WATERING OF LANDSCAPE PLANTS USING WASTEWATER TREATED
WITH TERMINALIA CATAPPA
NURFARHAIN BINTI MOHAMED RUSLI
A report submitted in partial fulfillment of the
requirements for the award of the degree of
Master of Engineering
Faculty of Civil Engineering
Universiti Teknologi Malaysia
JUNE 2009
iii
“To my loves family…..”
Mama, Ablong, Kak Aitul, Abngah, Jiejah,
Farah and Fathin…
Thanks for your support,
To my loves one, thanks for your love, care
and encouragement….
All friends…..
Thank you for everything…..
Our Loves Never Ends
iv
ACKNOWLEDGEMENT
“In the name of God, the most gracious, the most compassionate”
First of all, thank god for giving me the strength to complete this thesis within
the time.
Then, I would like to express my deep appreciation to my beloved family
members:, mother, and my siblings who always providing me the encouragement and
spiritual support to choose the way of life which I believe on.
My thesis supervisor, Dr Shamila Azman is an important figure in guiding and
counseling me towards the thesis accomplishment. Her generosity, patience,
tolerance and helping character nourished me more than what I gained through the
successful completion of thesis. Thanks a lot, Dr Shamila!
I am also very appreciation to all environmental lab personnel, Pak Usop, En
Ramli, En Azreen, Kak Ros who have contributed their assistance and views at
various occasions.
Last but not least, I would like to convey my special thanks to all friends, and
those whoever lending a helping hand either directly or indirectly, that led to the
completion of this thesis. Thank You!
v
ABSTRACT
This study was conducted to identify the antibacterial properties of
Terminalia cattapa (TC) leaves and also to explore the feasibility of using it with
domestic wastewater effluent for reuse purposes. Previous study conducted in UTM
have shown the potential of using domestic wastewater in watering landscape plants
where chlorine was use to disinfect the wastewater. Since chlorine application
produce residual effect, TC leaves offers another potential to be applied together with
the domestic wastewater in order to eliminate bacterial contamination. From analysis
conducted, TC leaves in blended form with wastewater yield better results compared
to cut and original leave form. In this study, effluent from domestic wastewater
oxidation pond was collected and applied daily to two landscape gardens with a
surface area of 1.5 m x 1.0 m x 0.3 m where both plots were planted with five
different plants. The difference between the two plots are one is applied with
wastewater and TC leaves meanwhile the other one is applied with wastewater only
and stand as a control. Wastewater effluents collected were analysed for BOD, COD,
E. coli, phosphorus, ammoniacal nitrogen and pH. From the result obtained all
samples treated using TC leaves shows 98% removal of E. coli numbers after 5 days
treatment. In order to compare bacterial removal, analysis on residual chlorine was
also carried out, result obtained shows that even though chlorine is effective in
eliminating bacteria it has residue effect which is harmful to human or plants and
insects. Treated sewage effluent applied to landscape plants produce better growth
when compared to untreated wastewater. The results obtained showed positive
growth increment for wastewater with TC leaves in terms of height and width for all
plants except for Elephantopus scaber (Es) which thrive more when applied with
wastewater.
vi
ABSTRAK
Kajian ini dijalankan adalah untuk mengenalpasti sifat anti-bakteria dan juga
untuk mengkaji kebolehgunaan daun ketapang dalam merawat air sisa domestik
untuk tujuan guna semula. Hasil kajian lepas yang dijalankan di UTM, menunjukkan
penggunaan air sisa domestik berpontensi digunakan semula untuk menyiram
tumbuhan landskap. Dalam rawatan tersebut, klorin telah digunakan untuk merawat
air sisa tersebut serta bertindak sebagai agen anti-bakteria. Oleh kerana penggunaan
klorin meninggalkan sisa, TC menunjukkan potensi untuk digunakan dalam merawat
air sisa domestik. Daripada analisis yang dijalankan, daun ketapang yang dikisar
dengan air sisa menunjukkan keputusan jauh lebih baik berbanding dengan daun
yang dipotong atau dalam keadaan biasa. Dalam kajian ini, efluen dari kolam
pengoksidaan telah digunakan dua kali sehari pada dua taman landskap yang berbeza
dengan lima jenis tumbuhan di tanam di atas landskap bersaiz 1.5 m x 1.0 m x 0.3 m.
Perbezaan antara plot tanaman tersebut ialah satu akan disiram air sisa domestik
yang di rawat dengan daun ketapang dan yang satu lagi bertindak sebagai kawalan
dan disiram dengan air sisa domestik sahaja. Air sisa domestik yang diambil telah di
analisis untuk parameter BOD, COD, E. coli, fosforus, ammonia nitrogen dan pH.
Dari keputusan yang diperolehi, semua sampel yang dirawat dengan daun ketapang
menunjukkan penurunan E. coli sebanyak 98% selepas 5 hari. Dalam perbandingan
penyingkiran bakteria, analisis terhadap sisa klorin turut di jalankan dan hasil
menunjukkan walaupun klorin berkesan untuk membunuh bakteria, ia meninggalkan
kesan sisa yang boleh membahayakan manusia, tumbuhan dan juga serangga. Air
sisa domestik yang dirawat dan diaplikasikan ke tanaman landskap menghasilkan
pertumbuhan baik berbanding dengan air sisa yang tidak dirawat. Keputusan
menunjukkan pertumbuhan positif berlaku dalam pertumbuhan tinggi dan lebar daun
untuk semua tanaman kecuali pada elephantopus scaber (Es), yang tumbuh subur
dengan air sisa domestik yang tidak di rawat.
vii
TABLE OF CONTENTS
CHAPTER
TITLE
DECLARATION
ii
DEDICATION
iii
ACKNOWLEDGEMENTS
iv
ABSTRACT
v
ABSTRAK
vi
TABLE OF CONTENTS
vii
LIST OF TABLES
xii
LIST OF FIGURES
xiv
LIST OF ABREVATIONS
LIST OF APPENDICES
1
2
PAGE
xviii
xix
INTRODUCTION
1
1.1
Introduction
1
1.2
Problem Statement
3
1.3
Objective of the Study
5
1.4
Scope of the Study
6
1.5
Significance of Study
6
LITERATURE REVIEW
8
2.1
Introduction
8
2.2
Domestic Wastewater
9
viii
2.3
Characteristics of Domestic Wastewater
14
2.3.1 Physical Characteristics
15
2.3.2
Chemical Characteristics
18
2.3.3
Biological Characteristics
23
2.3.4 Nutrients
25
2.4
Oxidation Pond (OP)
28
2.5
Wastewater Reuse Applications
30
2.6
Potential Risks from Using Recycled Water
33
2.7
Agricultural Reuse
33
2.8
Sustainable Environment and Development
36
2.9
Global Warming and Climate Change
38
2.10
Carbon Footprint
41
2.11
Water Reuse Problem
43
2.12
Wastewater Treatment
44
2.13
Wastewater Treatment in Malaysia
45
2.13.1 Environmental Regulations and Standards in
45
Malaysia
2.14
Wastewater Treatment Method
46
2.14.1 Aluminium Sulphate (Alum)
47
2.14.2 Ferric Chloride
48
2.14.3 Ozonation
48
2.14.4 Ultraviolet (UV)
49
2.14.5 Disinfection
50
2.15
Previous Study
51
2.16
Terminalia catappa
52
2.16.1 Characteristics of Terminalia catappa
53
2.16.2 The Uses of Terminalia catappa
55
2.16.3 Tannin
58
2.17
Escherichia coli
59
2.18
NPK Ratio
60
2.18.1 The Secondary Macronutrients
62
ix
3
METHODOLOGY OF RESEARCH
64
3.1
Introduction
64
3.2
Jar Test
67
3.3
Leaf Condition Analysis
67
3.3.1
68
Optimum Dosage of Terminalia catappa Leaves
(Tannin Concentration)
3.4
Extraction of Terminalia catappa Leaves
69
3.5
Wastewater Collection
70
3.6
Sample Handling Procedure
71
3.7
Apparatus and Chemicals
71
3.8
Wastewater Treatment using Terminalia catappa Leaves
73
3.9
Water Quality Analysis
73
3.9.1 pH
74
3.9.2
75
3.10
Biochemical Oxygen Demand (BOD)
3.9.3 Chemical Oxygen Demand (COD)
75
3.9.4 Ammoniacal Nitrogen (AN)
76
3.9.5 Phosphorus
77
3.9.6 Tannin
77
3.9.7
Escherichia coli
77
Disinfection of Effluent
79
3.10.1 Iodometric Method for Determinataion of
80
Residual Chlorine
4
3.11
Landscape Design
81
3.12
Plants Growth Determination
83
3.13
Preparation of Agar Nutrient (EMB Agar)
84
3.13.1 Plate Colony Counting
86
3.13.2 Serial Dilutions for Plate Counting
87
3.14
Microbiology Sampling Procedures
89
3.15
Determination of NPK Ratio
90
RESULT AND ANALYSIS
91
4.1
91
Introduction
x
4.2
4.3
Optimum Dosage of Terminalia catappa (TC) Leaves
91
4.2.1
92
Analysis of Tannin Concentration
4.2.2. Analysis of pH
93
4.2.3 Soxhlet Extraction
95
Wastewater Treatment Analysis
96
4.3.1 pH
96
4.3.2
97
Biochemical Oxygen Demand (BOD)
4.3.3 Chemical Oxygen Demand (COD)
4.4
99
4.3.4 Ammoniacal Nitrogen
100
4.3.5 Phosphorus
101
4.3.6
102
Escherichia coli
Analysis of Coliform on Leaves and Grass
103
4.4.1 Coleus artopurpureus (Ca)
104
4.4.2
Cyperus dubius rottb (Cd)
105
4.4.3
Nyctanthes arbo-tristis L (Na)
106
4.4.4 Panicaum maximum cv. colonio (Pm)
107
4.4.5
Vinca rosea (Vr)
109
4.4.6
Elephantopus scaber (Es)
110
4.5
Analysis of Coliform on Soil from landscape plots
111
4.6
Wastewater Influence on Plant Growth
112
4.7
Growth Rate of Plants
116
4.7.1
Growth Rate of Vinca rosea (Vr)
116
4.7.2
Growth Rate of Coleus artopurpureus (Ca)
118
4.7.3
Growth Rate of Nyctanthes arbo-tristis L (Na)
119
4.7.4
Growth Rate of Panicaum maximum cv. colonio
121
(Pm)
5
4.7.5
Growth Rate of Cyperus dubius rottb (Cd)
122
4.76
Growth rate of Elephantopus scaber (Es)
124
4.8
Nitrogen-Phosphorus-Potassium Ratio (NPK)
125
4.9
Disinfection
126
CONCLUSION AND RECCOMMENDATION
129
5.1
129
Conclusion
xi
5.2
Recommendation
131
REFERENCES
133
APPENDIXES
144
xii
LIST OF TABLES
TABLE NO.
2.1
TITLE
PAGE
Parameter Limits of Effluent of Standard A and B
11
(Environment Quality Act, 1974)
2.2
Composition of Typical Untreated Domestic Wastewater
15
(Burks and Minnis, 2006)
2.3
Typical Composition of Untreated Domestic Wastewater
21
(Metcalf and Eddy, 2004)
2.4
Typical of Organisms of Untreated Domestic Wastewater
26
(Metcalf and Eddy, 2007)
2.5
Typical and Number of Microorganism of Untreated
27
domestic (Metcalf and Eddy, 2007)
2.6
Effluents of Oxidation Pond (Indah Water Consortium, 2008)
29
2.7
Advantages and Disadvantages of Wastewater Reuse
31
(Kumar et al., 2006)
2.8
Standards of Treated Wastewater for Irrigation in India
35
(Kumar et al., 2006)
3.1
List of Apparatus and Chemicals for each Ttest
72
4.1
Concentration of Tannin and pH for Terminalia catappa Leaves
95
extracted with Soxhlet Apparatus
4.2
Average growth rate of plants from experiment and control
113
landscape garden
4.3
pH values for different soil samples collected from landscape
115
garden
4.4
NPK ratio of different soil in landscape garden
125
xiii
4.5
Effect of chlorine dosage and contact time towards E. coli
126
survival rate
4.6
Total residue of chlorination using Iodometric Method
128
xiv
LIST OF FIGURES
FIGURE NO.
TITLE
PAGE
1.1
Percentage of Activity Contribute to Water Pollution (DOE, 2005) 4
2.1
The increasing concentration of greenhouse gases from the year 0 39
to 2005 (IPCC, 2007)
2.2
The greenhouse effect is where most infrared radiation is
40
absorbed by the atmosphere and re-emitted in all directions by
greenhouse gas (GHG) molecules and clouds (IPCC, 2007)
2.3
Terminalia catappa tree
53
2.4
Illustration of Terminalia catappa Leaves
53
2.5
Terminalia catappa leaves Before Falling
54
2.6
Terminalia catappa leaves After Falling
54
2.7
Terminalia catappa Flowers
54
2.8
Terminalia catappa Fruits
54
2.9
Escherichia coli (E. coli) (Gerard et al., 2004)
60
3.1
Design Framework of Part One for this study
65
3.2
Design Framework of Part One for this study
66
3.3
Jar Test Analysis
67
3.4
Jar Test analysis for optimum dosage of tannin concentration
68
3.5
Example of Soxhlet Extraction carried out at Organic Research
69
Laboratory, Faculty of Science (UTM)
3.6
Example of Soxhlet Extractor with Terminalia catappa Leaves
70
3.7
Oxidation pond near Kolej Tunku Canselor, UTM
71
3.8
Treatment of wastewater using Terminalia catappa Leaves in the 73
tank
xv
3.9
Meter pH model Orion 420 A
74
3.10
BODTrak Apparatus
75
3.11
COD Reactor HACH DRB200
76
3.12
HACH DR5000 Spectrophotometer
77
3.13
Analysis of E. coli using Quanty-Tray/2000
78
3.14 (a)
Quanti-Tray Sealer Model 2x
78
3.14 (b)
Thermolyne Type 142300 Incubator
78
3.15
Chlorination process using magnetic stirrer (Chemix model CL 6) 79
3.16
Landscape Garden Side View
81
3.17
Landscape Garden Plan View
82
3.18 (a)
Landscape Garden with Coleus artopurpureus (Ca)
82
3.18 (b)
Landscape Garden with Nyctanthes arbo-tristis L (Na)
82
3.19 (a)
Landscape Garden with Five Different types of plants
83
3.19 (b)
Landscape Garden with Vinca rosea (Vr)
83
3.20
Width Growth Determination
84
3.21
Height Growth Determination
84
3.22 (a)
Figure of preparation of agar nutrient
86
3.22 (b)
Figure of agar was poured in petri dishes
86
3.23 (a)
Figure indicates that the agar nutrient in petri dish (before)
87
3.23 (b)
Figure indicates that the agar nutrient in petri dish (after)
87
3.24
Plate Counts and Serial Dilutions (Gerard et al., 2004)
88
3.25 (a)
Figure shows that cotton buds that were used to swab the leaves
89
3.25 (b)
Figure shows that (Bunsen burner that will used to heat up the
89
cotton bud before use
4.1
Concentration of tannin from Terminalia catappa leaves in
92
different forms i.e. original leaf, cut and blended in distilled water
4.2
pH measurement of TC leaves from 30 g to 70 g of
94
immersion in distilled water after 7 days
4.3
pH measurement for distilled water added with 70 g TC leaves
94
in original, cut and blended form
4.4
pH of domestic wastewater before and after treatment with
97
TC leaves for 5 days
4.5
Concentration of BOD in wastewater samples before and after
98
xvi
treatment with TC leaves for 5 days
Concentration of COD in wastewater samples collected on 2nd
4.6
99
May to 2nd Jun 2009 before and after treatment with TC leaves
4.7
:
Concentration of ammoniacal nitrogen in wastewater samples
nd
100
nd
collected on 2 May to 2 Jun 2009 before and after treatment with
TC leaves for 5 days
4.8
Concentration of phosphorus in wastewater samples collected on 101
2ndMay to 2nd Jun 2009 before and after treatment with TC leaves
4.9
E. coli numbers in domestic wastewater effluent before and after 103
treated with TC leaves
4.10
E. coli on Coleus artopurpureus (Ca) plant during different
104
sampling time intervals
4.11
E. coli on Cyperus dubius rottb (Cd) plant during different
105
sampling time intervals
4.12 (a)
Cyperus dubius rottb (Cd) plant
106
4.12 (b)
Coleus artopurpureus (Ca) with different surface leaves area
106
4.13
E. coli on Nyctanthes arbo-tristis L (Na) plant during different
107
sampling time intervals
4.14
E. coli on Panicaum maximum cv. colonio (Pm) plant during
108
different sampling time intervals
4.15
E .coli on Vinca rosea (Vr) plant during different sampling
109
time intervals
4.16
E. coli on Elephantopus scaber (Es) plant during different
110
sampling time intervals
4.17
E. coli in soil samples analysed on 11th May to 2nd June 2009
112
4.18
Flowers watered with wastewater and TC is more radiant and
114
shining compared to the control plants
4.19
Growth rate of Vinca rosea (Vr) plant in terms of height for
116
experimental plot and control plot garden
4.20
Growth rate of Vinca rosea (Vr) plant in terms of width
116
(leaves) for experimental plot and control plot garden
4.21
Growth rate of Coleus artopurpureus (Ca) plant in terms
117
of height for experimental plot and control plot garden
4.22
Growth rate of Coleus artopurpureus (Ca) plant in terms
118
xvii
of width (leaves) for experimental plot and control plot garden
4.23
Growth rate of Nyctanthes arbo-tristis L (Na) plant in terms
119
of height for experimental plot and control plot garden
4.24
Growth rate of Nyctanthes arbo-tristis L (Na) plant in terms
119
of width (leaves) for experimental plot and control plot garden
4.25
Growth rate of Panicaum maximum cv. colonio (Pm) plant in
121
terms of height for experimental plot and control plot garden
4.26
Growth rate of Panicaum maximum cv. colonio (Pm) plant in
121
terms of width (leaves) for experimental plot and control plot garden
4.27
Cyperus dubius rottb (Cd) plant with small leaves surface area
122
4.28
Growth rate of Cyperus dubius rottb (Cd) plant in terms of
122
height for experimental plot and control plot garden
4.29
Growth rate of Elephantopus scaber (Es) plant in terms of
123
width for experimental plot and control plot garden
4.30
Bacteria removal after disinfection process with
126
different chlorine dosage and contact times
4.31
Chlorine residue after disinfection process with different
chlorine dosage
127
xviii
LIST OF ABREVATIONS
o
C
degree celcius
BOD
biochemical oxygen demand
COD
chemical oxygen demand
DO
dissolved oxygen
DOE
Department of Environment
GHG
greenhouse gases
GWP
global warming potential
IPCC
Intergovernmental Panel on Climate Changes
m
meter
mg/L
miligram per litre
OP
oxidation pond
SS
suspended solids
TC
Terminalia catappa
UTM
Universiti Teknologi Malaysia
xix
LIST OF APPENDICES
APPENDIX
A
TITLE
Determination of Biochemical Oxygen Demand
PAGE
144
(BOD) Using HACH DR5000 Spectrophotometer
B
Determination of Chemical Oxygen Demand (COD)
151
Using HACH DR5000 Spectrophotometer
C
Determination of Ammoniacal Nitrogen Using
156
HACH DR5000 Spectrophotometer
D
Determination of Phosphorus Using HACH DR5000
159
Spectrophotometer
E
Determination of Tannin Using HACH DR5000
162
Spectrophotometer
F
Determination of E. coli
165
CHAPTER 1
INTRODUCTION
1.1
Introduction
Water is a common substance that is essential for the survival of all known
forms of life. In typical usage, water refers only to its liquid form or state, but the
substance also has a solid state, ice, and a gaseous state, water vapor or steam
(Metcalf and Eddy, 2004). Water covers 71% of the Earth's surface, it is found
mostly in oceans and other large water bodies, with 1.6% of water underground in
aquifers and 0.001% in the air as vapor, clouds (formed of solid and liquid water
particles suspended in air), and precipitation (Marks, 2001). Water is a tasteless,
odorless liquid at standard temperature and pressure. The color of water and ice is,
intrinsically, a very light blue hue, although water appears colorless in small
quantities. Ice also appears colorless, and water vapor is clearly invisible as a gas
(Environmental Protection Agency, 2006).
According to Malaysia Water Industry Guide (2007), the increased demand
for clean water has led to competition in water use among the various water user
sectors and the continued economic growth will magnify this even more acutely. The
2
practicable limit of surface water resources development has been reached in regions
of high demand, and it has become necessary to consider interbasin and interstate
water transfers. The current approaches towards water supply in cities are supply
driven when there’s a “shortage”, new sources are developed.
Fresh water is vital to sustain human life. However, only 3% of total water
on earth is fresh water and two-thirds of that is in frozen forms such as the polar ice
caps, glaciers and icebergs. The remaining 1% of the total fresh water is either
surface water or groundwater where groundwater consists of two-thirds of this
amount. The United States annually receives rainfall in a quantity sufficient to cover
the entire country to a depth of 30 inches that is known as the U.S. water budget
(Environmental Protection Agency, 2006). However, the annual precipitation is not
evenly distributed around the country. The eastern half of the country receives more
precipitation than the western half.
