage habit, appliances at individual household premises and the presence of infants
and people with a wide distribution of ages (Edwin et al., 2014). The amount of
greywater produced by the households’ shower(s) can be determined by a
corresponding number of people or the type of shower head. It is assumed that in some
households, the average shower time is seven minutes (Coghlan & Higgs, 2003) and
that every resident takes a shower once daily. Thus, water used for bathing per capita
could be estimated daily. However, differences may arise between houses with piped
water supply system as water is available in close proximity of the consumers than un-
piped houses.
The volume of greywater produce from households was measured based on the number
of vessels used for each activity or is assessed by a known volume of buckets used
(Maya et al., 2016). Where running tap was utilized, the quantity of water per minute
that is coming out was multiplied and the volume is estimated (Singh & Turkiya,
2013).
From the literature review, Table 2.2 summarizes household greywater production.
The amount produced from bathroom greywater is quite different from household to
household and geography. These characteristics differ with time and geography. For
instance, Edwin et al., (2014) found 35 L/P/day from bathroom greywater compared
14
to that reported by Amin & Mahmud, (2011) with 46.6 L/p/day from household
greywater. This difference would be related to different sources of greywater and
activities of the residents from different households. A study by Shaban & Sharma,
(2007), 25.82 L/p/day was compared with a high amount of production of 52.3 L/p/day
in Edwin et al., (2014) study where both studies involved bathroom greywater. This
might be resulted from activities such as shaving, washing hair during bathing and
tooth brushing.
Table 2. 2: Quantity of Greywater Produced from Shower and Bathroom Sources
Compared with Literature Data
References Greywater
sources
Type of
household
Amount
(L/c/day)
Country
Golda et al.,
(2014)
Shower
and Bath
Urban
house
35 Puducherry
(India)
Amin &
Mahmud
(2011)
Bath and
washing
Village
house
46.6 Dhaka
(Bangladesh)
Golda et al.,
(2014)
Shower
and Bath
Urban
house
52.3 Amsterdam
(Netherland)
Shaban &
Sharma
(2007)
Shower
and Bath
City
house
25.82 India
The number, lifestyle, age, presence of children, health status and water usage patterns
of the occupants are found to affect the characteristics of greywater produced in a
household (N S W, 2007). The composition of greywater varies widely from household
to household and is highly dependent on the usage of detergents, cosmetics as well as
other personal habits of the residents. Greywater from homes with children tends to
contain higher counts of coliform than homes without children. Greywater is typically
characterized by very high concentrations of biodegradable organic material. Though
the presence of pathogenic microbes is minimal in greywater (Mara & Kramer, 2008),
it favors the growth of microbes and can turn anoxic, emanating foul odor if left
untreated for more hours. Therefore, point source treatment is considered favorable as
15
it not only enables the treatment of greywater as soon as it is produced, but also reduces
the load on the centralized treatment facility supported by the local municipalities. In
India, the goal of every state is to become sustainable through proper planning to avoid
conflicts with neighboring states in case of water dependency. Standards for greywater
were suggested by the central pollution control board (CPCB). Greywater
characteristics also vary according to its origin and for this reason; sources of
household greywater should be prioritized for treatment before discharge. The
literature data reveal that greywater from bathroom sources accounts for about 50–60
percent of total greywater (Poyyamoli et al., 2013; Coghlan & Higgs, 2003) and are
contaminated with large quantities of oils, body fats and chemicals originating from
soap, shampoo, hair dyes, toothpaste, nutrients and other cleaning products. It also
contains traces of fecal contamination (N S W, 2007). The greywater from laundry
sources accounts for about 4 % of total greywater (Coghlan & Higgs, 2003) and all
washing requirements account for about 25–30 % of total greywater (Poyyamoli et al.,
2013).
At present, there are no uniform quality standards for greywater reuse or discharge in
many developing countries including Malaysia, and the available treatment
technologies are mostly proprietary and unclear in many aspects. Also, there are no
laws or regulations on the treatment of greywater in many countries including India
(Allen et al., 2010). In Malaysia, no greywater standard or guideline exist, even though
Industrial Water Quality Standard (IWQS) does exist (Standard A and B).
2.4 Standard Regulations
The quality of wastewater is often compared with a standard guideline. Table 2.3
shows various parameters with acceptable levels of pollutants for surface water
discharged from different countries. By comparing the parameters of greywater with
Table 2.3 below, we could determine whether the greywater is contaminated and
whether it can be used as a medium for algae growth. Algae growth in greywater has
the ability to reduce the content of nutrients, thus producing parameters that are closer
to treated water. Through this study, the results of physical and chemical parameters
of greywater will be compared with the guidelines shown in Table 2.3 and Table 2.4
16
for discharge into surface water and environment quality regulations, 2009
respectively. Furthermore, the Malaysian Environment Quality (Sewerage and
Industrial Effluents) Regulations, 2009 specifies two standards for effluent discharge
to stagnant water bodies where Standard A is discharged at the upstream and Standard
B is discharged downstream. Since there is no specific standard for greywater effluent
discharge, standard A and B were used in comparison with bathroom greywater
characteristics. Table 2.4 shows the details of this standard according to the
Environment Quality Act, 1974, Regulation 8(1), 8(2) and 8(3).
Table 2. 3: Effluent Standards in Selected Countries for Discharge into Surface
Water (Alderlieste et al., 2006)
Country Costa Rica India Israel Sri Lanka Switzerland
pH 5-9 5.5-9 7-8.5 6-8.5 Na
TSS (mg/L) Na 100 10 50 Na
COD (mg/L) Na 250 70 250 Na
BOD(mg/L) 40 30 10 30 20
NH4-N (mg/L) Na 50 1.5 50 2
TN (mg/L) Na Na 10 Na Na
TP (mg/L) Na Na 0.2 Na Na
Na= Not available
Table 2. 4: Environment Quality (Sewerage and Industrial Effluents) Regulations,
2009 (Environment Quality Act 1974)
Parameter Unit Standards
A B
pH Value - 6.0-9.0 5.5-9.0
BOD5 at 20°C mg/l 10 20
Total
Phosphorus
mg/l 2 5
Nitrate-N mg/l 5 5
Ammoniacal
Nitrogen
mg/l 2 2
17
2.5 Response Surface Methodology (RSM)
In conventional multifactor experiments, optimization is generally carried out by
varying a single factor while keeping all other factors fixed at a definite set of
conditions. It is not only time-consuming, but also usually incapable of reaching the
true optimum as it ignores the interactions among variables. Alternatively, the RSM
has been proposed to determine the effects of individual factors and their interactive
influences. The RSM is a statistical method for designing experiments, building
models, evaluating the effects of several factors, and searching for optimum conditions
for desirable responses.
RSM is a capable way of achieving such an optimization by analyzing and modeling
the effects of several variables and their responses and finally optimizing the process.
This method has been widely used for the optimization of different processes in food
chemistry, material science, chemical engineering and biotechnology (Liu et al., 2014;
Singh & Sharma, 2012; Granato et al., 2010). In the traditional experimental design
approaches used in RSM such as central composite design, the number of coefficients
of the quadratic model equation increases exponentially with an increase in
experimental factors and so does the number of experimental trials (Cheng et al.,
2002).
2.6 Artificial Greywater
Artificial greywater was formulated to contain constituents typically found in actual
greywater including nutrients. To evaluate and compare the performances of several
treatment processes, experiments were conducted on artificial greywater (AGW) so
that the effluent was perfectly reproducible and representative of household greywater.
AGW has to be reconstituted so that the values of its physico-chemical and
microbiological parameters are coherent with those of real greywater. Some authors
have used a mixture of chemical substances and commercial hygiene products to create
the greywater load (Diaper et al., 2008). These products have a composition that
changes with time and most of them cannot be found all over the world; to ensure its
18
reproducibility, the AGW is composed exclusively of chemical products of technical
quality (Hourlier et al., 2010).
However, different artificial greywater recipes have been found in the literature and
they are summarized in Table 2.5. This includes a recipe that was reported
byDalahmeh et al., (2014) and Jeffrey & Jefferson, (2003). Both researchers used
shampoo, oil and laundry detergents in their artificial greywater recipe which is in
contrast with the artificial greywater recipe reported by Cardoso & Antunes, (2016)
and Diaper et al., (2008). Both researchers used almost the same recipe in their studies
(see Table 2.5).