Water is the earth’s most ubiquitous and most effective dissolving agent,
playing a key role in human civilization. It quenches the thirst and enables the
growth of food and fibre for 6.1 billion human inhabitants. Humans now use half of
the readily available freshwater, which is in short supply, less than 1% of earth’s
water resources goes for domestic, agricultural, horticultural, and industrial needs.
The world population in 2008 was estimated at 7.7 billion with an annual
growth rate of 3.5 percent or 83 million people per year. To put the recent growth in
perspective, the world population in the year 1900 was only 1.6 billion and in 1950 it
was 2.5 billion. It is projected that the world population in 2050 will be between 7.9
billion and 10.3 billion (United Nations, 2008). The rate of growth in industrialized
countries is well under one percent per year. In developing countries, the growth rate
exceeds six percent per year, and in some parts of Malaysia, China andSingapore
which the growth rate increase 6.3 percent in 2008 for Malaysia. As a result, over
90 percent of all future population increses will occur in the developing world
(United Nations, 2008).
3
1.2
Problem Statement
The water demands are increasing due to the increasing population and the
rapid development in our country (Blumenthal et al., 2000).
Devamany a/l S.
Krishnasamy (Utusan Malaysia 30 Mac 2005), reported that 12.5-14 billion meter
cubic water is used by the humans in the world every year and the usage for a year
are 9000 meter cubic per capita global. Increase in population, rapid industrialization
and agricultural activities have increased the water demand to a greater extent.
In 2006, the Department of Environment (DOE) registered 18,956 water
pollution point sources comprising mainly sewage treatment plants (9,060: 47.79%
inclusive of 601 Network Pump Stations), manufacturing industries (8,543: 45.07%),
animal farms (869: 4.58%) and agro-based industries (484: 2.55%) represents the
distribution of industrial water pollution sources from agro-based and manufacturing
industries compiled by the DOE in 2006 through field surveys and questionnaires. A
total of 9,027 sources were identified with Selangor having the highest number of
water pollution sources (1,850: 20.49%), followed by Johor (1,774: 19.65%).
According to statistics compiled by the Veterinary Department of Malaysia,
the total standing pig population for 2006 was about 1.67 million, a decrease of 1.76
percent compared to 1.7 million in 2005. Correspondingly, the number of pig farms
decreased to 869 farms compared to 898 in the previous year. The number of
sewage treatment plants under the management of Indah Water Consortium Sdn.
Bhd. (IWK) had increased to 9,060 in 2006 compared to 8,782 plants in 2005.
Selangor had the largest number of sewage treatment plants (2,563: 28.3%), Perak
(1,343: 14.8%), Johor (1,010: 11.1%) and Negeri Sembilan (928: 10.2%)
Although the effluents from the sewerage treatment plant are treated, the
effluents affect the water pollution in our country particularly when the effluents are
4
discharged to the river, lake or stream (DOE, 2008). High nutrient concentration in
the effluent can contribute to water pollution which can destroy the aquatic life and
environment. Therefore, nowadays there are a lot of water pollution issues
everywhere around the world.
Water pollution occurs when a body of water is adversely affected due to the
addition of large amounts of materials to the water (Metcalf and Eddy, 2004). When
it is unfit for its intended use, water is considered polluted. Figure 1.1 below shows
the percentage of the activity that contribute to water pollution, and the higher
percentages are from sewerage treatment plant compared to other activity.
Therefore, an alternative water conversation method needs to be carry out to prevent
the problem.
Sew erage
Treatm ent Plant;
52.6%
Industry; 37.9%
Agricultural
Industry; 3.5%
Pigery livestock;
6%
Figure 1.1: Percentage of Activity Contributes to Water Pollution (DOE, 2005)
The best alternative that could be practice is reuse of the effluent in our daily
activities. In the recent year, human are more concern and aware of the serious ness
on reuse and reclamation water in the world. In many locations where the available
supply of fresh water has become inadequate to meet water needs, it is clear that the
once-used water collected from communities and municipalities must be viewed not
5
as a waste to be disposed of but as a resource that must be reused. The concept of
reuse is becoming accepted more widely as other parts of the world which is
experiencing water shortage (Metcalf and Eddy, 2004).
The use of dual water systems, such as now used in St. Petersburg in Florida
and Rancho Viejo in California, is expected to increase in the future (Metcalf and
Eddy, 2004). Most of the reuse of wastewater occurs in the arid and semiarid
western and southwestern states of the United States. However, the number of reuse
projects is increasing in the south especially in Florida and South Carolina (Metcalf
& Eddy, 2004). In Malaysia, recycled wastewater is also used for horticultural
irrigation, residential garden irrigation and toilet flushing. Typically wastewater is
produced in a larger quantity in the cities but used for agriculture in the countryside.
Malaysia faces a particularly pressing water resources challenge. It has a largely
agricultural population heavily reliant on the over exploitation of groundwater for its
survival.
1.2
Objective of the Study
The objectives of this study are:-
(i)
To identify the antibacterial properties of Terminalia catappa leaves
(ii)
To compare the usage of Terminalia catappa leaves with chlorination
process to kill bacteria in domestic effluent
(iii)
To identify the feasibility of reusing domestic effluent treated with
Terminalia catappa to water landscape plants.
6
1.4
Scope of the Study
Wastewater samples were collected from oxidation pond at Universiti
Teknologi Malaysia (UTM) Skudai. The plant is a major treatment facility in UTM,
which processed an average 250 000 m3/day of wastewater originating from campus
and hostels area. The study explores the feasibility of using effluent from domestic
wastewater treatment plant to waters landscape plants. Thus, the study will aim at
obtaining and analyzing the coliform bacteria data in order to investigate the fecal
contamination occurring on plants in the landscape garden.
Terminalia catappa leaf was used as disinfection agent to kill bacteria to
ensure public also health.
The feasibility of the wastewater reuse practice in
Malaysia was also evaluated by comparing the data with other regulations. An openair landscape garden of 1.5 m x 1.0 m x 0.3 m was designed and constructed with
five different shrubs was planted covering the area.
The plants are Coleus
artopurpureus (Ca), Cyperus dubius Rottb (Cd), Nyctanthes arbo-tristis L (Na),
Panicaum maximum cv. Colonio (Pm) and Vinca rosea (Vr).
Grass type
Elephantopus Scaber (Es) was also planted in the middle region of the garden.
1.5
Significance of Study
Chlorination is by far the most common method of wastewater disinfection
and is used worldwide for the disinfection of pathogens before being discharge into
receiving streams, rivers or oceans. Although chlorine has always been used to treat
polluted water, it still causes problem in terms of residue. One disadvantage is that
chlorination of residual organic material can generate chlorinated organic compounds
that may also be carcinogenic and harmful to the environment. Residual chlorine or
chloramines may also be capable of chlorinating organic material in the natural
7
aquatic environment. Furthermore, because residual chlorine is toxic to aquatic
species, the treated effluent must also be chemically dechlorinated, adding to the
complexity and cost of treatment.
Chlorination requires high cost compared to Terminalia catappa leaf which is
cheaper since it is easily obtain and the process of the producing tannin extract is
simpler.
Moreover, there are quite a number of Terminalia catappa trees in
Malaysia. Last but not least, the cost to treat residue will be cut off since it produces
lesser residues and is environmentally friendly.
CHAPTER 2
LITERATURE REVIEW
2.1
Introduction
Domestic or sanitary wastewater refers to the liquid discharge from
residences, business buildings and institutions. A human being produces about 0.055
kg of biological oxygen demand (BOD) which is measured at 20°C for five days and
0.08 kg of Suspended Solids (SS) each day and the strength of domestic sewage
depends upon the water used and discharged (Kumar et al., 2006).
Domestic
wastewater can also effect and pollute the environment when the concentration of
nutrients in wastewater is very high.
According to the American College Dictionary (2007), pollution is defined as
to make foul or unclean and dirty. Water pollution occurs when a body of water is
adversely affected due to the addition of large amounts of materials to the water.
When it is unfit for its intended use, water is considered polluted. Two types of
water pollutants exist; point source and non-point source. Point sources pollution
occurs when harmful substances are emitted directly into a body of water. A nonpoint source delivers pollutants indirectly through environmental changes.
An
example of this type of water pollution is when fertilizer from a field is carried into a
9
stream by rain, in the form of run-off which in turn affects aquatic life.
The
technology exists for point sources pollution to be monitored and regulated. Nonpoint sources are much more difficult to control. Pollution arising from non-point
sources accounts for a majority of the contaminants in streams and lakes (Krantz and
Kifferstein, 2005).
According to the Department of Environment (DOE) 2006, the main factor
that contributes to water pollutions are from oxidation pond or Sewerage Treatment
Plant (STP) which consists of a lot of nutrients. Much of the water pollution was
caused by point sources of discharge in the form of sewage and industrial waste
effluents (Kumar et al., 2006). However, the best treatment should be provided to
the treatment facilities and the best method can be applied which is known as “Reuse
Water” in terms of controling the problem.
In fact, the report from Department of Environment (DOE) in 2008 has
shown consistent major pollutants in Malaysian rivers in 2008 where biochemical
oxygen demands (BOD), ammoniacal nitrogen (NH3-N) and suspended solids (SS)
shows high concentration. High BOD was contributed largely by untreated or
partially treated sewage and discharges from agro-based and manufacturing
industries. The main sources of NH3-N were domestic sewage and livestock farming,
whilst the sources for SS were mostly earthworks and land clearing activities. It was
also reported that 74% of polluting agents that mostly polluted our rivers were from
domestic sewage.
2.2
Domestic Wastewater
Wastewater, also termed as sewage effluent can be viewed as used water
originating from domestic premises (toilets, bathrooms, and kitchen-sullage),
10
domestic components of commercial sewage and domestic components of industrial
sewage. Some of this sewage generated from source point is being conveyed via
sewerage systems to treatment facility while some are just discharge directly into
water course such as streams, rivers and lakes. The direct discharge to water courses
however has its significant impact on the environment, in that; it pollutes the water
body leading to outbreak of diseases and foul odour production (Indah Water
Consortium, 2008).
Wastewater has historically been considered as a nuisance to be discarded in
the cheapest and least offensive manner possible. Wastewater temperatures normally
range between 27.5°C and 30°C (Katayon et al., 2006). In general, the temperature
of the wastewater will be higher than that of the water supply. This is because of
addition of warm water from households and heating within the plumbing system of
structure (Mackenzie et al., 2008). Because the number of chemical compounds
found in wastewater is almost limitless, we normally restrict our consideration to a
few general classes of compounds.
Domestic sewage treatment is mainly designed to produce an effluent low in
solids and organic. However, other treatments, which remove the nutrients will alter
the pH or disinfect the effluent may be added depending on the receiving
environment for the effluent. Standards have been established for the quality of
effluent discharged from treatment plants to receiving waters (Table 2.1). These take
the form of acceptable upper limits for various effluent contaminants. Effluents from
treatment plants are regularly sampled and tested in laboratories to ensure that these
standards are being met and that treatment plants are being operated correctly (Indah
Water Consortium, 2008).
11
Table 2.1: Parameter Limits of Effluent of Standard A and B (Environment Quality
Act, 1974)
Parameter
Unit
Standards
A
B
°C
40
40
-
6.0-9.0
5.5-9.0
BOD5 at 20C
mg/l
20
50
COD
mg/l
50
100
Suspended Solids
mg/l
50
100
Mercury
mg/l
0.005
0.05
Cadmium
mg/l
0.01
0.02
Chromium, Hexavalent
mg/l
0.05
0.05
Arsenic
mg/l
0.05
0.10
Cyanide
mg/l
0.05
0.10
Lead
mg/l
0.10
0.5
Chromium, Trivalent
mg/l
0.20
1.0
Copper
mg/l
0.20
1.0
Manganese
mg/l
0.20
1.0
Nickel
mg/l
0.20
1.0
Tin
mg/l
0.20
1.0
Zinc
mg/l
1.0
1.0
Boron
mg/l
1.0
4.0
Iron (Fe)
mg/l
1.0
5.0
Phenol
mg/l
0.001
1.0
Free Chlorine
mg/l
1.0
2.0
Sulphide
mg/l
0.50
0.5
Oil and Grease
mg/l
Not
10.0
Temperature
pH Value
Detectable
In Malaysia, effluent discharges are considered to be one of the main reasons
for water pollution of rivers. Statistics published by the Department of Environment
(DOE) for the year 2007 revealed that 7 river basins are polluted, 45 are still slightly
polluted and the remaining 91 are clean. This showed that there are more clean
12
waters in Malaysia. Therefore, we have to maintain the condition and ensure there
are less polluted rivers. Studies indicate that residential, agricultural and industrial
wastes are three main sources of river pollutions in Malaysia. Degradation of water
quality due to pollution causes adverse effects to aquatic life forms, disturbs the
balance of life and reduces the bioavailability of potable water.
Agriculture still plays a very important role in the development of Malaysia
and a lot of emphasis has been laid on it. The wildlife that thrives abundantly among
us has now been placed in danger. In Cameron Highlands, in the state of Pahang, one
of the many places where vegetables as well as tea which it is famous for, is planted,
human activity has taken it’s toll on the fragile environment (DOE, 2008). Large
scale farming has caused thousands of acres of forest land to be ploughed up and the
habitat of thousands, maybe even millions of wildlife has been destroyed. Some
wildlife flees or migrates to escape the dangers and activities of man. Unknown to
them, they cause an imbalance in their ecosystem, making some areas too densely
populated with predators and not enough food to go around.
Pesticides used in agriculture also play a main role in the degradation of the
environment. Many of these pesticides contain non biological ingredients and can
cause abnormal changes in any wildlife that comes across it (DOE, 2008). In other
words, these chemicals can cause wildlife to mutate. Not only insects to which the
pesticides are aimed towards are affected, but also the animals which feed on them
and they can eventually end up in human bodies. Pesticides pollute the earth; making
it useless as well as poisonous after all the nutrients have been sapped out from it
(DOE, 2008). Thus the land may lay barren and empty for years before it is able to
recover its normal pH level and nutrients. Pesticides also flow into rivers and streams
and eventually seas, causing pollution as it continues its seaward journey.
As Malaysia is fast becoming an industrial country, many rivers in Malaysia
have become polluted due to the many wastes that flow out into the rivers. The
rivers are being used as an outlet for the chemicals to drain away, in turn harming the
13
waters and the lives that revolve around them. Rivers have also become tourist
attraction and this has prompted the construction of hotels and resorts around the
area.
As a result, many of the forests surrounding the river areas have been
chopped down. The surrounding soil has no roots to hold onto and soon erode when
the rains come. The soil runs into the rivers and soon the rivers become murky and
block all the sunlight from reaching the aquatic life in the rivers and streams (DOE,
2008). A good example is the construction of a new golf course near the waterfall at
tourist attraction Fraser’s Hill in the state of Pahang, causing it to become extremely
murky and dirty due to silt and sand that comes from the construction (DOE, 2008).
The waterfall which was the centrepoint of the hill has now lost all its attraction just
because of the overwhelming need to attract more tourists to the place by building
more facilities.
The burden on rivers to supply fresh water is likely to increase as demand is
growing at 4% annually and is projected to reach 20 billion m3 by the year 2020.
Since rivers form 97% of our fresh water resource, this is an indication that water
supply would have to be treated extensively in future and the cost would have to be
absorbed by the public. This fact alone is enough to give us a wake up call on the
need for careful water quality monitoring to keep our rivers clean. Under these
circumstances, river pollution problem can be greatly improved if domestic effluents
discharges can be reduced by applying it to other beneficial reuses (Mohd Ismid et
al., 2003).
It is expected that the increase in population growth can lead to
introduction of more human activities and this results in more pollution being caused.
Viewed in the aspect of tourists into the country, more activities are created and
pollution of water bodies becomes more pronounced. Thus, sources of wastewater
depend on the activities in the area.
14
2.3
Charactersitics of Domestic Wastewater
Domestic wastewater is defined as used-water and includes substances such
as human waste, food scraps, oils, soaps and chemicals. Domestic wastewater
includes used-water from sinks, baths, toilets, washing machines and kitchen
(Jabatan Pembentungan Sarawak, 2008). Domestic wastewater could severely affect
and deteriorate in rivers and coastal waters. The impacts of untreated or partially
treated of wastewater include the unsightly littering of the rivers, can cause foul
smells and potential health hazard. Continuous pollution may threaten the survival of
aquatic life in rivers.
There are two types of domestic wastewater namely black water (wastewater
from toilets) and grey water (waste from kitchen, bathroom, urine and sink). Black
water and gray water have different characteristics, but both contain pollutants and
disease-causing agents that require treatment. Some areas in the U.S., including
Arizona (Direct Reuse of Reclaimed Water Rule, effective 01/16/01), permit the use
of innovative systems that safely recycle household gray water for reuse in toilets or
for irrigation to conserve water and reduce the flow to treatment systems. Table 2.2
is the list of the composition for typical untreated domestic wastewater with their
typical and range of the concentrations. .
The characteristic of generated wastewater depends on the landuse of the
catchments area.
The landuse can range from institutional, commercial, and
industrial to household, of which wastewater generated from each activity has its
characteristics such as the key sewage pollution measures (biochemical oxygen
demand (BOD), and suspended solids(SS)) and their corresponding indicators
(ammonia (NH3) and E. coli).
15
Table 2.2: Composition of Typical Untreated Domestic Wastewater (Burks and
Minnis. 2006)
Constituent
Unit
Range
Typical
Total Solids
mg/L
300-1200
700
Dissolved Solids
mg/L
250-850
500
Suspended Solids
mg/L
100-400
220
Volatile Solids
mg/L
70-300
150
BOD5
mg/L
100-400
250
TOC
mg/L
100-400
250
COD
mg/L
200-1,000
500
Total Nitrogen
mg/L
15-90
40
Ammonia
mg/L
10-50
25
Total Phosphorous
mg/L
5-20
12
Chloride
mg/L
30-85
50
Sulphate
mg/L
20-60
15
Grease
mg/L
50-150
100
Total Coliform
colonies/100 mL
106-108
107
VOCs
µg/L
100-400
250
2.3.1
Physical Characteristics
Fresh, aerobic, domestic wastewater has been said to have the odor of
kerosene or freshly turned earth (Metcalf and Eddy, 2004). Aged, septic sewage is
considerably more offensive to the olfactory sense. Fresh sewage is typically gray in
colour. Septic sewage is black. One cubic meter of wastewater weighs approximately
1000000 g and will contain about 500 g of solids. One-half of the solids will be
dissolved solids such as calcium, sodium and soluble organic compounds. The most
16
important physical characteristic of wastewater is its total solids content, which is
composed of floating matter, colloidal matter, and matter in solution. Other
important physical characteristics include particle size distribution, turbidity, color,
temperature. Odor sometimes considered a physical factor, is considered in the
following:
i) Total Solids (TS)
Solids usually measured in TS (Total Solids) and the definition of solids
only suitable used for the residue that remain after a wastewater sample have
been evaporated and dried at a specified temperature (103°C-105°C).
Suspended matter consists of silt, clay, fine particles of organic and inorganic
matter, soluble organic compounds, plankton and other microscopic organism
(Metcalf and Eddy, 2004). .
Measurement of suspended matter transport is particularly important where it
is responsible for pollutant transport. Usually sediment concentration and
load increase exponentially with discharge. Particles may settle or
resuspended under different discharge condition. Particles vary in sizes from
approximately 10 nm in diameter to 0.1 mm in diameter, although it is
usually accepted that suspended matter is the fraction that will not pass
through a 0.45 µm pore diameter film (Metcalf and Eddy, 2004).
ii) Turbidity
Turbidity, a measure of light-transmitting properties of water, is another test
used to indicate the quality of waste discharges and natural waters with
respect to colloidal and residual supended matter.
The measurement of
turbidity is based on comparison of the intensity of light scattered by a
sample to the light scattered by a reference suspension under the same
17
conditions (Standard Methods, 2005). The turbidity measurements are
reported as NTU (Nephelometry Turbidity Unit) or FTU (Formazing
Turbidity Unit) (Metcalf and Eddy, 2004).
iii) Temperature
Temperature is one important parameter in studying wastewater and it is
measured in degrees celcius (°C) or farenheit (°F) unit (Metcalf and Eddy,
2004). The temperature of wastewater is commonly higher than that of the
local water supply, because of the addition of warm water from households
and industrial activities. Depending on the location and time of year, the
effluents temperature can be either higher or lower than the corresponding
influent values. The temperature of water is a very important parameter
because of its effect on chemical reactions and reaction rates. It also can
effect the DO concentration where oxygen is less soluble in warm water than
in cold water.
The increase in the rate of biochemical reactions that
accompanies an increasre in temperature, combined with the decrease in the
quantity of oxygen present in surface waters, can often cause the decrease in
the quantity of oxygen present in surface waters (Metcalf and Eddy, 2004).