In a study conducted by Cardoso & Antunes, (2016), artificial greywater was used to
estimate the performance of the experimental greywater treatment system. Artificial
greywater was used to assess the treatment ability of a pilot-scale MBBR. The average
dissolved oxygen concentration inside the MBBR was 7 mg/L, which is considered a
high value. Compared with real greywater, the non-biodegradable organic matter
fraction was higher in artificial greywater.
Table 2. 5: Artificial Greywater Recipe
Mariana et al.,
2016
Diaper, 2008 Jefferson et
al., 2003
Dalameh et
al., 2014
Shampoo 80.56 g 7.2 g 1.6 ml 16 g
Oil - 70 mg 80 mg 10 g
Deodorant 8.5 g 100 mg - -
Moisturizer 127.2 g 100 mg - -
Toothpaste 12.7 g 325 mg - -
Laundry
powder
150.4 ml 15 g - 16 g
Na2SO4 15 g 350 mg - -
NaHCO3 7.5 g 250 mg - -
Na2PO4 15 g 390 mg - -
Boric acid 14 mg - -
Lactic acid 12.7 g 280 mg - -
Soap 97.52 g - 620 mg -
Hair - - 20 mg -
Softener 78.9 ml - - -
Conditioner 89 g - - -
The greywater used in this experiment was artificially prepared using soap, shampoo,
shower gel, toothpaste, shaving and moisturizing cream, make-up and make-up
remover (10 g/L) as described by Saumya et al., (2015).
19
2.7 The Trend of Greywater Treatment
Greywater treatment technologies involve a combination of preliminary (physical),
primary (chemical) and secondary (biological) systems. These technologies are
emphasized due to their economic and ecological benefits, less or no skill personnel is
required, ease of handling and high treatment efficiency. In this section, the role of
each stage of the treatment system was discussed in terms of nutrient removal and
improvement of treated greywater quality.
2.7.1 The Use of Natural Filter Material in Greywater Treatment
The primary treatment for greywater mainly removes pathogens and suspended solids.
The system was found to improve turbidity and contribute to the reduction of chemical
oxygen demand (COD), biological oxygen demand (BOD), phosphorus (P), nitrogen
(N), total suspended solid (TSS), total and faecal coliforms. (Mohamed et al., 2014a).
The primary treatment mostly consists of coarse sand and soil filtration where the
coarse filter alone has limited effect on the removal of pollutants present in the
greywater. Hence it is usually combined with soil filtration and this is known as the
hybrid treatment process.
Several types of natural materials such as sand bed, fine particles, coarse sized bricks,
charcoal bed, ceramic, clamshell, limestone, wooden saw dust bed and coconut shell
covers have been combined to design a filter bed in the filtration unit (Mohamed et al.,
2014b; Karlsson et al., 2013). The utilization of natural materials as filtration units
such as constructed wetlands have exhibited high efficiency in removing pollutants. It
is also inexpensive and involves simple operations (Siracusa & La Rosa, 2006).
Mohamed et al., (2014a) found that the filtration system consisting of peat, charcoal
and gravel was effective for the treatment of greywater. The filtration system which
consisted of bark and activated charcoal also exhibited efficiency in reducing BOD5
and total phosphorus (TP) by more than 90% (Karlsson et al., 2013).
Table 2.6 compares different low cost treatment schemes on greywater from different
sources including several households (Gross et al., 2007), mosques (Mohamed & Ali,
20
2012), residential quarters (Nnaji et al., 2013), village houses and house kitchens
(Mohamed et al., 2013). The main components which are removed by using the
primary treatment system are BOD, COD, TSS with efficiency ranging from 37 to
98%, 74 to 90.8% and 40 to 95% respectively (Dalahmeh et al., 2012; Gross et al.,
2007). In a review by Al-Jayyousi, (2003), a simple greywater treatment system
consisting of a sand filtration unit reduced SS and BOD by 40 and 74%.
21
Table 2. 6: Review of Primary Treatment System of Different Sources of Greywater
Location Greywater Treatment
System
Greywater
Sources Pollutant Removal References
Nsukka,
Nigeria.