Temperature is one of the environmental factors that determine which
organisms will thrive and which will diminish in numbers and sizes. Heat
input into aquatic systems and the resultant temperature significantly affects
the biological community and the beneficial use of the waters systems
(Metcalf and Eddy, 2004). Temperature conditions may be either unnatural
highs or lows or they may be because of natural seasonal and daily
temperature fluctuations.
Temperature problems may also develop in
industrialized nations as a result of the use of the water resource for cooling
purposes. Changes in temperature can also affect both fish and lower forms of
aquatic life as each organism has it range of temperature for optimum growth
as well as lethal level. It also affects the usability of water for beneficial
purposes (Marks, 2001).
18
iv)
Taste and Odor
Taste and odor is the one of the parameter which is subjective in
measurement of wastewater and it is totally different depending on the types
of wastewater (Metcalf and Eddy, 2004). Wastewater usually not regardless
from the taste and odor disturbance because from the organic waste such as
fenol and chlorofenol in human waste.
Odor is a volatilized chemical
compound, generally at a very low concentration, which humans and other
animals perceive by the olfactory sense. Odors are also called smells, which
can refer to both pleasant and unpleasant odors. Assessment of odour is
usually not included in the water quality assessment. If a change in odour is
detected, it might indicate water quality problem that requires further
investigation.
v) Color
Fresh wastewater usually is light brownish-gray color. However, as the travel
time in the collection system increases, and more anaerobic conditions
develop the color of wastewater changes sequentially from gray to dark gray,
and ultimate to black (Metcalf and Eddy, 2004).
2.3.2
Chemical Characteristics
Chemical attributes of a waterway can be important indicators of water
quality. Chemical attributes of water can affect aesthetic qualities such as how water
looks, smells, and tastes. Chemical attributes of water can also affect its toxicity and
whether or not it is safe to use. Since the chemical quality of water is important to the
health of humans as well as the plants and animals that live in and surrounding
streams, it is necessary to assess the chemical attributes of water.
Commonly
19
measured chemical parameters include BOD, COD, AN, pH and DO. Table 2.3
shows that the typical composition of untreated domestic wastewater.
i) Biological Oxygen Demand (BOD)
When biodegradable organic matter is released into a watercourse,
microorganisms feed on them and break them down into simpler substances
(Metcalf and Eddy, 2004). With oxygen present when the decomposition, the
process is said to be in an aerobic environment and this process produces
non-objectionable, stable end products such as carbon dioxide (CO2),
sulphate (SO4) and nitrate (NO3). Processes without the presence of oxygen
are said to be in an anaerobic environment.
Therefore, the biological oxygen demand defined as the amount of oxygen
required by microorganisms to oxidise organic wastes aerobically i.e. in the
presence of oxygen. It is often expressed in milligrams of oxygen required
per litre of wastewater (mg/L) although it may have various units. Most
pristine rivers will have a 5-day BOD below 1 mg/l. Moderately polluted
rivers may have a BOD value in the range of 2 to 8 mg/l.
The total amount of oxygen that will be required for biodegradation is an
important measure of the impact that a given effluent will have on the
receiving watercourse.
A standard practice to measure and report the
depletion of oxygen demand had been restricted to a five-day period and the
test is run at a fixed temperature of 20°C. The five-day BOD, also known as
BOD5, is the total amount of oxygen consumed by microorganisms during the
first five days of the biodegradation process.
20
ii) Chemical Oxygen Demand (COD)
The chemical oxygen demand (COD) test is commonly used to indirectly
measure the amount of organic compounds in water. Most applications of
COD determine the amount of organic pollutants found in surface water (e.g.
lakes and rivers), making COD a useful measure of water quality. It is
expressed in milligrams per liter (mg/L), which indicates the mass of oxygen
consumed per liter of solution. Older references may express the units as
parts per million (ppm).
The COD test is more sophisticated and has more advantages than the BOD5
test because the results are available within two and half hours instead of five
days. The test also has the capability of measuring organic material which is
resistant to biological decay.
The COD is used comprehensively in
evaluating the waste treatment processes for industrial wastes for which
BOD5 test is not applicable or for which more rapid data is needed. For the
test, the usage of strong oxidizers under standards such as permanganate,
dichromate, or periodate oxidizes the organic compound (Metcalf and Eddy,
2004).
21
Table 2.3: Typical Composition of Untreated Domestic Wastewater (Metcalf and
Eddy, 2004)
Constituents
Total Solids (TS)
Total Dissolved Solids, TDS
Fixed
Total Suspended Solids, TSS
Fixed
Seattleable Solids
Concentration
Unit
mg/l
mg/l
mg/l
mg/l
Biological Oxygen Demand
BOD5, 20°C
Weak
Medium
Strong
350
720
1200
250
500
850
145
300
525
100
220
350
20
55
75
5
10
20
110
220
400
Total Organic Carbon, TOC
mg/l
80
160
290
Chemical Oxygen Demand, COD
mg/l
250
500
1000
Nitrogen (Total N)
20
40
85
Organic
8
15
35
12
25
50
Nitrite
0
0
0
Nitrate
0
0
0
Phosphorus (Total P)
4
8
15
1
3
5
3
5
10
Ammonia
Organic
mg/l
mg/l
Inorganic
Chloride
mg/l
30
50
100
Sulphate
mg/l
20
30
50
Alkalinity (as CaCO3)
mg/l
50
100
200
Oil and Grease
mg/l
50
100
150
Total Coliform
no/100ml
106-107
107-108
107-108
22
iii) Dissolved Oxygen (DO)
The amount of dissolved oxygen present in a watercourse is one of the most
important criteria to measure water quality (Metcalf and Eddy, 2004). It is
also commonly used as an indicator of a river’s health. The number of life
forms that survive begins to decrease as the level of DO drops below 4 mg/L.
In extreme cases, when anaerobic condition exists, most high forms of life are
either killed or driven off. Eventually conditions like floating sludge,
bubbling, odorous gasses and slimy fungal growths will manage to survive.
Concentrations in unpolluted waters are usually close to or less than 10 mg/L.
Concentrations below 5 mg/L may adversely affect the functioning and
survival of biological communities and below 2 mg/L may lead to the death
of most fish (Metcalf and Eddy, 2004). There are various factors that affect
the amount of DO available in a river. Oxygen demanding wastes reduces DO
level while photosynthesis process further adds DO during the day but
removes oxygen in the night. The respiration of the life forms in the water
body also reduces the DO level. Besides that, tributaries also bring in their
own oxygen supply that mixes with the main river.
iv) pH
pH is an important variable in water quality assessment as it influences many
biological and chemical processes within a water body and all processes
associated with water supply and treatment. pH is a measure of the hydrogen
ion concentration or activity, [H+] (Metcalf and Eddy, 2004). The pH value
is the negative normal logarithm of the hydrogen ion activity (mol/L) and has
a value of 7 at 25oC in pure water (neutral point). The pH value can increase
or decrease because of the presence of acids (decrease) or alkali (increase)
and the hydrolysis of dissolved salts.
23
2.3.3
Biological Characteristics
Biological attributes of a waterway can be important indicators of water
quality. Biological attributes refer to the number and types of organisms that inhabit
waterways. The poorer water quality of water, the fewer the number and types of
organisms that can live in it. Table 2.4 and Table 2.5 show the typical of organisms
and microorganisms that contains in untreated domestic wastewater.
i) Pathogens
The most common human microbial pathogens found in recycled water are of
enteric in origin. Enteric pathogens enter the environment in the faeces of
infected hosts and can enter water either directly through defecation into
water, contamination with sewage effluent or from run-off from soil and
other land surfaces (Feachem et al., 1983).
The types of enteric pathogens that can be found in water include viruses,
bacterias, protozoas and helminths. The risk of water-borne infection from
any of these pathogens can be reliant on a range of factors including
pathogen numbers and dispersion in water, the infective dose required and
the susceptibility of an exposed population, the chance of fecal contamination
of the water and amount of treatment undertaken before potential exposure to
the water (Haas et al., 1999).
ii) Viruses
Enteric viruses are the smallest of the pathogens found in water. They are all
obligate intercellular parasites that require the infection of host cells of a
suitable host and then force the host cell to produce multiple copies of the
virus (Toze, 1997). This lack of ability to self-replicate means that viruses
24
are present in water as inactive particles. The majority of these viruses can
be commonly detected in fecal contaminated water, for example sewage
effluent; however, the wild type of poliovirus has been eradicated in
developed nations such as Australia due to widespread vaccination (Ward et
al., 1993). Most enteric viruses have a narrow host range meaning that most
viruses of interest in recycled water only infect humans (Haas et al., 1999).
This means that only human fecal contamination of water need to be
considered a concern for viral infection of humans. Conversely, water borne
human viruses are rarely a problem for other animals.
iii)
Bacteria
Bacteria are the most common microbial pathogens found in recycled waters
(Toze, 1999).
There are a wide range of bacterial pathogens and
opportunistic pathogens which can be detected in wastewaters. Many of the
bacterial pathogens are enteric in origin; however, bacterial pathogens which
cause non-enteric illnesses (e.g. Legionella spp., Mycobacterium spp., and
Leptospira) have also been detected in wastewaters (Wilson and Fujioka
1995). Bacterial pathogens are metabolically active microorganisms that are
capable of self-replication and are therefore, theoretically capable of
replicating in the environment.
In reality, however these introduced
pathogens are prevented from doing so by environmental pressures (Toze and
Hanna, 2002). Like other enteric pathogens, a common mode of transmission
is via contaminated water, food and by direct person to person contact (Haas
et al., 1999).
iv) Protozoa
Enteric protozoan pathogens are unicellular eucaryotes which are obligate
parasites. Outside of an infected host they persist as dormant stages known
as cysts or oocysts. There are several protozoan pathogens which have been
25
isolated from wastewater and recycled water sources (Gennaccaro et al.,
2003).
The most common detected are Entamoeba histolytica, Giardia
intestinalis (formerly known as Giardia lamblia), and Cryptosporidium
parvum (Toze, 1997).
Infection from all three of these protozoan pathogens can occur after
consumption of food or water contaminated with the (oo)cysts or through
person to person contact (Carey et al., 2004). Giardia and Cryptosporidium
are ubiquitous in fresh and estuarine waters and have been detected in
numerous countries around the globe (Ferguson et al., 1996). E. histolytica
can be detected in all parts of the world, although it is more prevalent in
tropical regions (Feachem et al., 1983).
Similar to bacterial pathogens,
various domestic and wild animals can be a source of these protozoa and be
infected by them.
2.3.4
Nutrients
A major contaminant type commonly found in wastewater is organic and
inorganic nutrients. The most common organic nutrient is dissolved organic carbon
(DOC). DOC can take various forms depending on the source of the wastewater
(Kumar et al., 2006). The source of the organic carbon can also influence the
bioavailability of the nutrient. For example DOC from drainage water would be
likely to be more recalcitrant than DOC present in the effluent from a sewage
treatment plant or from a food processing plant. It has been noted that the organic
carbon present in recycled water can stimulate the activity of soil microorganisms
(Ramirez-Fuentes et al., 2002).
26
Table 2.4: Typical of Organisms of Untreated Domestic Wastewater (Metcalf and
Eddy, 2007)
Types of Pathogen
Disease
Counter in
Wastewater
Infection
Dose
Virus
Enteroviruses
Poliovirus
Poliomylitis
Enterovirus
Gastroenteritis, heart
Echovirus
anomalies, meningitis
Coxsackievirus
Hepatitis A virus
Adenovirus
Hepatitis
Respiratory disease,
conjunctivitis
Reovirus
Not clearly established
Callcivirus
Gastroenteritis,
Norwalk agent
Diarrhoea, vomiting,
SSRV
fever
Rotavirus
Gastroenteritis
Astrovirus
Gastroenteritis
Medium to
High
Low
Bacteria
Vibrio cholerae
Salmonella typhi
Enteropathogenis E. coli
Campylobacter jejunei
Shigella dysinterae
Yersinia enterocolitica
Cholera
High
Typhoid, Salmonellosis
Gastroenteritis
Medium to
Gastroenteritis
High
Dysentery
High
High
High
Low
Yersiniosis
High
Protozoa
Giardia intestinalis
Giardiasis
Cryptosporidium parvum
Diarrhea, fever
Entamoeba histolytica
Amoebic dysentery
Medium to
Low
High
Low
Low
27
Table 2.5: Typical and Number of Microorganisms of Untreated Domestic
Wastewater (Metcalf and Eddy, 2007)
Organisms
Counter (No./mL)
Total Coliform
105-106
Fecal coliform
104-105
Fecal streptococci
103-104
Enterococci
102-103
Shigella
100-103
Pseudomonas aeroginosa
100-102
Clostridium perfringens
101-102
Mycobacterium tuberculosis
101-103
Protozoan cysts
100-101
Giardia cysts
101-103
Cryptosporidium cysts
10-1-102
Helminth ova
10-2-101
Enteric virus
101-102
Magesan et al., (2000) noted that the organic and inorganic nutrients in
treated effluent that had a high carbon to nitrogen ratio stimulated the soil
microorganisms which, in turn, decreased the hydraulic conductivity of the irrigated
soil. The microorganisms in this study reduced the hydraulic conductivity in the soil
by excess cell growth and the production of biofilm structures, both of which would
have clogged up the pore spaces between the soil particles. In a different study,
28
Sheikh et al., (1987) observed that salad crops irrigated with the treated effluent,
which had raised concentrations of inorganic nutrients, produced higher yields than
similar crops irrigated with groundwater. Chakrabarti (1995) observed that rice crops
gave a higher yield when irrigated with raw or partially diluted sewage effluent
compared to unlamented groundwater.
In the same study Chakrabarti (1995) noted that initially the rice did better if
a fertilizer was used in conjunction with the sewage effluent but this requirement for
additional nutrients decreased over time due to accumulation of nutrients,
particularly nitrogen, in the soil. Evidently, while the additional nutrients can be a
bonus as additional fertilizer, excess nutrients, particularly carbon and nitrogen, can
have an adverse effect through excessive microbial activity and growth. Thus, care
needs to be taken in the concentrations of nutrients in the recycled water to avoid
detrimental impacts on soil porosity.
2.4
Oxidation Pond (OP)
Oxidation Ponds (or Stabilization Ponds) are a popular sewage treatment
method for small communities because of their low construction and operating costs
(Indah Water Consortium, 2008). Oxidation ponds represent 12% (500 numbers) of
all sewage treatment plants. New oxidation ponds can treat sewage to Standard B
effluent level but require maintenance and periodic desludging in order to maintain
the standard. OPs may comprise one or more shallow ponds in a series. The natural
processes of algal and bacteria growth exist in a mutually dependent relationship.
According to Indah Water Consortium, 2008, oxygen is supplied from natural
surface aeration and by algal photosynthesis. Bacteria present in the wastewater use
the oxygen to feed on organic material, breaking it down into nutrients and carbon
29
dioxide. These are in turn used by the algae. Other microbes in the pond such as
protozoa remove additional organic and nutrients to polish the effluent. There are
normally at least two ponds constructed.
The first pond reduces the organic material using aerobic digestion while the
second pond polishes the effluent and reduces the pathogens present in sewage.
Sewage enters a large pond after passing through a settling and screening chamber.
After retention for several days, the flow is often passed into a second pond for
further treatment before it is being discharged into a drain. Bacteria already present
in the sewage acts to break down organic matter using oxygen from the surface of the
pond. Figure 2.6 shows the standards effluent of domestic wastewater from oxidation
pond before discharge to the rivers. OPs require large amounts of land and the
degree of treatment is weather dependent. They are incapable of achieving a good
standard of effluent consistently. It is this variation in performance, which requires
the gradual phasing out of this type treatment plant.
Table 2.6: Effluents of Oxidation Pond (Indah Water Consortium, 2008)
Compounds
Biological Oxygen Demand
Raw Sewage
(mg/L)
Effluent
(mg/L)
DOE Standard B
(mg/L)
200-400
20-100
50
200-350
30-150
100
(BOD)
Suspended Solids (SS)
Existing Sewage Treatment Plant (STP) in Malaysia which are mostly
oxidation ponds have always faced with problem attaining the required Department
of Environment (DOE) effluent standards.
Major problems related to odor
emissions, instability in treatment process and excess sewage production persisted
until today.
30
2.5
Wastewater Reuse Applications
In the planning and implementation of water reclamation and reuse, the
reclaimed water application will usually govern the wastewater treatment needed to
protect public health and the environment, and the degree of reliability required for
the treatment processes and operations (Kumar et al., 2006). Because of health and
safety concerns, water reuse applications are mostly restricted to non portable uses
such as landscape and agricultural irrigation. Table 2.7 shows the advantages and
disadvantages of wastewater reuse applied in Malaysia.
The reuse of water is just one source of water that has potential for use in an
agricultural setting. Reused water does, however, have a major advantage in that it is
usually a constant and reliable supply, particularly with sources such as treated
sewage effluent or industrial discharges. As well as being a constant source of water,
many waters suitable for reuse are produced in large volumes, which if not used
would be merely discharged into the environment. It is well known that discharge of
effluents, treated or non-treated, into the environment, particularly natural water
bodies such as lakes, rivers and the coastal marine environments can cause severe
degradation of these water ways (Kumar et al., 2006).
The degradation is often related to the presence of organic and inorganic
nutrients which can cause problems such as eutrophication and algal blooms.
Reusing these discharged effluents can have a significant impact on reducing or
completely removing the impact of these effluents from receiving environments. In
addition, the reuse of wastewaters for purposes such as agricultural irrigation reduces
the amount of water that needs to be extracted from environmental water sources
(Gregory, 2000; USEPA, 1992).
31
Wastewaters can often contain significant concentrations of organic and
inorganic nutrients for example nitrogen and phosphorus. There is potential for these
nutrients present in recycled water to be used as a fertilizer source when the water is
recycled as an irrigation source for agriculture. Soil microorganisms have been
observed to have increased metabolic activity when sewage effluent is used for
irrigation (Ramirez-Fuetes et al., 2002; Meli et al., 2002).
Table 2.7: Advantages and Disadvantages of Wastewater Reuse (Kumar et al., 2006)
Advantages
Disadvantages
•
Conserves water
•
Low-cost method for sanitary
communities with prolonged
disposal of municipal wastewater
contact to untreated wastewater
Reduces pollution of rivers,
and consumers of vegetables
canals and other surface water
irrigated with wastewater
•
resources
•
•
•
Conserves nutrients, reducing the
need for artificial fertilizer
•
Increases crop yields
•
Provides reliable water supply for
Health risks for irrigators and
Contamination of groundwater
(nitrates)
•
Buildup of chemical pollutants in
the soil (heavy metals)
•
Creation of habitats for disease
vectors
farmers
•
Excessive growth of algae and
vegetation in canals carrying
wastewater (eutrophication)
The Kingdom of Saudi Arabia, being an arid country, lacks perennial rivers.
Groundwater constitutes the main source of the natural water supply of which
agricultural irrigation consumes a major share. The rainfall in the Kingdom is low
and therefore recharge of the deep sedimentary aquifers is mostly insignificant.
32
Because of extraction of groundwater to meet the demand in various sectors; the nonrenewable aquifers are showing signs of significant water level decline, and
wastewater effluent can supplement these demands to a certain degree in non
domestic sectors.
The Kingdom's policy is to utilize all available treated municipal wastewater
in a most beneficial manner for several purposes, among which the agricultural
sector is afforded top priority. The importance of reclaimed wastewater is due to the
great need for new water resources to meet the increasing water demands for
agriculture and landscape irrigation, for industrial abstraction and for possible
recharging of aquifers (Kalthern et al., 1985).
Suitable water resources for agricultural development in Kuwait are very
limited indeed. Consequently, the availability of irrigation water at a reasonable cost
is one of the most important factors to be considered for the development of
agriculture. Desalinated seawater used in houses is rather expensive for direct use
for irrigation. Hence, it was decided to reuse wastewater (Cobham, 1985). Untreated
sewage effluent has been used to irrigate forest trees far away from inhabited areas
for a long time. Until the late 1970s, agriculture in Kuwait was severely limited. The
total cropped area in 1976 amounted to only 732 ha.
The country relied heavily on food imports and the imports of both fresh and
dried are considered unnecessarily high. In 1985 a new area of 700 ha was added to
this farm using the same source of irrigation water and irrigation method, making the
total area 1600 ha (Cobham, 1985). About 27 million m3/year of treated effluent are
currently being used, which will rise to 125 million m3/year by the year 2010,
constituting one of the most ambitious schemes for agricultural water reuse in the
Middle East.
33
2.6
Potential Risks from Using Recycled Water
There have been a number of risk factors identified in using reused waters for
purposes such as agricultural irrigation. Some risk factors are short term and vary in
severity depending on the potential for human, animal or environmental contact (e.g.
microbial pathogens), while others have longer term impacts which increase with
continued use of recycled water (e.g. saline effects on soil).