Gravity System by
sedimentation unit,
sand & gravel filtration
unit, receiving unit,
adsorption unit, storage
unit
Resident
quarters
BOD (85.68%)
COD (57.09%)
TSS (70.74%)
FC (100%)
Nnaji et al.,
(2013)
Madaba
Governorate,
Jordan
Filter Media:
Wetland bed
Volcanic Tuff
(volcanic ash)
White gravel
Al-Faisalia,
Al-Areash and
Jraineh village
houses
BOD (73%)
COD (65%)
TSS (84%)
FC (15.67%)
Mohammed
et al., (2013)
Cairo, Egypt
Combine Chemical
System by:
Coarse &
surge tank
Sand filter
with reeds
Mosque,
shower
bathroom of
housing
building and
sinks of the
bathroom
BOD (71%)
COD (67%)
SS (87%)
Turbidity (90%)
TC (100%)
Mohamed et
al., (2012)
United State
Vertical flow
constructed wetland
used:
Peat Moss
Lime pebbles
Several
households
COD (90%)
TSS (95%)
E. coli (100%)
Gross et al.,
(2006)
Tafileh, Jordan
Separation and
Automatic greywater
system (automatic back
washing sand filter)
Houses and
institutional
buildings
TSS (40%)
BOD (74%)
Al-Jayyousi
et al., (2003)
Swedin
Pine bark
Activated
carbon
Foam
Sand filters
Laboratory
(column
experiment)
BOD, COD, TN, TP
(98, 74, 19 and 97%
respectively)
BOD, COD, TN, TP
(97, 94, 98 and 91%
respectively)
BOD, COD, TN, TP
(37, 37, 13 and 36%
respectively)
BOD, COD, TN, TP
(75, 72, 5 and 78%
respectively)
Sahar et al.,
(2012)
Malaysia
Gravel, Peat, Charcoal
and sand filter media
House Kitchen
Wastewater
BOD5 40%, CODtot
37%,SS 72%, NH+4-N
87% and pH 6.6-6.7
Mohamed et
al., (2013a
and b)
Australia
Land and Water
Greywater system with
dual sponge filter
House
(laundry &
bathroom)
BOD 24-200 mg/L,
COD 35-739 mg/L, TSS
78.33-163 mg/L, TP 3-
20 mg/L
22
Greywater from kitchens contains much higher concentrations of organic substance,
nitrogen, oil and grease and detergents from the dishwashing process. Mohamed et al.,
(2013) showed that the efficiency of a treatment system consisting of gravel, sand, peat
and charcoal as treatment media to remove nutrients and organic compunds in kitchen
greywater was 72% for SS, 37% for CODtot, 40% for BOD5, and 87% for NH4+-N.
This signifies that peat soil can also be a potential material for removing pollutants.
Hence, kitchen greywater can be treated with peat media. Furthermore, according to
Table 2.6, total nitrogen removal varies from 5-98% while phosphorus removal ranged
from 36-99.9%.
Kariuki et al., (2011) investigated a 5-barrel greywater treatment (GWT) system
consisting of 5 recycled polyethylene (PE) plastics barrels linked by polyvinyl chloride
(PVC) pipes. The treatment system was connected with an ordinary sieve as a pre-
treatment of greywater. The rock alum (aluminium sulphate) was added to the
greywater then disinfected with sodium hypochlorite before reuse. The GWT system
generates effluent which meets the guidelines by the World Health Organization
(WHO) to be reused for irrigation. The designed GWT system was found to reduce
faecal coliforms as well as Salmonella sp. in greywater before being used for
subsurface irrigation.
Dalahmeh et al., (2012) studied the efficiency of activated charcoal, pine bark, sand
filters and polyurethane foam for the reduction of contaminants from artificial
greywater in a laboratory column experiment. Pine bark and activated charcoal filters
showed high efficiency for reducing BOD5 by 98 and 97% and methylene blue active
substances (MBAS) by 99 and 99% as well as TP by 97 and 91% respectively. The
efficiency recorded in that study might be related to the absorption process associated
with the presence of a high percentage of sodium chloride (NaCl) and uranin (93%)
traces in the pine bark filter. The study indicated that activated charcoal and pine bark
might be effective for producing treated greywater suitable for irrigation. Besides, the
study demonstrated that the pine bark filters might be more useful for the effluent
which would be used for irrigation, as it preserves and restores the use of chemical
fertilizer due to its high concentration of nitrogen.