2.7
Agricultural Reuse
The introduction of wastewater system for irrigation indicates a progressive
step but it is generally possible to limit or dispense with utilization of the fertilizing
constituents of sewerage or sewage effluents in order to protect the public health.
Accordingly, most health authorities prohibit the irrigation of vegetables, garden,
berries or fruits with partially treated or undisinfected sewage (Kumar et al., 2006).
The quality of irrigation water is of interest in relation to wastewater disposal by
irrigating agricultural areas either by direct discharge from the drainage system or by
diversion of sewage polluted receiving waters.
Among the types of wastewater reuses, landscape irrigation was the earliest
application and receives most attention. It can be used in parks, school yards, golf
courses, cemeteries, home lawns (Kiely, 1997). Water quality considerations for
landscape are also very similar to those for agricultural irrigation in order to protect
public health and to avoid becoming a public nuisance. The degree of treatment
needed for the applications also varies depending on irrigation methods, which can
be divided to drip or sprinkler irrigation, depending on the potential for public access
(Jones, 1993). Landscape watering using effluent is also known to be beneficial and
advantageous since it provides low cost water source, eliminates the use of synthetic
34
fertilizer (Angelakis et al., 1998), and eradicate the use of expensive yet complicated
tertiary treatment (Alhomoud et al., 2002).
According to Indian Standards, in order to regulate disposal of effluents on
land, it is necessary to limit certain constituents in effluents, especially those
considered toxic, so that the effluent may comply with normally accepted irrigation
water quality. The standards applicable for land application of wastewater in India
as per Environment (Protection) Act, 1986 are shown in Table 2.8.
Irrigation is an excellent use for sewage effluent because it is mostly water
with nutrients. The use of sewage effluents for irrigation of agricultural crops is an
attractive and popular wastewater reuse option for the following reasons:a)
Where crops need to be irrigated, water tends to be scarce, and
wastewater can supplement available freshwater resources.
(b)
Irrigated agriculture requires large amounts of water which are used
only once, since irrigation basically is a consumptive use;
consequently, irrigation water requirements represent a major portion
of the total water demand in dry areas.
(c)
Agriculture can beneficially use not only the water, but also, within
certain limitations, the additional resources found in wastewater, such
as organic matter, nitrogen, phosphorus, potassium, minor elements,
and other nutrients.
(d)
Irrigation is relatively flexible with respect to water-quality
requirements: Some crops may be irrigated with low-quality water
without major risks, and some water quality problems can be
overcome by suitable agronomic practices.
35
Table 2.8: Standards of Treated Wastewater for Irrigation in India (Kumar et al.,
2006)
No
Parameter
Limits*
1
Colour and Odour
**
2
Suspended Solids
200
3
Total Disoolved Solids
2100
4
pH Values
5.5 to 9.0
5
Oil and Grease
10.0
6
BOD5 at 20°C
100
7
Arsenic (As)
0.2
8
Boron (B)
2.0
9
Percent Sodium
60
10
Residual Sodium Carbonate
5.0
11
Cyanide (Cn)
0.2
12
Chloride (Cl)
600.0
13
Sulphate (SO4)
1000
14
Pesticides
Absent
* All values are maximum limit expressed as mg/L except pH
** All efforts should be made to remove colour and unpleasant odour as far
as possible
A distinction should be made between two types of irrigation using
municipal effluent: restricted and unrestricted irrigation (Ayers et al., 1981). The
concept of restricted irrigation refers to the use of a low-quality effluent only in
specific agricultural areas and for specific crops. Restrictions imposed in connection
with this reuse category are related not only to the type of crops to be cultivated, but
36
also to the type of soils to be irrigated, the location of irrigated fields with respect to
potable aquifers, irrigation methods, crop-harvesting techniques, fertilizer application
rates, distance of irrigated fields to roads and houses, distance between pipelines
carrying potable and non-potable water. The "restricted irrigation" concept has the
advantages of simplicity and low treatment cost, but it is generally applicable only to
small amounts of wastewater to be used in specific locations, where areas to be
irrigated and crops to be cultivated are well-defined and unlikely to change.
Restricted irrigation of large areas is technically feasible, but problems
increase dramatically as the size of the system increases. However, a few large
systems (notably at Melbourne, Australia) have also been constructed. Farmers are,
in general, unwilling to accept a low-quality effluent in equal exchange for fresh
water. Also, farmers and farm personnel will have to be trained to properly handle
the effluent. Irrigation techniques should be selected that minimize contact between
irrigators and wastewater. Aerosols can result in transport of microorganisms to a
considerable distance from the field, especially if the effluent is applied with
sprinklers (Idelovitch and Bouwer, 1984). Accidental ingestion of the low-quality
water and the accidental use of the water to irrigate vegetables or other crops brought
raw into the kitchen represent additional health hazards.
2.8
Sustainable Environment and Development
Once a limited regulatory constraint, environment issues have become a
profound mainstream concern that impacts on every business, government,
organisation and individual on the planet. These issues are so important that they
will increasingly dominate the economic, political and social agenda of the 21st
century and will be the driving force of new industrial revolution (Energy & Climate
Change 2007). In its 2007 report the UN’s Intergovernmental Panel on Climate
Changes (IPCC) found that unless global warming is dealt within the next 10-15
37
years it will lead to catastrophic consequences. Climate change is global in nature
and only coordinated international actions can resolve it. Climate change, resource
availability, waste disposal and pollution are linked to sustainability.
Sustainable development is development that meets the needs of the present
without compromising the ability of future generations to meet their needs. It is the
balance of three interconnected dimensions defined by The Declaration of Rio on
Environment and Development in 1992 as environmental protection, economic
growth, and social development (Energy & Climate Change 2007). Sustainable
development calls for long-term changes in patterns of production and consumption.
These changes are being driven by international intergovernmental agreements that
will impact on all businesses and individuals. The Kyoto treaty requires emissions of
GHG to be reduced to their 1990 levels by 2012. The 2020 European target is a 20%
reduction compared to 1990 and for 2050 a 75% reduction.
There appears to be a growing consensus on environmental economic
implications. A more recent IPCC report identifies that the most stringent mitigation
target would reduce global growth by 0.12% per year to 2050; it could be less.
Improved energy efficiency is the fastest and cheapest way to reduce CO2 because
investment in available technologies would cut carbon emissions by about half of the
amount needed to stabilize them. There are two other realities concerning energy i.e.
supply is limited and expensive and the cheapest kW of energy is the one not used.
The Declaration of Rio on Environment and Development in 1992 (Energy &
Climate Change 2007) recognised that sustainable development was a balance of
three dimensions i.e. environmental protection, economic growth and social
development. Economic, social and environmental processes are interconnected.
Neither public nor private entities can work in isolation on a single dimension
because their actions must take into consideration the interplay of them all.
Sustainable development goes beyond environmental preservation. Economic
prosperity and solidarity is required by society to satisfy its material and non-
38
material needs. Sustainability should also contribute to the bottom line performance
of business. Sustainable development calls for long term changes in patterns of
production and consumption. The aim is to protect the environment and its resources
while at the same time satisfying human needs and boosting progress.
Global
interdependencies also need to be balanced. The implications of today’s actions
must be considered to ensure that future generations are able to satisfy their needs.
2.9
Global Warming and Climate Change
The primary cause of climate change is greenhouse gases (GHG) produced by
a wide range of human activities including agriculture, transport, sewage treatment
and energy generation. There is a direct correlation between CO2 (carbon) fossil
emissions and energy consumption. Carbon footprint is being used in CO2 abatement
efforts as an indicator to quantify the global warming potential (GWP) of total
greenhouse gases incurred throughout the life cycle and associated supply chains.
The global population is facing what is very possibly the biggest challenge it
has ever confronted. As professional technologists, scientists, engineers and
environmentalists, however we are effectively the front line of the human response to
these challenges, and we have a responsibility to use our technology, skills and
resources to ensure that today’s projects are constructed using methods that will ease
rather than exacerbate the situation, and that tomorrow’s projects are designed to
conform with best environmental practice. Climate change will influence buildings
and other construction projects in two ways (IPCC, 2007).
The nature of climate change is global. It makes no difference if greenhouse
gases (GHG) are emitted in Buenos-Aires, Melbourne, Paris or Chicago. In the same
way, no part of the planet is immune to the effects of climate change (IPCC, 2007).
39
The greenhouse effect is where most infrared radiation is absorbed by the
atmosphere and reemitted in all directions by greenhouse gas molecules and clouds.
The effect of this is to warm the earth’s surface and lower atmosphere (Figure 2.1).
The Kyoto treaty formally identifies six greenhouse gases (CO2, CH4, N2O, HFC,
PFC and SF6). These are produced by a wide range of human activities, including
agriculture, transport, sewage treatment and energy generation. There is a direct
correlation between CO2 (carbon) fossil emissions and energy consumption. The
greenhouse effect is where most infrared radiation is absorbed by the atmosphere and
re-emitted in all directions by greenhouse gas (GHG) molecules and clouds as shows
in Figure 2.2.
Figure 2.1: The increasing concentration of greenhouse gases from the year 0 to
2005 (Sources; IPCC, 2007)
Firstly, regulatory measures instigated by governments to reduce greenhouse
gas emissions will mean that we have to make fundamental changes to the way that
energy is generated, and to the way that it is delivered and consumed by the building
and industrial sectors. Actions to reduce greenhouse gas emissions are referred to as
‘mitigation measures’. These measures not only reduce greenhouse gas emissions,
40
but they also provide an opportunity to take advantage of a greater diversity of
energy sources, including those with a smaller carbon footprint. Secondly, the
climate will continue to change for at least the next 50 years. Consequently, new
buildings will have to be constructed to different performance parameters, and
existing buildings and infrastructure will have to be adapted to function effectively
under a different range of climatic conditions (IPCC, 2007).
Figure 2.2: The greenhouse effect is where most infrared radiation is absorbed by
the atmosphere and re-emitted in all directions by greenhouse gas (GHG)
molecules and clouds (Source: IPCC, 2007)
In coastal areas, for example, buildings may have to be adapted to cope with
rising sea levels. Adjusting to climate change is termed ‘adaptation’. To put the
challenge of carbon reduction into perspective, it has been predicted that, if present
policies remain unchanged, world energy demand will increase by over 50% between
now and 2030, and energy-related CO2 emissions will climb by 52%. These trends
imply that there must be a very significant drive to improve energy efficiency, and to
move to sources of energy that result in lower atmospheric emissions of greenhouse
gases (IPCC, 2007).
41
Global warming pollution can also be reduced by improved agricultural soil
and manure management, which produces important water quality benefits as well
(IPCC, 2007). According to recent estimates, agriculture accounts for about 6 percent
of all global warming pollution in the United States. Nitrous oxide is the most
significant greenhouse gas emitted through agricultural production. Agricultural soil
management activities account for 78 percent of nitrous oxide emissions, most of
which result from the application of nitrogen fertilizers to cropland, which in turn
causes water pollution. Currently, U.S. farmers apply about 20 to 30 percent more
nitrogen fertilizer than needed. Scientists further estimate that reducing nitrogen
fertilizer use would reduce downstream water pollution by more than 20 to 30
percent. Proven nutrient conservation practices are available that can substantially
reduce loss of nitrogen to the atmosphere and to surface and groundwater.
New technologies, recycling, and demand management have been analysed to
reduce the carbon footprint for the whole water cycle. The carbon footprint of water
supply systems has significant impacts on the carbon footprint of final products.
Water supply system is an essential utility required for operating systems and
processes in almost all industrial plants. Moreover, the carbon footprint of a water
supply system in a plant has wide impacts on those of the various products produced
in the plant. Besides that the application of reuse water for landscape plants may
also reduce carbon footprint where leave grass clippings on the yard can decompose
and return nutrients to the soil. Planting trees, vines, bushes, mushrooms, and
ground-covers can also reduce carbon footprint (Lim et al., 2007).
2.10
Carbon Footprint
Carbon footprint is a frequently used term that has no precise scientific
definition for fossil CO2 emissions.
A commonly accepted description is "The
carbon footprint is a measure of the exclusive total amount of carbon dioxide
42
emissions that is directly and indirectly caused by an activity or is accumulated over
the life stages of a product." (ISA Research Report, UK 2007). The term embedded
carbon emissions is also used for products.
European carbon emissions continue to rise by over 1% per year, in spite of
the Kyoto treaty’s commitment to reduce them to the 1990 level by 2012. The
announced 2020 European target is a 20% reduction compared to 1990 and 75% by
2050. It is estimated that an average person in the UK contributes about 11 tonnes of
carbon to the atmosphere each year through their non-professional actions.
Politicians are increasingly looking at standards, labels and other instruments
relevant to consumers to involve them in climate change mitigation. Therefore,
attention goes beyond carbon emissions of production activities, companies or
sectors, and is also focusing on emissions associated with products. The principal
actions to reduce the carbon footprint are:
a. Implement an environmental and energy management strategy.
b. Measure energy consumption (from fossil fuels) and identify areas for
improvement.
c. Reduce energy consumption by improving efficiency of production
technologies, their use and maintenance. This includes secondary use
of hot air and water, and power generation.
d. Improve the energy efficiency of buildings and their services.
e. Reduce transportation energy used
43
2.11
Water Reuse Problem
Recent studies support long standing concerns about possible public health
effects of reclaimed water. It has been known for some time that treated waste water
effluent, or reclaimed water, contains pathogens that could be transferred to people
through contact, including aerosols from sprinklers.
However, disease causing
agents such as E.coli, Salmonella, Giardia intestinalis, Entamoeba histolytica and
viruses are known to be present in effluents and might cause a disease outbreak if
exposed to public (Rose et al., 1996).
Particularly worrisome are high levels of
parasites such as Giardia and Cryptosporidium which are not killed by chlorination.
In 1997, the EPA warned, “(Viable) bacteria from reclaimed water in
sprinklers can travel more than 1000 feet in the air”.
As far back as 1984,
researchers concluded that disinfection by chlorination, an important part of
wastewater treatment, initially lowered the total number of sewage related bacteria,
but may substantially increase the proportions of antibiotic resistant, potentially
pathogenic organisms used (Hammer et al., 2008). More recently, Chang (2007)
reported that Staphylococcus aureus bacteria become more virulent and drug
resistant after chlorination. A large study in 2006 confirms that microbes, inactivated
but not killed by treatment, can regrow in retention ponds and pipes, becoming a
major source of the spread of multi drug resistant pathogens in the environment.
The principal mechanism for overcoming any difficulties relating to reusing
water for irrigation of crops is the pre-treatment of the recycled water. As mentioned
above the risk from microbial pathogens is significantly reduced with the treatment
of water (Peasey et al., 2000).
Treatment of recycled water also reduces the
concentration of organic and inorganic nutrients, trace organics and heavy metals.
The major contaminant that is difficult to remove from recycled water is salt and
other cations and anions. The only effective treatment mechanism to remove salt
molecules and ions is reverse osmosis membrane filtration. Such a high level of
44
treatment is far too expensive to be economically viable for irrigation of crops and
turf.
2.12
Wastewater Treatment
With the increasing urbanization and industrialization, the activities of
mankind give rise to a wide range of waste products, many of which become
waterborne and must be carefully treated before being released to the environment.
Meanwhile, the volume of domestic sewage, industrial effluents, agricultural wastes,
and urban runoffs, is steadily growing. Wastewater is one of the most important
problems occurring and requires proper treatment before discharge to receiving
watercourses. Wastewater needs to be adequately treated prior to disposal or reuse in
order to (Mara, et al, 1992):
a) protect receiving water from contamination as the receiving water is
generally used as a source of untreated drinking water by downstream
communities (or, in the case of coastal waters, used for shellfisheries);
b) protect receiving waters from deleterious oxygen depletion and ecological
damage; and
c) produce microbiologically safe effluents for agricultural and aquacultural
reuse (for example, crop irrigation and fishpond fertilization).
There is an increasing need for low-cost methods of treating wastewaters,
particularly municipal sewage and industrial effluents. The operation of such
methods, and the maintenance of the necessary plant and equipment, must be within
the capability of developing urban centers and industrial complexes. It is generally 6
45
recognized that some form of biological treatment provides the most economical
solution for handling domestic and most industrial wastewater.
2.13
Wastewater Treatment in Malaysia
2.13.1 Environmental Regulations and Standards in Malaysia
Since independence, Malaysia has forged ahead with rapid economic growth.
The implementation of various development plans has resulted in the setting up of
manufacturing industries and intensification of agricultural activities to provide
employment and export earnings. However, Malaysia’s rapid economic development
has drawn heavily on its resource base ranging from forestry, land and water to fossil
fuels and minerals. Air, noise and water pollution and industrial waste generation and
degradation of the environment are of serious concern arising from such
development.
Fully aware of the environmental problems and the consequences that would
follow if no actions were taken; the Environmental Quality Act (EQA) 1974 was
enacted by the Government of Malaysia after an inter-government committee study.
One of the three strategies embodied in the EQA 1974 is to regulate pollution. The
underlying principles adopted in the formulation of EQA are (Rahani, 1995):
a) pollution should be controlled at source;
b) polluters must pay or bear the costs of their waste or wastewater treatment
or disposal;
c) discharge standards should be uniform for a particular source, type of
industry or activity;
46
d) uniform discharge standards could be contravened for polluters whose
discharge does not adversely affect receiving waters, i.e. waters still have the
required “carrying capacity”; so-called “polluters” must bear the costs to
investigate studies required by the relevant authorities; and
e) variable discharge standards ought to be introduced by the Minister of the
Environment should the uniform standards imposed on every discharge point
and all polluting sources within a water body are inadequate to maintain the
conditions necessary to support the intended use of the water body.
2.14
Wastewater Treatment Method
Wastewater can be defined as water that has been used and it could be
polluted (Department of Environmental Engineering, 2006).
The needs are
important to treat wastewater because it can be harmful to health and affect the
ecosystem. There are many types of wastewater treatment that is being used in
Malaysia such as coagulation, disinfections, advanced wastewater treatment and etc.
There are also different advantages and disadvantages for different types of treatment
that have been used. Coagulants may be added during treating wastewater due to its
ability to destabalise particles and enable them to become attached to the other
particles. So, it is easier to remove particles. The most common used coagulants in
water and wastewater treatment currently are aluminium sulfate (alum), ferric
chloride, ozonation and ultraviolet (UV).
47
2.14.1 Aluminium Sulphate (Alum)
Aluminium sulphate (alum) is a compound derived from aluminium, one of
the earth’s most abundant metals. Aluminium has a high ionic charge and a small
crystalline radius which combine to yield a level of reactivity which is unmatched by
any other soluble metal. Alum has been used in water purification and wastewater
treatments for centuries and lake restoration for decades. Moreover, it is harmless to
aquatic plants and water creatures (Harper, 2007).
The advantages of alum are well-known in water treatment. First, alum
coagulation provides rapid, highly efficient removal of solids, phosphorus and
bacteria. Liquid alum is relatively inexpensively, resulting in low unit costs per mass
of pollutant removed. Unlike iron compounds, alum do not deteriorate under longterm storage. Due to the quality of raw materials used for manufacture of alum,
liquid alum contains substantially less heavy metal contamination than other metal
coagulants. Alum floc is chemically inert and is immune to dissolution from normal
fluctuations in pH (Harper, 2007).
However, as a conventional coagulant, aluminium has long been suspected of
being both carcinogenic and mutagenic.
Given the possible health risks, it is
necessary to monitor aluminium residue in treated water and take appropriate
measures to keep it below a certain level.
Besides, there are additional costs
associated with floc disposal and maintenance (Selvarajoo, 2006). Another problem
is reaction alum with alkalinity in water to decrease the pH value in water
coagulation also not very effective (Katayon et. al., 2006). Sulphate salt and syntetic
polymer are the one alternative that used in coagulation but the reagents that been
used are costly and some developed country cannot afford the cost of chemical
reagents for the water and wastewater method (Diaz et. al., 1999; Schulz and Okun,
1984).
48
2.14.2 Ferric Chloride
Ferric chloride has been widely used in water treatment due to its good
performance on turbidity removal.
It is more effective than aluminium-based
coagulants in removing total organic carbon (TOC), achieving the same removal
effect with a lower dosage. Ferric chloride coagulants are also found to be effective
in wide ranges of pH and temperature in removing humic substances from water, a
main trihalomethanes (THMs) precursor when coagulated water was chlorinated.
Beside that, iron-based coagulants do not pose the health risks of their aluminium
counterparts.
2.14.3 Ozonation
Ozone is a strong oxidizing substance with bactericidal properties similar to
those of chlorine. Test conditions show that the destruction of bacteria is between
600 to 3000 times more rapid by ozone than by chlorine. Further, the bactericidal
action of ozone is relatively unaffected by changes in pH while chlorine efficiency is
strongly dependent on the pH of the water (Chen and Nicholas, 2007). Besides the
disinfections of sewage effluent, ozone is used for sterilizing industrial containers
such as plastic bottles where heat treatment is inappropriate. Breweries use ozone as
an antiseptic in destroying pathogenic ferments without affecting the yeast. It is also
used in swimming pools and aquariums.
Ozonation of water is a well-known technology and the strong oxidative
properties of ozone have been well documented (Lin et al., 2001). Ozone treatment
of several types of wastewaters, resulting in considerable COD reduction has been
reported by several researchers but ozonation process may release more toxic
products in treated wastewater. As a strong oxidant, ozone reacts with a wide variety
49
of organics.
Ozone oxidizes phenol to oxalic and acetic acids. It oxidizes
trihalomethane (THMs) compounds to a limited extent within proper pH ranges and
reduces their concentration by air stripping. Oxidation by ozone does not result in
the formation of THMs as does chlorination. Ozonation treatment are most costly
than chlorination but it has more advantages to colour abolishment (Ruslan Hassan,
1988).
2.14.4 Ultraviolet (UV)
The properties of ultraviolet (UV) radiation sources have been used in a wide
variety of applications since the use of UV was pioneered in the early 1900s, having
been discovered first in the 1880s. With the proper dosage, UV irradiation has
proven to be an effective disinfectant for bacteria, protozoa and virus in reclaimed
water, while not contributing to the formation of toxic bioproducts (Metcalf and
Eddy, 2004).
The portion of the electromagnetic spectrum in which UV radiation occurs,
is between 100 and 400 nm. The effectiveness of the UV disinfection process
depends on the characteristics of the microorganisms. Typical values for the
disinfection of coliform organisms with UV light for various wastewater (Metcalf
and Eddy, 2004). One of the most serious problems encountered with UV
disinfection systems in open channels is achieving a uniform velocity field in the
approach and exit channel.
Although, in general, it is believed that the concentration of suspended solids
has a deleterious impact on UV disinfection performance. The advantages of using
UV are there are no odor and taste effects, minimum of surveillance; ease automatic
50
guard without dangers because of over dose. The advantages of using the ultraviolet
is very costly than other treatment (Ruslan Hassan, 1988).
2.14.5 Disinfection
Disinfection normally involves the injection of a chlorine solution at the head
end of a chlorine contact basin. The chlorine dosage depends upon the strength of
the wastewater and other factors, but dosages of 5 to 15 mg/l are common. Ozone
and ultra violet (UV) irradiation can also be used for disinfection but these methods
of disinfection are not in common use (Metcalf and Eddy, 2004). Chlorine contact
basins are usually rectangular channels, with baffles to prevent short-circuiting,
designed to provide a contact time of about 30 minutes. However, to meet advanced
wastewater treatment requirements, a chlorine contact time of as long as 120 minutes
is sometimes required for specific irrigation uses of reclaimed wastewater. The
bactericidal effects of chlorine and other disinfectants are dependent upon pH,
contact time, organic content, and effluent temperature.
The purpose of “disinfection” in the treatment of wastewater is to
substantially reduce the number of microorganisms in the water to be discharged
back into the environment. The effectiveness of disinfection depends on the quality
of the water being treated (e.g. cloudiness, pH, etc.), the type of disinfection being
used, the disinfectant dosage (concentration and time), and other environmental
variables (Hammer et al., 2008).
Disinfection is the destruction or inactivation of harmful microorganism in
water by either a physical process or a chemical process. Physical disinfection such
as heating is safe and would destroy pathogenic microorganisms such as viruses,
51
bacteria, and so on. But it is not applicable on large scale, as it is expensive (Kumar
et al., 2006). Chemical disinfection is the most widely used method. The chemicals
used are chlorine and chlorinated compounds, bromine, iodine, ozone, potassium
permanganate, hydrogen peroxide, silver and even some phenolic compounds.
Chlorine and its various compounds are most widely used for disinfection.
They are used because of the several advantages such as ease of destruction of
pathogens, cheapness and wide availability. It also eases of application with high
solubility in water and non-coloration of the treated water. Since chlorine is a
poisonous gas, it requires careful handling. It may form carcinogenic, taste and odor
forming compounds when it reacts with some organic compounds (Kumar et al.,
2006).
2.15
Previous Study
Previous study on effluent reuse on landscape plants uses chlorine for
disinfection purposes (Mohd Ismid et al., 2003). Chlorine was used due to several
advantages such as ease of destruction of pathogens, inexpensive and wide
availability. It also eases of application with high solubility in water and noncoloration of the treated water. Since chlorine is a poisonous gas, it requires careful
handling. It may form carcinogenic, taste and odor forming compounds when it
reacts with some organic compounds (Schulz et al., 1984).
There is also the study have been conducted on reuse of wastewater for food
crops by using chlorination for disinfection purpose. The study was carried out to
examine the suitability and feasibility of wastewater reuses in concern of human
health and the environment and look forward the efficiency of wastewater reuses of
crops growth (Rubiah, 2005). Two types of crops being studied which is chili
52
(Capsicum annum) and brinjal (Solanum melongena). From the previous study, it can
be conclude that wastewater reuses can also be considered as a potential resource
after the proper treatment process and disinfection (Rubiah, 2005).
The purpose of disinfection in the treatment of wastewater is to substantially
reduce the number of microorganisms in the water to be discharged back into the
environment. The effectiveness of disinfection depends on the quality of the water
being treated (e.g. cloudiness, pH, etc.), the type of disinfection being used, the
disinfectant dosage (concentration and time), and other environmental variables
(Hammer et al., 2008). In this study, Terminalia catappa leaves will be applied in
the wastewater treatment to replace chlorination.
Another previous study also has been conducted to understand the effect of
wastewater effluent on cultivation of chili plant, Capsicum annum by using
hydroponics drip irrigation system (Parameswary, 2008). The study was observed
the capability of the chili crops Capsicum annum to grow productively in three
batches of samples receiving wastewater, fertilizer or tap water. The plants grown
using wastewater effluent under drip irrigation obtained the highest leaves width and
plant heights compared to that of fertilizer and tap water plants.
2.16
Terminalia catappa
The main material that is used in the study is Terminalia catappa leaves in
order for replacing chlorination process to kill bacteria. Terminalia catappa is a
species of tropical tree that grows in Asia. The tree's origin is controversial, and
could have been India, Malay Peninsula, or New Guinea. With so many common
name its have, it just comes from COMBRETACEAE (Combretum Family) (Gilman
et al., 1994). Common names include Indian Almond, Bengal Almond, Singapore
53
Almond, Malabar Almond, Tropical Almond, Sea Almond, and Umbrella Tree. It is
also known as Java Almond in Indonesia, Tropical Almond in India, Hwu Kwang in
China, and Kobateishi in Japan. There are also name like False Kamani; Almendras
and Myrobalan (Ida Murni, 2008).
2.16.1 Characteristics of Terminalia catappa
The Terminalia catappa tree grows to 35 m height, with an upright,
symmetrical crown and horizontal branches.
As the tree gets older, its crown
becomes more flattened to form a spreading, vase shape (Figure 2.3). The leaves are
large, 15-25 cm length and 10-14 cm width, ovoid, glossy dark green and leathery
(Figure 2.4).
They are dry-season deciduous; before falling, they turn pinkish-
reddish or yellow-brown, due to pigments such as violaxanthin, lutein, and
zeaxanthin (Figure 2.5 & Figure 2.6).
Figure 2.3: Terminalia catappa tree
Figure 2.4: Illustration of Terminalia
catappa Leaves
54
Figure 2.5: Terminalia catappa leaves
Figure 2.6: Terminalia catappa Leaves
Before Falling
After Falling
The flowers are monoecious, with distinct male and female flowers (Figure
2.7) on the same tree (Chen and Nicholas, 2007). Both are 1 cm diameter, white to
greenish, inconspicuous with no petals; they are produced on auxillary or terminal
spikes. The fruit (Figure 2.8) is a drupe 5-7 cm long and 3-5.5 cm broad, green at
first, then yellow and finally red when ripe, containing a single seed and does not
attract wildlife, inconspicuous and not showy.
Figure 2.7: Terminalia catappa Flowers
Figure 2.8: Terminalia catappa Fruits
55
Terminalia catappa is a large deciduous stately tree, originally from India,
growing up to 90 feet tall with horizontal whorls of branches offering clusters of foot
long. The trees are availability grown in small quantities by a small number of
nurseries. The trunk or branches droop as the tree grows, and will require pruning
for vehicular or pedestrian clearance beneath the canopy; routinely grown with, or
trainable to be grown with, multiple trunks; not particularly showy; tree can grow
with several trunks but can also be trained to grow with a single trunk and no thorns.
The tree may be best suited for planting along the coast as a park or shade
tree providing dense shade. People may object to the large leaves and the fruit that
falls from the tree if the tree is used as a street tree, and the tannic acid may be a
problem near parked cars. Branches droop and require regular maintenance to keep
them pruned to allow for vehicle clearance beneath the canopy. However, it would
make a nice tree for a median or along a boulevard where this would cause fewer
nuisances (Gilman et al., 1994). Terminalia catappa should be grown under bright
sunlight on any well-drained soil. The plant is quite tolerant of wind, salt, and
drought but need protection from freezing temperatures.
Trees perform best if
mulched and regularly fertilized.
2.16.2 The Uses of Terminalia catappa
Terminalia catappa is a Combretaceous plant (Tropical Almond family),
presenting throughout any region of Thailand. The plant is a large tree, which can
reach up to 30 m height with a thick broad trunk; the leaves cluster toward the end of
the branches with glossy, obovate blades mostly 8 to 30 cm in length and turn red
before turning brown and falling. The leaves take an antiseptic effect. They are used
to treat fungus and bacteria disease.
56
Besides creating a natural environment and induces spawning, it also controls
or reduces the pH level of water, thus it is also one of the best aquarium water
conditioner. They are known to have antibacterial, antifungal, stresses relieve and
immune boosting properties for all tropical fishes (Grandiosa, 2002). In the study,
the Terminalia catappa leaves can be applied in the wastewater treatment to replace
the “chlorination” process.
In aquaculture, Terminalia catappa leaves have been claimed as a promoting
substance for wound healing, especially for injured Siamese fighting fish after
fighting matches. Chansue et al., (2002) reported that it increase thickness of keratin
layer in Siamese fighting fish scale. The leaves have a potential to use as an
alternative treatment for chemical substances and antibiotics. Various concentrations
of the extracts in water to prevent fish pathogen have been examined. Chansue and
Tangtrongpiros (2005) found that water extracts of the dry leaves can rapidly
promote regeneration of fin tail of fancy carp.
Chitmanat et al. (2005) reported effectiveness of 0.8 mg/l concentration of
leaf extracts of Indian almond tree against Trichodina and other bacterial infections
in tilapia, and against fungal infection in tilapia egg. In addition, the leaf extracts can
eliminate Zoothamnium spp. infection of black tiger postlarva shrimp within 24
hours after exposure, and can significantly decrease the number of Gyrodactylus and
Dactylogyrus infection of gold fish.
In Thailand, leaves of Terminalia catappa tree have been widely used in
Siamese fighting fish culture as bath supplement for treating the injured fish and
promoting the fish breeding. However, the breeders still use their experiences to
estimate the concentration of the leaves. Chansue, (2005) reported effects of the
extract on hematology and blood chemistry of Siamese fighting fish but did not
report on its toxicity level.
57
Terminalia catappa has strong antibacterial properties and works against
Gram positive and Gram negative micro-organisms. The leaves have been shown to
protect against acute liver injury produced by some hepatotoxicants (chemicals that
produce liver damage). In Nigeria fallen leaves are used as herb to treat liver
diseases. The leaves also have potential in the management of sickle cell disorders.
Dried leaves are used for fish pathogen treatment, as an alternative to the use of
chemicals and antibiotics. Leaves have antioxidant as well as anti-clastogenic
(preventing breakage of chromosomes) properties. The various extracts of leaves and
bark of the plant have also been reported to be anticancer, antioxidant, anti-Human
Immuno-deficiency Virus (HIV), reverse transcriptase and anti inflammatory (Chua
et al., 2007).
In Taiwan the fallen leaves of tropical almond are used as an herbal drug in
the treatment of liver related diseases.
The leaves contain agents for chemo-
prevention of cancer and probably have anti-carcinogenic potential. They also have
an anticlastogenic effect (a process which causes breaks in chromosomes) due to
their antioxidant properties. The kernel of Indian almond has shown aphrodisiac
activity; it can probably be used in treatment of some forms of sexual inadequacies.
The extract of Terminalia catappa leaves has been tested and the result show
antibacterial activities occurrence (Suganda et al., 2004). According to Chansue,
(2008) the results show the total tannin level increased when duration of extraction
was increased. Liu et al. (1996) reported that the water extract of the dried leaves of
Terminalia catappa inhibited lipid peroxidation in vitro and TPA-induced hydrogen
peroxide formation in human mononuclear leukocytes, depending on the dosage
used. It was reported that the anticlastogenic effect of Terminalia catappa leaves
might be attributed to their antioxidant potential (Liu et al., 1996).
In addition, Chen et al. (2000) found that the water extract from fallen leaves
and punicalagin were effective against bleomycin induced genotoxicity in Chinese
hamster ovary cells. Chen et al. (2000) also indicated that the effectiveness could be,
58
at least in part, due to their antioxidant potential. Although many synthetic
chemicals, such as phenolic compounds are found to be strong radical scavengers,
they usually have side effects (Imaida et al., 1983). Antioxidant substances obtained
from natural sources will be of great interest. The high contents of tannins in
Terminalia catappa leaves reveal that they may serve as a source of natural
antioxidants.
2.16.3 Tannin
Tannins are distributed all over the plant kingdom. They are commonly
found in both gymnosperms as well as angiosperms. In terms of location of the
tannins in a plant, they are mainly located in the vacuoles or surface wax of the
plants. These sites are where tannins do not interfere with plant metabolism, and it is
only after cell breakdown and death that the tannins are active in metabolic effects
(Corder et al., 2006). Tannins are found in leaf tissues, bud tissues, seed tissues, root
tissues and stem tissues. An example of the location of the tannins in the stem tissue
is that they are often found in the growth areas of trees, such as the secondary
phylum and xylem and the layer between the cortex and epidermis.
Tannins may help regulate the growth of these tissues. They are also found in
the heartwood of conifers and may play a role in inhibiting microbial activity, thus
resulting in the natural durability of the wood. However, there may be a loss in the
bioavailability of tannins in plants due to birds, pests, and other pathogens (Corder et
al., 2006). The leaching of tannins from the decaying leaves of vegetation adjoining
a stream may produce what is known as a black water river
Tannins, lignins and fulvic acids are sub classes of humic acids. They all tint
the water yellow. The freshwater humic acids can come from a variety of sources,
59
most of which are on land (decomposing terrestrial vegetation.) These substances
wash into lakes and rivers, undergoing further transformations along the way, and
ultimately into the ocean. Humic acid contains sulfur, nitrogen and phosphorus in
varying amounts. It also contains metals such as Ca, Mg, Cu and Zn which can be
'chelated' in some undefined way (Chansue, 2008).
Humic acid can be broken down into two groups based on the polarity and
size of the individual compounds. The smaller, more polar fraction is generally
termed fulvic acid and the larger; more non-polar fraction is generally termed humic
acid. Humic acids are the end product of microbial degradation of plant and animal
debris and are one of the most important constituents of fertile soils. Tannic and
humic acids may be useful for inhibiting many types of bacteria including cyanobacteria and are fairly benign for fish (Chansue, 2008).
According to previous studies, the total tannin level was rapidly increased
within the first three days of extraction then gradually increased. We can get higher
concentration of tannin when allowing longer time of extraction (Chansue, 2008).
However, the longer the time, the higher the number of microorganisms
contaminated. Therefore, the extract of Indian almond leaves for 3 days was the
most appropriate for use. Chitmanat et al. (2005) also reported that water solution of
dried Terminalia catappa leaves (0.8 ppm) was able to inhibit A. hydrophila
infection in tilapia. The extracts are gradually acidic when their concentrations
increase.
2.17
Escherichia coli
The bacterial species Escherichia coli (E. coli) is one of the most common
inhabitants of the human intestinal tract and is probably the most familiar organism
60
in microbiology. Its presence in water or food is an indication of fecal contamination.
E. coli is not usually pathogenic (Gerard et al., 2004) as shown in Figure 2.9.
However, it can cause urinary tract infections, and certain strains produce
enterotoxins that cause traveler’s diarrhea and occasionally cause very serious
foodborne disease.
E. coli are Gram-negative bacteria that belong to the g-
proteobacteria. As they primarily live in the mammalian gut they have been grouped
with other related bacteria as 'enteric' bacteria. They are straight rod shaped cells of
about 2 µm long and 0.5 µm wide, which can grow and divide rapidly by binary
fission.
There are many different types of E. coli and the chief way they are
distinguished is immunologically using sterotyping.
Figure 2.9: Escherichia coli (E. coli) (Gerard et al., 2004)
2.18
NPK Ratio
A proper understanding of nitrogen-phosphorus-potassium (NPK) ratio will
ensure that crops are properly fed and nourished. These essential nutrients promote
vigorous growth, increased root development and improved disease and stress
tolerance. According to Sustainable Gardening Australia (2009), these three
61
macronutrients are the most popular nutritional elements required by plants, as they
are all needed in the largest volumes for healthy plant growth. Our top three are
nitrogen, phosphorus and potassium.
a)
Nitrogen (N)
Nitrogen is a vitally important element in the plant amino acids (the building blocks
of proteins), nitrogen is also a necessary part of the green bits in the leaves of plants
(chlorophyll) and super dooper for leaf growth. Plants may be nitrogen deficient if:
older leaves are pale green to yellow often with a tinge of red, plants are stunted, leaf
drop can occur. Plants may have too much nitrogen if: shoots wilt; dead spots appear
on the leaves of young plants; masses of soft deep green leaves that are prone to pest
and
b)
disease
appear
(Sustainable
Gardening
Australia,
2009).
Phosphorus (P)
Phosphorus is involved in just about everything that goes on in plants. It is especially
important where cells are actively dividing (growing), and it promotes the growth of
seedlings, roots, flowering, and fruit and seed formation. Plants may be phosphorus
deficient if: they have poorly developed roots, plants are a bit stunted, and, most
obviously, leaves have a purple tinge to them (Sustainable Gardening Australia,
2009).
Plants may have too much phosphorus if: older leaves display necrosis (browning
and death) of tips and margins followed by dropping off of the affected leaves.
Chlorosis (yellowing) of younger foliage often occurs and the plant dies in severe
cases. Phosphorus toxicity is rare in many species but there a small number of
species that are most prone to the problem. Of these species the majority are
Australian natives from the PROTEACEAE family.
62
c)
Potassium (K)
Potassium is extremely important in helping plants resist disease, producing
chlorophyll, making plant cells stronger and moving water through plants. Plants
may be potassium deficient if: they have weak stems, older leaves are floppy and
have brown scorched edges and can be bluish green, and may be overly sensitive to
frost. If you live in an area with really sandy soils and a fair bit of rainfall, you may
just come across this deficiency. Plants may have too little potassium if: they display
similar symptoms to phosphorus deficiency, but with the added bonus of poor root
development and wilting of new growth (Sustainable Gardening Australia, 2009).
2.18.1 The Secondary Macronutrients
Like their three big brothers the primary macronutrients, secondary
macronutrients are also essential for healthy plant growth, but are needed in smaller
quantities for success. These three champions are calcium, magnesium and sulphur.
a)
Calcium (Ca)
Plants may have a calcium deficiency if: you have highly acid soils and notice weak
plants stem, and see stunting or dead spots on the newest leaves. Plant roots will
often be stunted, brown and dead. Leaves of grasses and grass-like plants may fail to
form properly, and appear rolled. Calcium is often present in soils in correct
quantities, but, due to other factors (pH, other nutrient deficiencies); plants are not
63
able to "access" and use this micronutrient. Plants may have too much calcium if:
they display signs of magnesium and boron deficiency. Rarely do plants have too
much calcium, but, in highly acid soils, excessive calcium take up can make
magnesium and boron unavailable to the plant (Sustainable Gardening Australia,
2009).
b)
Magnesium (Mg)
Plants may be magnesium deficient if: older leaves of the plant turn yellow while the
veins remain green (this cool looking symptom is commonly referred to as
"intervienal chlorosis" and is generally caused by an iron deficiency). Some plants,
especially grasses, may have the edges of the leaves turned upwards, and, if the
deficiency is severe, they may turn a reddish-brown colour. Plants may have too
much magnesium if: well, it's unlikely this would happen, but magnesium toxicity
would present first as a potassium deficiency (Sustainable Gardening Australia,
2009).
c)
Sulphur (S)
Sulphur is responsible for many of the flavors and odors of plants - think cabbage,
Brussels sprouts and asparagus. Sulphur is also incredibly important in the
production of chlorophyll and many plant proteins. Plants may be deficient in
sulphur if newer leaves turn yellow, and new shoots are stunted. If left for ages, these
symptoms will appear very similar to those of nitrogen deficiency, the difference
being whether the yellowing appeared first in the new leaves (sulphur) or the older
leaves (nitrogen). Plants may have too much sulphur if leaf size is reduced and
overall growth stunted with newer leaves yellowing or scorched at edges. Older
leaves may drop prematurely (Sustainable Gardening Australia, 2009).