23
Nnaji et al., (2013) reported the sand filter treatment method. The study reveals a
significant removal of organic compounds, physical and microbial pollutants such as
BOD5 (85.68%), COD (57.09%), TSS (70.74%) and faecal coliform by 99.99%
respectively. The chemical composition of steel slag, clamshell and limestone used in
the natural filtration system exhibited efficiency for the removal of pollutants from
greywater. Clam shell and limestone contain an alkaline agent (CaO) which plays an
important role in neutralizing or partially neutralizing acidity as well as the adsorption
process of suspended solids. Steel slag that contains CaO, Fe2O3 and SiO2 with
different percentages increase the removal efficiency of TSS (Bird & Drizo, 2010).
This filtration might also contribute to the reduction of total phosphorus and nitrogen
during the filtration of wastewater and might be used to improve the quality of
secondary effluents (Zhang et al., 2015).
Clamshells are made of aragonite, a form of calcium carbonate (CaCO3). Thus, it has
the potential to remove BOD5 efficiently. In addition to removal, sand filtration also
removes the BOD5 contained in the suspended particles (Libhaber & Jaramillo, 2012).
In a previous study performed by (Park, 2009) and (Luo et al., 2013), the treatment
system using oyster shells removed BOD5 by 89.3% and 85.02% respectively. It was
because oyster shells are rich in calcium oxide which exhibits high biosorption
efficiency. In the present study, the filtration system was multilayered to increase the
removal efficiency of COD and BOD5. (Liu et al., 2010) also reported that the two-
layer filter was more efficient for the removal of BOD5 than the single layer filter (85
vs. 76%). Liu et al., (2010) showed that the filtration system of wastewater using oyster
shell reduced COD by 85.1%.The application of sand and clamshell filtration reduced
the turbidity by 90 and 89.9% respectively (Mohamed et al., 2014; Park, 2009).
Niwagaba et al., (2014), investigated the treatment of greywater via a multi-media
filter consisting of gravel, charcoal and geotextile. It was noted that the filtration
system reduced TSS by 85.2%. The reductions in COD and BOD5 were 90.8 and
96.1%, respectively. Karlsson et al., (2013) found that the filtration system consisting
of bark and activated charcoal removed more than 90% of BOD5. In contrast,
Mohamed et al., (2014a) studied the efficiency of a natural filtration system consisting
of a two-stage filter media, pre-treatment (gravel and sand) and peat based (peat,
24
charcoal and gravel) for the treatment of household greywater. However, the reduction
of TSS, BOD5 and COD was 81%, 54%, and 52% indicating that gravel and charcoal
are not the main factors which affect the efficiency of filtration system.
The study carried out by Mohamed et al., (2013) on the natural filtration system of
greywater indicated that the quality of the treated wastewater complied with the limits
of the Malaysian Standard (Standard B). The treated greywater using septic tank
followed by intermittent sand filter in Jordan has met the Jordanian Standards JS
(893/2006) for the Reclaimed Wastewater reuse for restricted irrigation (Assayed et
al., 2010). However, EQA 1974 has higher standards and the wastewater did not meet
the EQA 1974 standards A. Moreover, using a treatment system consisting of
biological material might produce high quality greywater.
2.8 Potential Use of Microalgae in Treating Greywater
Algae are simple plants that can range from microscopic (microalgae), to large
(macroalgae) photosynthetic organisms. Microalgae includes both cyanobacteria
(blue-green algae) as well as green, brown and red algae which are predominantly
multicellular although some types are unicellular organisms. These organisms are
primary producers of organic matter in aquatic environments. Inorganic compounds
like ammonia, nitrate, phosphate and carbon dioxide provide food source to synthesize
new algal cells and produce oxygen. In the absence of sunlight, algae use energy
released by inorganic chemical reactions to produce food through daily variation in
DO levels and consume oxygen in water.
Microalgae are prokaryotic or eukaryotic photoautotrophic microorganisms that grows
in all kinds of environment due to their simple morphological structure. Prokaryotic
microalgae are identified as cyanobacteria (Cyanophyceae) and eukaryotics includes
green algae (Chlorophyta) among other groups (Mata et al., 2010). Green algae are the
most diverse group of algae growing in a variety of habitats and are considered
paraphyletic because it excludes plantae. The green algae contains two forms of
chlorophyll (chlorophyll a & b and carotenoid), which they use to capture light to fuel
the manufacture of sugars (Parlevliet & Moheimani, 2014; Singh & Sharma, 2012).
113
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