CHAPTER 3
METHODOLOGY
3.1
Introduction
Figure 3.1 and 3.2 shows the design framework for this study where it
illustrates the overall process and experiment that will be conducted and also their
flow of work. At the initial stage, the leaves were collected and only brown coloured
leaves were selected in the study. The Terminalia catappa leaves were collected
from trees located near Padang Kawad, Universiti Teknologi Malaysia (UTM). The
leaves must be in clean and dry condition.
65
Part 1
Collect Terminalia catappa
leaves at Padang Kawad, UTM
Clean and Dry the leaves
Extract tannin for Terminalia
catappa leaves
•
•
Jar Test
To obtain optimum dosage
Different condition of leaves
(Blended with water, original form and cut form)
Soxhlet Extraction for
comparison purpose were
done in Organic Research
Laboratory, Faculty of
Science (UTM)
Wastwater Sampling at Oxidation Pond near Kolej Tuanku
Canselor (KTC), UTM
Treatment of
Terminalia catappa
Leaves with the
optimum dosage
After 5 days, transfer to
storage tank at
landscape garden
Applied 2
times a day
Coliform
analysis
Wastewater analysis
•
Biological oxygen demand
•
Chemical oxygen demand
•
pH
•
Ammonical nitrogen
•
Tannin
•
Phosphorus
•
E.coli
Figure 3.1: Design Framework of Part One for this study
Chlorination
process for
comparison
purpose in
order to kill
bacteria
Iodometric
Method to
determine the
residual of
chlorine
66
Part 2
Design of two Landscape
Plots
An open-air landscape
garden of 1.5 m x 1.0 m
x 0.3 m was constructed
Five differents shrubs and
one types of grass were
planted in the garden
Coleus
artopurpureus
(Ca)
Cyperus
dubius
Rottb
(Cd)
•
Two differents
storage tanks were
applied in the garden
•
Water from storage
tank were applied to
garden every morning
and evening
Nyctanthes
arbo-tristis
L (Na
•
•
Panicaum
maximum
cv. Colonio
(Pm)
•
•
Vinca
rosea
(Vr)
Determination of
NPK Ratio from
the soil
pH value of the soil
Elephantopus
Scaber (Es)
Microbiology Sampling
Procedures for Leaves and Soil
(E. coli)
Conducted for three times a
week
Shrubs Growth
Determination
Shrubs Height
Leaves Width
Figure 3.2: Design Framework of Part Two for this study
67
3.2
Jar Test
Jar test experiment was used to determine the most effective and economical
dosage of catappa leaves in order to kill bacteria. In this study, Jar test (Figure 3.3)
was used to determine the optimum dosage of Terminalia catappa (TC) leaves in
treating wastewater.
Figure 3.3: Jar Test Analysis
3.3
Leaf Condition Analysis
The analysis was conducted to determine the best leaf’s condition that was
used in this study and consequently to prove that the Terminalia catappa leaves
extract is the best form. The alternatives for leaf conditions are blended leaves,
sliced leaves and original form of leaves. In this experiment, five beakers were
soaked with catappa leaves with different form of leaves. Each beaker contains
different dosage of catappa leaves i.e. 30 g, 40 g, 50 g, 60 g and 70 g. For this
experimental setup, the dried leaves will be minced and soaked in 1 litre of distilled
water for 7 days at room temperature. The extracted solution will then be filtered
68
and the total tannin concentration released will be measured daily using HACH
DR5000 Spectrophotometer.
pH measurement will be also conducted daily to
determine the best state of catappa leaves in order to eliminate bacteria.
3.3.1
Optimum Dosage of Terminalia catappa Leaves (Tannin Concentration)
In order to ascertain tannin concentration in TC leaves, a series of jar test
analysis was conducted to measure the optimum dosage of Terminalia catappa
leaves needed in order to kill bacteria or pathogens. Leaves were collected in dried
form i.e. brown in colour. Each beaker contains different dosing of catappa leaves
which are 30 g, 40 g, 50 g, 60 g and also 70 g as shown in Figure 3.4. The dried
leaves were minced and soaked in 1 liter of distilled water for 7 days at room
temperature.
The extracted solution was then filtered and the total tannin
concentration was determined using HACH DR5000 Spectrophotometer and the pH
measurement was also conducted.
Then, from the optimum dosage calculated,
catappa leaves were soaked in 1 liter of water sample for 1, 3, 5 and 7 days at room
temperature. The tannin solution was then applied to the sewage effluent to evaluate
its antibacterial properties using E. coli analysis.
Figure 3.4 Jar Test analysis for optimum dosage of tannin concentration
69
3.4
Extraction of Terminalia catappa Leaves
Extraction of TC leaves using Soxhlet extraction was also carried out for
comparison purposes. The Soxhlet experiment (Figure 3.5) was conducted in
Organic Research Laboratory, Faculty of Science, UTM. The dried leaves were
subjected to size reduction to small size and 50 g each of the dried leaves were used
for the extraction. For the preparation method, the Soxhlet extraction thimble was
dried at 105°C to constant weight. Then, 50 g of Terminalia catappa leaves were
packed in a thimble of Soxhlet apparatus. Making sure that the thimble was not
overfilled and leaving at least 1 cm gap between the sample and the top of the
thimble.
Figure 3.5: Example of Soxhlet Extraction carried out at Organic Research
Laboratory, Faculty of Science (UTM)
Then, the leaves were extracted with 200 ml of solvent (in the ratio of 9:1ml
methanol: distilled water respectively). The leaves were completely submerged and
then covered with Glass wool. Extraction (Figure 3.6) was allowed to proceed for 24
hours. The extract was then decanted and the solvent removed by evaporation at
70
room temperature (28 + 2oC) to obtain the extract. The air dried extract was stored
for 24 hours in sterile universal bottles at room temperature. The sterility of the
extract was tested before use. Another extraction was also carried out for 70g leaves
to compare the tannin concentration in each sample.
Figure 3.6: Example of Soxhlet Extractor with Terminalia catappa Leaves
3.5
Wastewater Collection
Treated sewage effluent sample will collected from oxidation pond near
Kolej Tuanku Canselor (KTC) at Universiti Teknologi Malaysia (UTM) Skudai,
Johor before it was discharged to the river.
This treatment plant is the major
treatment facility at Universiti Teknologi Malaysia (UTM), which processes an
average of 250,000 m3/day wastewater originating from the campus and staff
residential area. The treatment primarily involved oxidation process with a retention
time of 24 hours. For this study, effluent was collected weekly in the morning.
Figure 3.7 shows that the oxidation pond where the sample was collected.
71
Figure 3.7: Oxidation pond near Kolej Tunku Canselor, UTM
3.6
Sample Handling Procedure
After sample is taken from oxidation pond and brought to the laboratory,
immediate analysis was conducted on pH, BOD and E. coli. All these parameters are
sensitive and their value will differ even after only one day. The remainder of the
sample was then kept refrigerated at 4°C. The samples were then reduced to room
temperature for analysis.
3.7
Apparatus and Chemicals
List of apparatus and chemicals that used during this study are simplified and
tabulated in Table 3.1.
72
Table 3.1: List of apparatus and chemicals for each test
Water Parameter
Instrument
Program No. for HACH
DR5000 Spechtophotometer
pH
pH meter Thermo Orion
-
Model 420 A
Biological oxygen
BODTrak apparatus
-
Chemical oxygen
HACH DRB200
2730
demand (COD)
HACH DR5000
demand (BOD)
Spechtophotometer
Ammoniacal Nitrogen
HACH DR5000
1700
Spechtophotometer
Phosphorus
HACH DR5000
3850
Spechtophotometer
Tannin
HACH DR5000
3550
Spechtophotometer
Escherichia coli
Quanty-Tray/2000
Quanti-Tray Sealer
Model 2x Thermolyne
Type 142300 Incubator
-
73
3.8
Wastewater Treatment using Terminalia catappa Leaves
After the Jar Test has been conducted, the result of optimum dosage of
Terminalia catappa leaves will be obtained. The treatment of domestic effluent
using the optimum dosage was used where the catappa leaves was minced and
soaked in the tank as shown in Figure 3.8. In this treatment, catappa leaves was
soaked in the tank with 25 liters of water sample for five days. From the previous
study, the results show that the total tannin level increased when duration of
extraction was increased. The wastewater was then analyzed for pH, COD, BOD,
tannin,
ammonical
nitrogen
and
phosphorus
using
HACH
DR5000
Spectrophotometer. For E. coli test, it was used colilert to measured the number of
bacteria contains in the wastewater. After that, the effluent was transferred to another
storage tank and applied everyday at a volume of 5 liters.
Figure 3.8: Treatment of wastewater using Terminalia catappa Leaves in the tank
3.9
Water Quality Analysis
By using the optimum dosage of Terminalia catappa leaves and the best leaf
condition, the leaves was explored to investigate its antibacterial properties in order
74
to eliminate bacteria in sewage wastewater.
Then, the outcome was tested for
several water parameters such as pH, biochemical oxygen demand (BOD), chemical
oxygen demand (COD), ammoniacal nitrogen (AN), phosphorus, tannin and E. coli.
The main objectives are to identify the antibacterial properties of Terminalia catappa
leaves and to compare the state of Terminalia catappa leaves with chlorination
process to kill bacteria. The result obtained from Terminalia catappa leaves and
chlorination are compared, analysed and will be discussed in Chapter 4.
3.9.1
pH
pH value was used to express the intensity of the acid or alkaline condition of
a solution (Metcalf and Eddy, 2004). Figure 3.9 shows the pH meter used in this
study and pH measurement is a way to express the concentration of the hydrogen
ion. 30 mL of the wastewater sample was poured into the 50 mL conical flask and
measured with a pH meter. The pH electrode was rinsed with distilled water after
every usage and calibrated using standard solutions daily.
Figure 3.9: Meter pH model Orion 420 A
75
3.9.2
Biochemical Oxygen Demand (BOD)
BOD test provides a measure of the oxygen consumed in the biological
oxidation of organic (carbonaceous) material in a sample at 20ºC over a prescribed
period of time, usually 5 days. To date, BOD test is still the most popular parameter
for assessing the organic material strength of influents and effluents.
BOD
concentration was calculated according to Standard Method APHA 5210 (refer to
Appendix A) and by using BODTrak apparatus as shown in Figure 3.10.
Figure 3.10: BODTrak Apparatus
3.9.3 Chemical Oxygen Demand (COD)
COD test gives the electron donating capacity of practically all the organic
compounds in the sample, biodegradable or non-biodegradable and soluble or
particulate. A strong oxidizing agent, a hot mixture of dichromate and concentrated
sulphuric acid making chromic acid, oxidizes the organic compounds to CO2, the
same end product as biological activity (Knapp and Bromley-Challoner, 2003).
76
Some organic compound is not oxidized by the COD test but microorganisms
do utilize it. This organic material includes aromatic hydrocarbon (e.g. benzene and
phenols) and pyridines (water soluble benzene rings with an N in place of one of the
carbon atoms). They are mainly encountered in petrochemical wastewaters (Knapp
and Bromley-Challoner, 2003). COD value was determined using HACH DRB200
COD reactor (Figure 3.11) and HACH DR5000 Spectrophotometer which the
methods are included in Appendix B.
Figure 3.11: COD Reactor HACH DRB200
3.9.4
Ammoniacal Nitrogen (AN)
Nitrogen pollution in hydrosphere is attracting increasing attention for
eutrophication of lakes and rivers all over the world (Metcalf and Eddy, 2004).
Ammonium is the inorganic ion form of nitrogen pollution contained in municipal
sewage, industrial wastewater and agricultural wastes or decomposed from organic
nitrogen compounds in those wastewater and wastes.
Higher concentration of
ammonium will cause a sharp decrease of dissolved oxygen and obvious toxicity on
aquatic organisms (Wen et al., 2006). Concentration of ammonical nitrogen was
measured using HACH DR5000 Spectrophotometer (Figure 3.12) and the methods
are included in Appendix C.
77
3.9.5 Phosphorus
Concentration of phosphorus in sample was measured using HACH DR5000
Spectrophotometer (Figure 3.12) and the methods are included in Appendix D.
3.9.6 Tannin
Concentration of tannin in sample was measured using HACH DR5000
Spectrophotometer (Figure 3.12) and the methods are included in Appendix E.
Figure 3.12: HACH DR5000 Spectrophotometer
3.9.7 Escherichia Coli
Escherichia coli (E. coli) were determined by referring Standard Method
APHA 9260 as included in Appendix F. The instruments that will be used in
78
measuring E. coli were Quanty-Tray/2000 (Figure 3.13), Quanty-Tray Sealer (Figure
3.14 (a)) and Thermolyne Incubator (Figure 3.14 (b)).
Figure 3.13: Analysis of E. coli using Quanty-Tray/2000
(a)
(b)
Figure 3.14: (a) Quanti-Tray Sealer Model 2x and (b) Thermolyne Type 142300
Incubator
79
3.10
Disinfection of Effluent
In order to compare the bacterial removal, the chlorination process was
carried out in series of 800 mL beakers and stirred with magnetic stirrer machine
(Chemix model CL 6) to facilitate the reaction between the wastewater and chlorine
(Figure 3.15). This study will compare the state of Terminalia catappa leaf with
chlorination process to kill bacteria. Chlorination requires high cost compared to
Terminalia catappa leaf which is cheaper since it is easily to obtain and the process
of the producing the tannin extract is simpler. Moreover, there are quite a number of
Terminalia catappa trees in Malaysia.
Chlorine dosage ranging from 1.0 mg/L to 10.0 mg/L was used in the
analysis. Contact time between 10 to 30 minutes for each dosing were analysed to
obtain optimize timing for the reaction to occur. During the time interval, samples
were taken for E. coli test. Thus, the effectiveness of chlorination to kill the bacteria
can be measured directly from the E. coli measurement. The residual chlorine was
determined using Iodometric Method (Water and Wastewater Analysis, 2005) to
quantify the exact total residue in chlorination process.
Figure 3.15: Chlorination process using magnetic stirrer (Chemix model CL 6)
80
3.10.1 Iodometric Method for Determination of Residual Chlorine
Chlorination may produce adverse effects. In order to fulfill the primary
purpose of chlorination and to optimize any adverse effects, it is essential that proper
testing procedures be used with a foreknowledge of the limitations of the analytical
determination. In this study, Iodometric Method was used as a method to identified
and evaluate the residual chlorine in disinfection process by chlorination.
In this method, 25 g Na2S2O3.5H20 was dissolved in 1 L freshly boiled
distilled water and standardize against potassium bo-iodite and stored for at least 2
weeks. This initial storage is necessary to allow oxidation of any bisulfate ion
present. Make sure that use boiled distilled water was used and add a few mL
chloroform (CHCl3) to minimize bacterial decomposition. Then, standardize 0.1N
Na2S2O3 using iodate method.
For iodate method, 3.249 g KH(IO3)2 was dried at 103ºC for 1 hour, and
dissolved in distilled water and dilute to 1000 mL to yield a 0.1N solution. 80 mL of
distilled water then was added with 1 mL sulfuric acid. Then 10 mL 0.1N KH(IO3)2
and 1 g of KI was added in the water. Titration was conducted immediately with
0.1N Na2S2O3 titrant until the yellow colour of the liberated iodine was almost is
discharged. 1 mL starch indicator solution was added and titration was continued
until the blue colour disappears. Distilled water was used for titration as a blank
titration and the total residue was determined using the following equation;
mg Cl as Cl2 / L = ( A ± B ) x N x 35 450
mL sample
where:
A = mL titration for sample
B = mL titration for blank (positive and negative)
N = normality of Na2S2O3
(1)
81
3.11
Landscape Design
An open-air landscape garden of 1.5 m x 1.0 m x 0.3 m was designed and
constructed in front of Environment Engineering Laboratory, Faculty of Civil
Engineering, UTM as shown in Figure 3.16 and Figure 3.17. Five different plants
(Figure 3.18 and 3.19) was planted and covered the landscape, which are: Coleus
artopurpureus (Ca), Cyperus dubius Rottb (Cd), Nyctanthes arbo-tristis L (Na),
Panicaum maximum cv. Colonio (Pm) and Vinca rosea (Vr). Grass type
Elephantopus Scaber (Es) was also planted in the middle region of the garden. The
landscape garden was then watered everyday with 5 liter volume of treated effluent.
Another landscape garden was also constructed with the same types of plants and
served as control and it will be watered using untreated effluent.
Effluent Storage
Tank
Canopy
Soil
0.3m
1.5 m
SIDE VIEW
Figure 3.16: Landscape Garden Side View
82
Na
Cd
Va
Es
Es
Ca
Pm
Figure 3.17: Landscape Garden Plan View
Two storage tanks were located above the landscape garden where each of
them contains treated and untreated sewage effluent. The storage tanks were placed
on top of a hill to make sure the water flows easily to the plants. The plants were
watered every morning and evening by using spray conduit from the tank.
a)
b)
Figure 3.18: Landscape Garden with (a) Coleus artopurpureus (Ca) and (b)
Nyctanthes arbo-tristis L (Na)
83
a)
b)
Figure 3.19: Landscape Garden with (a) Five Different types of plants and (b) Vinca
rosea (Vr)
3.12
Plants Growth Determination
The plants growth rate was determined using two measurement techniques
adopted from Malaysia Agricultural Research and Development Institute (MARDI).
These techniques require the measurement of plants height and its leaves width
conducted weekly. The increment in both height and width indicate positive growth
rate. Figure 3.20 indicates the measurement of leaves width and Figure 3.21 indicates
the measurement of plants height.
84
Width
Determination
Figure 3.20: Width Growth Determination
Ruler
Higher Tree
Point
Ruler
Higher Tree
Point
Figure 3.21: Height Growth Determination
3.13
Preparation of Agar Nutrient (EMB Agar)
Bacteria are microorganisms that grow everywhere. We can collect and grow
them in specially prepared petri dishes. Blood agar or tryptic soy agar with 5%
sheep's blood is an excellent medium for supplying bacteria with nutrients and an
85
environment in which we can see them grow. Eosin methylene blue (EMB) agar was
prepared in this study for bacteria counting purpose. The agar was prepared by
dissolving 35 g of EMB media (Merck) into distilled water (Figure 3.22) and
autoclaved at 121°C for 15 minutes.
The media was then poured on disposable petri dishes and waited until the
agar was solidified in the laminar flow. The indigestible agar is a gelatin-like
substance with a semi solid surface on which the bacteria can grow while they
consume the added nutrients (like sheep's blood). Below are the steps that were
carried out;
1)
Prepared petri dishes were refrigerated and stored upside down (i.e
media in upper dish, cover on bottom). This keeps condensation
which forms in the lid from dropping onto and disrupting the bacteria
growing surface.
2)
When ready to use, the dishes were cooled to room temperature
before taking samples (about one hour).
3)
Without tearing the agar surface, inoculate the dish by gently pressing
using cotton bud or rod onto agar surface.
4)
Replace cover on dish, tape closed, and label each dish to know the
source of the bacteria. Store upside down.
5)
Let it grow in undisturbed warm location, ideally in an environment
around 100°F (37°C) - not in sunlight or on a heating register.
6)
Growth can be seen within a couple of days. The dishes will start to
smell which means bacterial are growing.
7)
Make observations and keep records of what is growing on each dish.
Make sure before conducting the work, ethanol is sprayed on the table
to make sure no bacteria is in the surrounding environment.
86
8)
Before disposing of dishes in the trash the bacteria should be
destroyed. Pour a small amount of household bleach over the colonies
while holding dish over sink.
a)
b)
Figure 3.22: Figure of (a) preparation of agar nutrient and (b) agar was poured in
petri dishes
3.13.1 Plate Colony Counting
The most frequently used method of measuring bacterial populations is the
plate count. An important advantage of this method is that it measures the number of
viable cells. One disadvantage may be that it takes some time, usually 24 hours or
more, for visible colonies to form (Gerard et al., 2004).
87
Plate counts assume that each live bacterium grows and divides to produce a
single colony. This is not always true because bacteria frequently grow linked in
chains or as clumps. In this study, the colonies that are form on the agar was counted
and the results recorded (Figure 3.23). If too many colonies are present, it indicates
that a lot of bacteria on the shrubs and serial dilution method is needed to ensure the
colony counts will be within range.
a)
b)
Figure 3.23: Figure indicates that the agar nutrient in petri dish (a) before and (b)
after counting the colonies (without bacteria)
3.13.2 Serial Dilutions for Plate Counting
When a plate counts is performed, it is important that only limited number of
colonies develop in the plate. When too many colonies are present, some cells are
overcrowded and do not develop; these conditions cause inaccuracies in the count
(Gerard et al., 2004). The Food and Drug Administration convention is to count
only plates with 25-250 colonies, but many microbiologists prefer plates with 30-300
colonies. To ensure that some colony counts will be within this range, the original
inoculum is diluted several times in a process called serial dilution (Figure 3.24).
88
For example, that wastewater has 10,000 bacteria per milliliter. If 1 mL of
this sample were plated out, there would theoretically be 10,000 colonies formed in
the petri dishes of medium. Obviously, this would not produce a countable plate. If
1 mL of this sample were transferred to a tube containing 9 mL of sterile water, each
milliliter of fluid in this tube would now contain 1,000 bacteria. If 1 mL of this
sample were inoculated into a Petri plate, there would still be too many potential
colonies to count on a plate.
Therefore, another serial dilution could be made. One milliliter containing
1,000 bacteria would be transferred to a second tube of 9 mL of water. Each
milliliter of the tube would now contain only 100 bacteria, and if 1 mL of the
contents of this tube were plated out, potentially 100 colonies would be formed and it
is easily countable. In this study, four dilution factors have been prepared to count
the colonies on the plants leaves.
Figure 3.24: Plate Counts and Serial Dilutions (Gerard et al., 2004)
89
3.14
Microbiology Sampling Procedures
Shrubs leaves were taken from the landscape garden three times a week to
measure the E. coli and during the sampling, the weather condition was observed.
Shrubs leaves of even size were picked later using sterile forceps and collected inside
centrifuge tubes. The forceps must be placed in oven for 24 hours to ensure no
bacteria around the forceps. Then, swab the leaves with cotton bud and swab on the
agar that have been prepared. The cotton bud (Figure 3.25 (a)), was heated for
several seconds on the fire (Bunsen burner) as shown in Figure 3.25 (b), to make
sure no effects of any bacteria from the environment.
The shining metallic-green colony on the EMB agar showed the presence of
E. coli in the wastewater. Then, count the colonies on the agar and the number of
colonies on plate must be multiplied with reciprocal of dilution of sample to get the
real number of bacteria in milliliter (mL).
a)
b)
Figure 3.25: Figure shows that (a) cotton buds that were used to swab the leaves and
(b) Bunsen burner that will used to heat up the cotton bud before use
90
3.15
Determination of NPK Ratio
A proper understanding of nitrogen-phosphorus-potassium (NPK) ratio will
ensure that crops are properly fed and nourished. These essential nutrients promote
vigorous growth, increased root development and improved disease and stress
tolerance. According to all plants surveyed, these three macronutrients are the most
popular nutritional elements required by plants, as they are all needed in the largest
volumes for healthy plant growth. Our top three are nitrogen (N), phosphorus (P) and
potassium (K). Like their three big brothers the primary macronutrients, secondary
macronutrients are also essential for healthy plant growth, but are needed in smaller
quantities for success. These three champions are calcium (Ca), magnesium (Mg)
and sulphur (S).
In this study, samples were taken from the soil before and after watering with
wastewater. The soil was digested using aqua-regia solution. Before the digestion,
the soil was heat in the oven for 24 hours. Then, the aqua-regia solution was
prepared by adding 5 mL of sulfuric acid and 15 mL of nitric acid into the beaker
(1HCL : 3HNO3). The solution was added with the soil from the different plots and
original soil and then was heated on the hot plate for 2 hours. The solution was then
filtered and analysed for total nitrogen, total phosphorus and potassium using HACH
DR5000 Spectrophotometer. Soil pH determination was also analysed for the initial
soil sample and soil taken from both landscape plots.
CHAPTER 4
RESULT AND ANALYSIS
4.1
Introduction
This chapter will discuss on the results that were obtained from the
experiment that had been done throughout the project. Data obtained were tabulated
in tables and shown in graphical approach for ease of analysis.
4.2
Optimum Dosage of Terminalia catappa (TC) Leaves
Process of determining the optimum dosage of Terminalia catappa leaves
was conducted using Jar Test method. In preliminary studies, jar test were conducted
to select suitable Terminalia catappa leaf condition or form that will produce
optimum tannin concentration. Researchers before only used TC leaf in its original
form and powder form for their study. However, this study would manipulate TC
leaves into blended with water. Thus, test was carried out to prove that TC leaves in
blended with water are better to be used than its origin condition including cut or
92
sliced condition. Analysis were conducted on the leaves in its original form, cut in 2
cm squares and blended with water.
4.2.1 Analysis of Tannin Concentration
Figure 4.1 shows the concentration of tannin obtained after 7 days of soaking
in distilled water for different forms of leaves. The test assessed the conditions of
the leaves; they were weighed in the range of 30 g to 70 g and added to 1000 mL of
distilled water each in a series of beaker. The test was run for 7 days and the
concentration of tannin and pH values were measured daily. From the figure, it
shows that tannin concentration increased with time for all the leaves condition. The
concentration of tannin for 70 g of leaves in blended form is 1932 mg/L followed by
1750 mg/L in cut form and 1696 mg/L in leave form.
Figure 4.1: Concentration of tannin from Terminalia catappa leaves in different
forms i.e. original leaf, cut and blended in distilled water
93
This proved that, Terminalia catappa leaves in blended form is better to be
used in treating wastewater because it would mix homogenously in water samples
and react with the samples quickly and effectively. The weight of 70 g Terminalia
catappa leaves gives optimum tannin concentration for all leaves form and this is in
accordance with previous study carried out on the leaves (Muhamad Hafiz Razi,
2009). According to Chansue, (2008) the total tannin level will increase with
increase in duration of extraction.
4.2.2 Analysis of pH measurement
From Figure 4.2, it can be seen that TC leaves shows a decrease in pH after 7
days of treatment. Leaves in blended form (70 g) shows the reduction in pH from
7.73 to 5.55 after soaking for 1 to 7 days. However, the cut leaves form was only
reduces from pH 6.98 to 5.85 after seven days. At 70 g, leaves in its original form
displays a slight decrease in pH i.e. a drops from 7.7 to 6.94 on the 4th day and
slowly reduced to 6.12 and decrease again to 5.85 on the 7th day. This happens
because the blended leaves are already in liquid form. Therefore tannin will mix
homogeneously with the distilled water and reduce the pH. This proved that TC
leaves in blended form are better to be used in treating wastewater because it would
mix easily in water sample where it would react with the sample quickly and
effectively.
94
Figure 4.2: pH measurement of TC leaves from 30 g to 70 g of immersion in
distilled water after 7 days
Figure 4.3: pH measurement for distilled water treated added with 70 g of TC leaves
in original, cut and blended form
95
4.2.3
Soxhlet Extraction
For comparison purposes medicinal extraction of tannin using Soxhlet
extraction was carried out using methanol as solvent. From the experiment high
concentration of total tannin was obtained for TC leaves at 50 and 70 g as shown in
Table 4.1. The results obtained shows high concentration of tannin extract compared
to conventional method of soaking the leaves in distilled water.
Table 4.1: Concentration of tannin and pH from Terminalia catappa leaves extracted
with Soxhlet apparatus
Weight of TC
Concentration
leaves (g)
(mg/L)
50
2.1 x 104
7.18
70
2.3 x 104
7.02
pH
Soxhlet extraction for 70 g of TC leaves yield 23 000 mg/L of tannin
compared to only 1932 mg/L from soaking method. Even though soxlet extraction
yields more tannin concentration it is deem unsuitable for high volume of wastewater
compared to conventional method due to solvent usage, time and electricity
consumption. Futhermore, the extraction procedure have to be monitored closely to
avoid accidental surge of solvent. The presences of methanol and other solvent
could also retard the growth of plants and increase the concentration of chemicals in
soil.
96
4.3
Wastewater Treatment Analysis
During the study, 25 liters of wastewater effluent were collected from the
oxidation pond once a week and analysed for pH, E. coli, BOD, COD, tannin,
ammoniacal nitrogen and phosphorus. Terminalia catappa leaves (1750 g) was then
blended with the effluent and then left to soak for five days in a tank. Samples were
then collected and analysed to ascertain its water quality based on Standard Method
for Water and Wastewater Analysis (2005).
4.3.1
pH
As expected a decrease in pH was observed for depicted domestic wastewater
after treatment with TC. Figure 4.4 shows a reduction in pH measurement after 5
days of treatment. pH level decreased from an average of pH 9.0 to pH around 6.0 to
7.0. The highest reduction was observed on 22nd May 2009 where pH dropped from
8.74 to 5.07.
Therefore, it is proven that Terminalia catappa leaves are capable of reducing
pH values in domestic. As we know, tannin are rich in phenolic compounds and are
capable of binding hydroxide ion (OH-) which may eventually effect the pH value in
wastewater treatment (Walton et al., 1999). According to Chris Yew (2004), too
much of Terminalia catappa leaves may result in too low water pH and the dried TC
leaves actually release organic acids like humic and tannins which lowers the pH of
the water. So whether you choose to use Terminalia catappa leaves to treat domestic
wastewater, it is more of a personal preference and a matter of convenience.
97
28-May
Figure 4.4: pH of domestic wastewater before and after treatment with TC leaves for
5 days
4.3.2
Biochemical Oxygen Demand (BOD)
Figure 4.5 below shows BOD result for the wastewater effluent before and
after treatment with tannin at various intervals of sample collection from 2nd May to
2th Jun 2009. From the figure, it shows that the values of BOD on 2nd May 2009 was
the highest value recorded and it shows a decreased from 39 mg/L to 32 mg/L after 5
days treatment. On 22nd May 2009, the lowest value for BOD was observed for the
wastewater at 9.9 mg/L and it was decreased to 7.6 mg/L after 5 days of treatment.
The removal efficiencies of BOD concentrations in treatment are 18%, 21%, 8%,
47%, 23%, 21% and 36% respectively from the 1st time sampling to the last event of
sampling.
98
28-May
28-May
Figure 4.5: Concentration of BOD in wastewater samples before and after treatment
with TC leaves for 5 days
From the graph, it can been seen that BOD concentration was reduced after
treatment with TC leaves after being immerse in wastewater for 5 days. This shows
that tannin is capable of reducing the numbers of microorganism and is capable of
degrading organic matter under aerobic conditions. In spite of this, sample that were
treated with Terminalia catappa leaves show the reduction in BOD concentration. It
indicates that the organic matter in sample with TC leaves were reduced after 5 days
and the blended form of TC leaves was able to treat BOD from domestic wastewater.
There is also a fluctuation in BOD concentration observed from the various sampling
intervals, this is however uncontrollable due to the nature of the study
99
4.3.3 Chemical Oxygen Demand (COD)
Chemical Oxygen Demand (COD) is one of the important parameters to be
analysed for domestic wastewater treatment. It is a measure of the total quality of
oxygen required to oxidize all organic material into carbon dioxide and water. Figure
4.6 shows the COD result obtained for all the wastewater samples collected from 2nd
May to 2nd Jun 2009.
From the various sampling intervals it shows that COD in the wastewater was
reduced after treatment with tannin except for sampling on 2nd Jun 2009. On 6th May
2009, COD concentration was147 mg/L and was reduced to 121mg/L after 5 days of
immersion with tannin. On 2nd Jun, there was a slight increase in COD concentration
from 256 to 297 mg/L. This increase was however uncountable for
28-May
Figure 4.6: Concentration of COD in wastewater samples collected on 2nd May to
2nd Jun 2009 before and after treatment with TC leaves
.
100
4.3.4
Ammoniacal Nitrogen
According to Figure 4.7, all domestic wastewater samples that were treated
with TC leaves displays removal for ammoniacal nitrogen after 5 days of treatment.
On 6th May 2009, 27% removal of ammoniacal nitrogen was observed. However on
other sampling intervals, small reduction of ammoniacal nitrogen concentration was
observed. An excellent removal efficiency of ammonia nitrogen was observed on
second set of treatment using Terminalia catappa leaves. On 6th May 2009, 27% of
removal efficiency was showed for ammoniacal nitrogen.
28-May
Figure 4.7: Concentration of ammoniacal nitrogen in wastewater samples collected
on 2nd May to 2nd Jun 2009before and after treatment with TC leaves
101
4.3.5 Phosphorus
Phosphorus levels were significantly decreased in all treatments using
Terminalia catappa leaves. The highest phosphorus concentration was recorded on
2nd Jun 2009 where the concentration on phosphorus was initially 44.2 mg/L and was
reduce to 30.2 mg/L after 5 days treatment (32%). From the graph (Figure 4.8), it
can be seen that the efficiency removal was achieved on 17th May 2009 which 33.6%
of removal was recorded.
28-May
Figure 4.8: Concentration of phosphorus in wastewater samples before and after
treatment with TC leaves
Phosphorus concentration in domestic wastewater is usually high due to fecal
matter and domestic products that contain phosphorus.
Alkaline conditions of
domestic wastewater may also induce high level of phosphorus being release in
wastewater. Joen et al. (2001) reported that in high pH, acetate (substrate) uptake
requires more energy compared to low pH environment. High energy requirement
leads to the high phosphate release due to hydrolysis of polyphosphate. Figure 4.8
102
shows the graph obtained for phosphorus concentration analysed for the wastewater
treated with TC leaves. From the data obtained, the concentration of phosphorus was
reduced for all the sampling intervals except on 6th May 2009. The reduction was
recorded highest on 2nd Jun 2009 i.e. from 44.2 mg/L to 30.2 mg/L.
4.3.6
Escherichia coli
Escherichia coli (E. coli) are indicator organisms that are used to indicate the
possible presence of pathogenic organisms associated with human and animal waste.
Pathogenic organisms are adapted to live in the host organism and, once out in the
environment, their numbers are reduced as a result of natural die-off, predation,
sedimentation, and an inability to adapt to higher or lower temperatures, ultraviolet
exposure, and unfavorable water chemistry.
Figure 4.9 shows E. coli numbers in domestic wastewater before and after
treatment with TC leaves. From the graph on 28th May 2009, 98% removal of E. coli
was observed. On 2nd Jun 2009, E. coli numbers were reduced from 2419 to 347
MPN. From the graph it could be observed that if the numbers of E. coli was more
than 1000 MPN, tannin was unable to entirely eliminate E. coli. The high contents
of tannins in Terminalia catappa leaves reveal that they may serve as a source of
natural antioxidants (Charng, 2002)
Tannins possess antibacterial properties (Chung et al., 1998). Tannic acid can
inhibit the growth of intestinal bacteria by binding with metal ions especially strong
binding with iron and then forming a chelate. The chelate, like a siderophore, is
toxic to the membranes of microorganisms. When tannins form chelating complex
with iron in the medium, this action makes no iron available for microorganisms to
grow under aerobic condition therefore, bacterial growth was inhibited (Scalbert,
103
1991). E. coli growth was inhibited by tannins only when tannins were exposed to
oxygen (Alexandra et al., 2003).
28-May
Figure 4.9: E. coli numbers in domestic wastewater effluent before and after treated
with TC leaves after 5 days
4.4
Analysis of Coliform on Leaves and Grass
Two landscape plants were constructed with five different plants were
planted to cover the area.
The landscape plants being applied with treated
wastewater with Terminalia catappa and the other one was watered using untreated
wastewater (control). Coliform analysis was also carried out on leaves and grass
from the landscape garden. The results for E. coli presence in five different plants
from different plots were determined. However, the same pattern of occurrence for
E. coli found on plants as in wastewater where bacteria presence on experimental
plants is less than the control plants. Nevertheless, both plants on the landscape
gardens show the presence of bacteria. This is probably because some coliform
104
bacteria occurs naturally on the plants and contributed to the positive results in E.
coli determination (Francis and Frederick, 2000).
4.4.1
Coleus artopurpureus (Ca)
Figure 4.10, shows the presence of E. coli on Coleus artopurpureus (Ca)
plant at the landscape garden. From the graph, it can be seen that the presence of
coliform was higher on 17th May 2009 for both treated and untreated landscape plots.
It might be due to the weather on that day where it was raining throughout the day.
As we know, bacteria prefer to grow without UV irradiation and at the same time, the
minerals and nutrients present in the soil might contribute to bacteria survival and
growth (Mohd Ismid et al., 2003). The landscape plant that was constructed was also
covered by canopy which can obstruct the plants direct exposure to UV irradiation
(sunlight).
Figure 4.10: E .coli on Coleus artopurpureus (Ca) plant during different
sampling time intervals
105
4.4.2
Cyperus dubius rottb (Cd)
Figure 4.11, displays the presence of E. coli on Cyperus dubius rottb (Cd)
plants for different landscape plots. From the graph, it indicates that high of coliform
numbers were present on 17th May 2009. However, the coliforms that were observed
after 23th May 2009 are much lower. This could be due to the surface area of the
plants are smaller compared to other plants such as Coleus artopurpureus (Ca) which
have large leaves area (Figure 4.12). From the result, it can be summarized that the
surface area of the leaves play important role in order to provide shade from direct
sunlight and offer suitable condition for bacteria survival and growth.
Figure 4.11: E. coli on Cyperus dubius rottb (Cd) plant during different
sampling time interval
106
a)
b)
Figure 4.12: (a) Cyperus dubius rottb (Cd) plant and (b) Coleus artopurpureus (Ca)
with different surface leaves area
4.4.3
Nyctanthes arbo-tristis L (Na)
Figure 4.13 shows the presence of E. coli on Nyctanthes arbo-tristis L (Na)
from the two different landscape plots. It is apparent from that plants with treated
wastewater contain less coliforms compared to untreated wastewater. It also indicates
the presence of coliforms on the leaves that could be due to the natural condition.
On 25th May 2009, it was observed that the lower number of coliforms
present on the Nyctanthes arbo-tristis L (Na) plants. It was only 12 CFU/mL for
treated landscape plots compared 23 CFU/mL from untreated landscape plots. This
might be due to the concentration of tannin extracted from Terminalia catappa
leaves which acted as disinfecting agent in order to kill bacteria. Moreover, the
leaves also release humic acid. Tannic and humic acids may be useful for inhibiting
many types of bacteria including cyano-bacteria (Yew, 2004). As observed, the
107
highest number of coliforms presences was on 17th May 2009 which could be due to
the weather condition on that day as mentioned earlier.
Figure 4.13: E. coli on Nyctanthes arbo-tristis L (Na) plant during different
sampling time interval
4.4.4
Panicaum maximum cv. colonio (Pm)
The results in Figure 4.14 indicate that most of the coliforms from untreated
landscape plot contain more coliforms compared to the treated wastewater.
Generally, coliforms result is more reliable in determining the contamination brought
by watering using sewage effluent since it excluded the influences of natural bacteria
(Hilal et al., 2003).
108
It was observed that same result was obtained on 17th May 2009 for the
number of coliforms present to the plants. From the result, it displays that the
untreated landscape plants contains 1900 CFU/mL of coliforms on the leaves
compared to treated landscape plants which only contains 800 CFU/mL of coliforms
on the Panicaum maximum cv. colonio (Pm) leaves. The results also might be due to
the sunlight effect through to the plants. In the landscape plants, the Panicaum
maximum cv. colonio (Pm) was planted at the edge of the landscape plots.
Figure 4.14: E. coli on Panicaum maximum cv. colonio (Pm) plant during different
sampling time intervals
From this side, the plants were directed to the sunlight and as we know,
higher UV light irradiation resulted in lower survival rate, and vice versa. This might
due to the fact that the UV emissions penetrate the cell wall of microorganisms,
prevents cell replication and causes the cell to die (Metcalf and Eddy, 2004).
109
4.4.5
Vinca rosea (Vr)
Figure 4.15, shows that the number of coliforms presence on Vinca rosea
(Vr) plant after being watered with treated and untreated wastewater. Due to the
result, the graph was plotted and displays the same result with the other plants where
the numbers of coliforms are much higher on 17th May 2009. However, Vinca rosea
(Vr) plants show the lowest number of coliforms for treated plots compared to other
plants in the same plots.
From the observation Vinca rosea (Vr) plant for treated landscape plots have
more radiant and shining flowers which are violent in colour compared to the control
plants. This might be due to the tannin concentrations that are rich in Terminalia
catappa leaves. As we know, TC is good in release tannins and humic acid. Tannins
are polyphenolic compounds and acts as anti-bacterial agents. It also might interact
and produce enzymes that create radiant and shinning flowers (Mueller-Harvey,
2001).
Figure 4.15: E. coli on Vinca rosea (Vr) plant during different sampling time
intervals
110
4.4.6
Elephantopus scaber (Es)
Elephantopus scaber (Es) is a type of grass that was planted in the middle
region of landscape plot to cover the area. From the Figure 4.16, it displays that the
number of coliforms present on the plant after watering with treated wastewater
using Terminalia catappa. On 17th may 2009, the numbers of coliforms are much
higher compared to other days for both landscape plots. It was recorded that the
number of coliforms on 17th May 2009 for treated landscape plots was 1700
CFU/mL compared to untreated landscape plots at only 900 CFU/mL of coliforms.
Figure 4.16: E. coli on Elephantopus scaber (Es) plant during different sampling
time intervals
On 25th May 2009, it was observed that the lower number of coliforms
present on Elephantopus scaber (Es) was as same as for Nyctanthes arbo-tristis L
(Na) plants. It was only 3 CFU/mL for treated landscape plots compared 1 CFU/mL
from untreated landscape plots. This might be due to the shady conditions of the
grass, receiving less UV irradiation compared to the plants. In this condition, the
111
survival rate of fecal colifom is higher as the amount of UV irradiation was
insufficient to kill the bacteria as mentioned before (Mohd Ismid et al., 2003).
4.5
Analysis of Coliform on Soil from Landscape Plots
The results in Figure 4.17 indicate that most of the coliforms remained inside
the soil sample. From the graph, it shows that soil from treated wastewater plot
contains less coliform bacteria compared to untreated effluent plot. From the figure,
it shows that the number of coliforms presence was up to 48 000 CFU/mL for
untreated soil compared 32 000 CFU/mL for treated soil. The number shows that
there are quite a large numbers of coliforms compared to the one on the plants. The
lowest numbers of coliforms was recorded on 2nd Jun 2009 where the number was
670 CFU/mL for control plots compared to treated landscape plot at only 520
CFU/mL.
The difference in coliform numbers was due to the effectiveness of TC leaves
that were used in this study. The presence of coliform in the soil was much higher
than on the plants. This could be due to the fact that soil are located under the shade
of the plants and was not exposed to direct sunlight, thus receiving less UV
irradiation compared to the plants samples. It is possible that the UV irradiation on
the soil samples was not strong enough to kill the bacteria (Mohd Ismid et al., 2003).
Furthermore, nutrient and minerals present in soil may also contribute to the bacteria
growth and survival.
112
Figure 4.17: E. coli in soil samples analysed on 11th May to 2nd June 2009
4.6
Wastewater Influence on Plant Growth
Table 4.2 shows the average growth rate of plants for experimental and
control landscape garden. It can be seen that shrubs applied with wastewater grew
faster compared to untreated wastewater with1-2 fold increment in growth of plants
height and width depending on types of plants with an exception for Elephantopus
scaber (Es). One major factor that contributes to the faster growth of the plants is the
presence of organic fertilizing agent such as nitrogen, phosphorus and potassium in
the wastewater (Mohd Ismid et al., 2003). As we know, plants requires chemical
elements (nitrogen, phosphorus and potassium) in order to provide better growth as
they are all needed in the largest volumes for healthy plant growth. They also
required macronutrients such as calcium, magnesium and sulphur to support their
growth rate.
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Table 4.2: Average growth rate of plants from experiment and control landscape
garden
Shrubs
Coleus
artopurpureus
Control – growth
rate (%)
Leave
Height
Width
Experiment – growth
rate (%)
Leave
Height
Width
34.7
105.9
53.2
125.0
24.9
-
41.3
-
-
410.3
-
292.0
48.4
98.5
73.3
128.5
Panicaum maximum
cv. colonio
72.9
76.9
90.0
78.6
Vinca rosea
61.6
52.9
95.7
170.6
Cyperus dubius rottb
Elephantopus scaber
Nyctanthes arbotristis L
In this study, it was observed that the flowers for treated wastewater plot was
more radiant and shining compared to the control plants. This could be due to
reaction of polyphenolic compound in tannin forming enzymes to brighten the plants
colour (Charles, 2005). Elephantopus scaber (Es), which is a type of grass that have
growth rate in control plots rather than experimental plots where the grass grow up
faster and creep on the garden. This could be due to the plants preference for pH ~ 7
(Walton et al., 1999). Elephantopus scaber (Es), in the control samples prefer
neutral pH to growth where they cannot grow up in lightly acidic condition.
Referring to the pictures in Figure 4.18, tannin and humic acid may be
consequence why the flower becomes more shinning and radiant. According to
World Intellectual Property Organization (2009), tannin may be further used as antiparasitic properties which were found to greatly reduce the palatability of plants
susceptible to parasite attacks. The result was due to the synthesis of polyphenols by
plants where secondary metabolites, such as phenols and by-products often have the
114
function of inhibiting the palatability of the parts of the plant wherein they are
produced and stored in terminal buds of plants.
a)
b)
Figure 4.18: (a) Flowers watered with wastewater and TC is more radiant and
shining compared to (b) control plants
According to Yew (2004), humic acid contains sulphur, nitrogen and
phosphorus in varying amounts. It also contains metals such as Ca, Mg, Cu and Zn
which can be chelated in some undefined way. These nutrients are important and the
plants required all the nutrients in order to support their growth rate. Humic acid can
be broken down into two groups based on the polarity and size of the individual
compounds. The smaller, more polar fraction is generally termed fulvic acid and the
larger; more non-polar fraction is generally termed humic acid. Humic acids are the
end product of microbial degradation of plant and animal debris and are one of the
most important constituents to fertilize soils. In addition, tannic and humic acids
may also be useful for inhibiting many types of bacteria that might affect the plants
growth rate.
115
pH values may also effects the growth and radiance of the flowers. The soil
is the foundation of any garden where healthy soil will lead to a healthy garden and
plants. The sign of earthworms is also a good indicator for soil health. Lime can be
added to soil with low pH (acid) to raise the pH to a suitable level. Most plants
prefer pH around 6.0 to 7.5 (Charles, 2005). Table 4.3 shows the result of pH value
for each soil from different plots of landscape garden. The original soil indicates
acidic soil with 5.68 pH value. Soil samples collected form experimental and control
plot have pH reading of 6.96 and 7.04 respectively. This shows that wastewater was
capable of neutralizing acidic soils and could proved to be beneficial for landscape
gardens
Table 4.3: pH values for different soil samples collected from landscape garden
Sample
pH value
Original Soil
4.77
Soil with Treated Wastewater
6.96
Soil with Untreated Wastewater
7.04
Soil pH is one of the most important soil properties that affect the availability
of nutrients. As we know, macronutrients tend to be less available in soils with low
pH and micronutrients tend to be less available in soils with high pH (Charles, 2005).
In the suitable pH range, nutrients are more readily available to plants, and microbial
populations in the soil increase. Microbes may convert nitrogen and sulphur to
forms that plants can use. Lime also enhances the physical properties of the soil that
promote water and air movement
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4.7
Growth Rate of Plants
Growth rate of plants was measured based on guidelines for measurement
from Malaysia Agricultural Research and Development Institute (MARDI, 2007).
The measurement was conducted using measurement tape and meter.
4.7.1
Growth Rate of Vinca rosea (Vr)
Figure 4.19 and 4.20, it shows that the increments in plant height and leaves
width for Vinca rosea (Vr) plant in experimental plot and control plot.
The
increment in both height and width indicates positive growth rate. From the figures,
it can be seen that increase height and width was observed for both the experimental
and control plots.
Treatment with TC shows higher increment in height and width compared to
control plants. 95.7% in height was measured for this plant in the experimental plot
compared to 61.6% in the control plant. The growth percentage for leaves width for
Vinca rosea (Vr) is 170.6% for experimental plots compared to 52.9% for control
plots.
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29-May
Figure 4.19: Growth rate of Vinca rosea (Vr) plant in terms of height for
experimental plot and control plot garden
29-May
Figure 4.20: Growth rate of Vinca rosea (Vr) plant in growth of width (leaves) for
experimental plot and control plot garden
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4.7.2
Growth Rate of Coleus artopurpureus (Ca)
Figure 4.21 and 4.22, it displays that the result of growth rate for Coleus
artopurpureus (Ca) plant in both height and width of leaves. From the figure, it
shows that the plant with untreated wastewater indicates slightly higher growth rate
compared to the plants with treated wastewater. From the graph, it shows that the
incremented of height of plant indicate from 1st May 2009 to 29th May 2009. The
measurement of both height and width was conducted once a week.
29-May
Figure 4.21: Growth rate of Coleus artopurpureus (Ca) plant in terms of height for
experimental plot and control plot garden
The control samples show the positive increments compared to experiments
sample could be due the reason that started from the beginning of the project, the
plants were bought from the “Pak Ali Nursery” in Taman Pulai Flora, Skudai Johor.
Plants that are bought from the nursery are larger in size and have been grown for
several days at the nursery. For Coleus artopurpureus (Ca) plant, the height of the
control plants was higher than to the experiment plants when it was planted in the
landscape plant. As a result, the height and width of leaves for the control plant
show better growth rate compared to experiment plant. However, the growth rate for
119
the experiment samples in width of leaves show an increment after 1 month due to
the effect of tannin concentration in Terminalia catappa leaves.
29-May
Figure 4.22: Growth rate of Coleus artopurpureus (Ca) plant in terms of width
(leaves) for experimental plot and control plot garden
4.7.3
Growth Rate of Nyctanthes arbo-tristis L (Na)
Nyctanthes arbo-tristis L (Na) plant was planted in both landscape plot and
the growth rate was measured. From the graph in Figure 4.23 and 4.24, it shows the
same result such as for Vinca rosea (Vr) where the experiment samples indicates
positive growth rate compared to control sample. As we know, using domestic
wastewater in order to watered the plants may provide more nutrient i.e. nitrogen and
phosphorus. The control sample also shows the incremented of growth rate after one
month but was much lower compared to experimental.
120
29-May
Figure 4.23: Growth rate of Nyctanthes arbo-tristis L (Na) plant in terms of height
for experimental plot and control plot garden
29-May
Figure 4.24: Growth rate of Nyctanthes arbo-tristis L (Na) plant in terms of width
(leaves) for experimental plot and control plot garden
121
4.7.4
Growth Rate of Panicaum maximum cv. colonio (Pm)
Figure 4.25 and 4.26 shows the result of growth rate in both height and width
of leaves for Panicaum maximum cv. colonio (Pm) in different landscape plots. Both
height and width indicates positive growth rate after one month being applied with
treated and untreated wastewater. However, on 22th May 2009, the leaves width it
for control sample was slightly higher compared to experiment samples. The leaves
width for experiment samples shows that increment due to the addition of wastewater
and tannin provides nutrients to the plants. pH value for the soil sample may also
affect the result and provide better growth rate to the plants.
29-May
Figure 4.25: Growth rate of Panicaum maximum cv. colonio (Pm) plant in terms of
height for experimental plot and control plot garden
122
29-May
Figure 4.26: Growth rate of Panicaum maximum cv. colonio (Pm) plant in terms of
width (leaves) for experimental plot and control plot garden
4.7.5
Growth Rate of Cyperus dubius rottb (Cd)
In this study, plant types Cyperus dubius rottb (Cd) was selected to be
planted in the landscape garden. In measuring the increments of growth rate, only
height of the plants was measured using the techniques adopted from MARDI
(2007). This might could be due to the plant are difficult to measure the leaves width
where the surface area of the leaves were to small (Figure 4.27). Based on the graph
in Figure 4.28, only small increment occurred in height of Cyperus dubius rottb (Cd)
plant. This might be due to the fact that the plants did not prefer to grow in
conditions where the pH value was less than 7. The type of soil also influences the
growth rate of the plant whether it is sandy or peat area.
123
29-May
Figure 4.27: Cyperus dubius rottb (Cd) plant with small leaves surface area
29-May
Figure 4.28: Growth rate of Cyperus dubius rottb (Cd) plant in terms of height for
experimental plot and control plot garden
124
4.7.6
Growth Rate of Elephantopus scaber (Es)
As mentioned earlier, Elephantopus scaber (Es) in the growth rate was lower
growth compared to control samples. This measurement was carried out only on the
width of the plant because the plant creeps on the soil of the landscape garden. This
type of grass was planted in the middle of the region of landscape plants in order to
cover the area. From the graph, it shows that on 29th May 2009, the control samples
recorded 59.7 cm of width compared to experiment control which was only 44.3 cm.
29-May
Figure 4.29: Growth rate of Elephantopus scaber (Es) plant in terms of width for
experimental plot and control plot garden
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4.8
Nitrogen-Phosphorus-Potassium (NPK) Ratio
Complex fertilizers contain nitrogen (N), phosphorus (P) and potassium (K),
(NPK) in a 4-3-3 ratio (Shaheen et al., 1998). Plants need different nutrients at
different stages of their growth. In the root stage they need what are called root
fertilizers or potassic fertilizers. In the growing stage they need “leaves and foliage
fertilizers” (nitrogenous fertilizers). In the flowering and fruit stage they need “fruit
and flowering fertilizers” (phosphates fertilizers). In the experiment, the result of
NPK ratio may indicates the positive growth rate for the plants. Table 4.4 below
shows the result of the experiment. From the result, soil treated with wastewater
shows more nutrients where the NPK ratio was 641:31:1 followed by soil with
untreated wastewater at 622:25:1 and original soil at 582:31:1. This shows that
application of sewage effluent for landscape plants provide more nutrients for plants
or shrubs to grow.
Table 4.4: NPK ratio of different soil in landscape garden
Sample
Total
Total
Potassium
Nitrogen
Phosphorus
(mg/L)
NPK Ratio
(mg/L)
(mg/L)
Original Soil
9 900
530
17
582:31:1
Soil with Treated
13 800
650
18
641:31:1
11 200
450
18
622:25:1
Wastewater
Soil with Untreated
Wastewater
As mentioned earlier in subtopic 4.6 above, presence humic acid may
increase the concentration of nitrogen, phosphorus and sulphur. Therefore, using
Terminalia catappa leaves in order to water the plants provide more nutrients to the
plants which contain more tannin and humic acid. Therefore, the leaves have more
advantageous since it exhibit antibacterial properties and provide more nutrients for
126
plants. Furthermore, with tannin concentration may make the flowers more radiant
and shining could be due to reaction of polyphenolic compound in tannin forming
enzymes as mentioned above.
4.9
Disinfection
In order to achieve bacterial removal in effluent, sodium hypochlorite was
used as an oxidizing agent to disinfect the wastewater (Abdullah, 1994). From
experiments carried out, different chlorine dosage and contact time was found to
bring different disinfection effect as reported by White (1992). Table 4.5 shows
bacteria removal increases with contact time.
Table 4.5: Effect of chlorine dosage and contact time towards E. coli survival rate
Chlorine
E. coli
dosage
MPN / 100ml
(mg/L)
10 mins
20 mins
30 mins
1.0
2419.6
1011.2
347
3.0
1120.1
745
263
5.0
573
0
0
7.0
0
0
0
10.0
0
0
0
Longer the contact time leads to more efficient the disinfection process. The
chlorine dosage at 5 mg/L for contact time of 20 minutes was considered to be
optimum in this study where all E. coli was removed from the wastewater. From the
result obtained, it shows that disinfection process also may be applied in order to kill
127
bacteria where E. coli was reduced from 2419.6 MPN to 347 MPN after 30 minutes
in 1 mg/L of chlorine dosage.
Figure 4.30: Bacteria removal after disinfection process with different chlorine
dosage and contact times
In order to save the treatment time and cost, the chlorine dosage used in
previous study was 2 mg/L for contact time of 30 minutes contact times (Mohd Ismid
et al., 2003). However, the application of chlorine is known to have residual effect.
Table 4.6 and Figure 4.31 shows the total residue from chlorination measured using
Iodometric Method.
From the result at 5.0 mg/L chlorine dosage 6.31 mg/L chlorine residue was
recorded whereas, at 10 mg/L chlorine dosage, the residual chlorine was 128.5 mg/L.
This shows that even though chlorine is effective in eliminating bacteria it has
residue which might be harmful to human, plants and insects.
128
Table 4.6: Total residue of chlorination using Iodometric Method
Dosage (mg/L)
Total residue ( mg/L)
1.0
0.94
3.0
2.99
5.0
6.31
7.0
11.08
10.0
128.5
Figure 4.31: Chlorine residue after disinfection process with different chlorine
dosage
CHAPTER 5
CONCLUSION AND RECOMMENDATION
5.1
Conclusion
Water is one of the most important resources for all living things on earth. In
the 21st century, serious environmental problems such as pollution triggered much
concern about the relationship between human and environment. There are various
causes to water pollution either from point source or non-point source pollution.
Clean water is getting harder to obtain either for daily usage or in irrigation sector.
Reuse effluent from domestic wastewater have been widely used worldwide for
irrigation sector.
Furthermore, there are countries that reuse the effluent from
domestic wastewater especially countries experiencing scarcity in their natural water
resource. United States and Australia are two representative countries that reuse the
effluent for daily activities such as irrigation and toilet flushing. Therefore, the
treatment for water resources is important to ensure the public health.
The first objective of this study is to identify the antibacterial properties of
Terminalia catappa leaf. From the results and analysis, it can be concluded that the
objectives of the study was achieved. Terminalia catappa leaves were effective in
treating wastewater. It produces tannic acid which has antibacterial properties that
130
can treat or eliminate E. coli. From the result obtained all samples that were treated
using TC leaves displays removal of E. coli numbers at 98% after 5 days of
treatment. Based on the result, it can be concluded that the number of E. coli will
decrease when water is more acidic and TC leaves is capable of destroying the
bacteria. In other words, the water extracts of Terminalia catappa leaves have
potential to be use as an alternative for antibacterial agents and chemical substances.
Based on water quality parameters analysis, Terminalia catappa leaves
showed the positive results such as reduced in pH of sample from an average of pH
9.0 to 6.0 and 7.0 respectively. The reduction occurred due to tannic acid mixing
with the wastewater. The blended form of Terminalia catappa leaves also can
reduce BOD, AN, COD and phosphorus in domestic wastewater.
The results also showed that further removal of bacteria can be achieved if
the effluent is disinfected with chlorination. Obviously, the disinfection will also
make the effluent safer for landscape workers and the public. In order to compare
bacterial removal in effluent with chlorine disinfection, results shows that 5 mg/L of
chlorine was capable of eliminating E. coli after 20 minutes. However, chlorine has
residual effect and is harmful to plants, human and insects. The treatment with TC
will make domestic effluent safer for landscape workers and eventually the public.
In reusing effluent wastewater for specific uses it is imperative that reuse be
made safe to the public. At present, different countries have different standards that
are applied to wastewater reuse for the purpose of landscape garden irrigation.
Among them is Australia with fecal coliform <2000 MPN/100 ml and US EPA with
fecal coliform < 14 MPN/100 mL. Therefore, it is important that any attempt to
explore the potentiality of wastewater reuse in Malaysia must be preceded by the
establishment of a safe standard and regulation for its use.
131
The result of this study has shown that treated sewage effluent applied to
landscape plants produce better growth compared to untreated wastewater. The
results obtained showed positive growth increment for wastewater with TC leaves in
terms of height and width for all plants except for Elephantopus scaber (Es) which
thrive more when watered with wastewater.
This could be due to the plants
preference for pH ~ 7.
We can conclude that tannin can be further used as anti-parasitic properties
which were found to greatly reduce the palatability of plants susceptible to parasite
attacks.
The result was due to the synthesis of polyphenols by plants where
secondary metabolites, such as phenols and by-products often have the function of
inhibiting the palatability of the parts of the plant wherein they are produced and
stored in terminal buds of plants. It was also observed that the flowers watered with
wastewater and TC is more radiant and shining compared to the control plants. pH
values may also effects the growth and radiance of the flowers. The soil is the
foundation of any garden where healthy soil will lead to a healthy garden and plants.
The sign of earthworms is also a good indicator for soil health. The results of this
study have also shown that the number of coliform decreased by using wastewater
treated with Terminalia catappa leaves and decreased due to upon exposure to
sunlight in the landscape garden
5.2
Recommendation
Reusing wastewater has been practiced for years. The applications treatment
of Terminalia catappa for domestic wastewater treatment is able to provide water
quality suitable for reuse. Suggested domestic wastewater-reuse activities such as
irrigation, toilet flushing, and vehicle washing do not require high water quality
standard. Most saliently, the return of cleaned water back into the ecosystem close
132
to where it is used helps to increase aquifer recharge and river flow, allowing for
more natural cleansing through the river system.
Terminalia catappa can be used to treat wastewater.
There are certain
improvements that can be done to improve the study for better results.
The
improvements and modifications can be carried out from times to times to keep
abreast of the latest technologies available. Some of the reccomendations to improve
this study are as below:-
(i)
Analysis on other parameter such as Total Suspended Solid (TSS) in
order to check the blended form of Terminalia catappa leaves may
turbid the wastewater or may
provide clogging problem to the
treatment system. Beside, other parameter i.e. zinc and ferum also
must be analysed to check the efficiency of Terminalia catappa
leaves in treating heavy metals.
(ii)
Use more advanced extraction method to increase tannin extraction of
the leaves instead of using Soxhlet extraction which have high solvent
usage, time and electricity consumption. By using the extraction
method, problem of suspended solids may be overcome
(iii)
Use a sprinkler to sprayed the effluent to the landscape garden to
assess the extent of bacterial spreading during before and after
watering
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APPENDIX A
DETERMINATION OF BODTrak
USING HACH DR5000 SPECTROPHOTOMETER
145
146
147
148
149
150
APPENDIX B
DETERMINATION OF CHEMICAL
OXYGEN DEMAND (COD) USING HACH DR5000
SPECTROPHOTOMETER
152
153
154
155
APPENDIX C
DETERMINATION OF AMMONIACAL
NITROGEN USING HACH DR5000
SPECTROPHOTOMETER
157
158
APPENDIX D
DETERMINATION OF PHOSPHORUS
USING HACH DR5000 SPECTROPHOTOMETER
160
161
APPENDIX E
DETERMINATION OF TANNIN USING
HACH DR5000 SPECTROPHOTOMETER
163
164
APPENDIX F
E. COLI DETERMINATION
166
167