INTRODUCTION

Water is an unusual compound with unique physical properties. As a result, it is the

compound of life. Yet, it is the most abundant compound in the biosphere of Earth. However,
due to its capability to dissolve many substances, water is rarely found in pure form and contains
other substances physically or chemically, and also microorganisms. All organisms require
water. Water helps dissolves nutrients in food and helps transfer it to the whole body. Materials
that harmful to water quality are known as or pollutant contaminants. Contaminants in water
usually cause little harm to water as they are not lethal and are at low concentration. Pollutant
substances that dissolve in water however can cause harm to water even at very low
concentration.

WQI is a dimensionless number that combines multiple water-quality factors into a

single number by normalizing values to subjective rating curves. Factors to be included in
WQI model could vary depending upon the designated water uses and local preferences. Some
of these factors include DO, pH, BOD, COD, total coliform bacteria, temperature, and
nutrients (nitrogen and phosphorus). These parameters occur in different ranges and expressed
in different units. The WQI takes the complex scientific information of these variables and
synthesizes into a single number. Formula to calculate WQI is :

WQI = (0.22 x SIDO) + (0.19 x SIBOD) + (0.16 x SICOD) + (0.15 x SIAN) + (0.16 x SISS) +
(0.12 x SIpH)

Escherichia coli, or E-Coli, is a common bacterium that can be found in diverse

environments all over the planet, including the gastrointestinal tracts of animals and humans.
Many of these strains of E-Coli are essential mechanisms in the digestive tract, while others are
pathogens that can cause complications in urinary and intestinal tracts. (Payne & Sparks) In
research, E-Coli is commonly used as a model organism, meaning they are widely studied by
scientists for a variety of purposes due to their experimental advantages. E-Coli is comparatively
simple, and there are many advantages to studying these prokaryotic cells in the fields of
biochemistry and molecular biology. E-Coli has this simplicity and is relatively easy to
propagate in a lab environment. In other points, E. coli, is one of the most common species of

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fecal coli form bacteria. It is a normal component of the large intestines in humans and other
warm-blooded animals, and it’s found in human sewage in high numbers. E. coli is used as an
indicator organism for fecal contamination because it is easily cultured. If sewage is present in
water, pathogenic or disease-causing organisms may also be present.

Theory for e-coli clearly state that, there are 4 type of phase in growth of bacteria until
the bacteria death. The microbial culture in batch culture system (shake flask system) goes
through a lag phase, exponential growth phase, decelerating growth phase, stationary phase and
sometimes the death phase depends on the end product desired. The substrate concentration in
the culture medium and growth parameters, such as glucose concentration changes
correspondingly throughout the growth phases.

BASIC CONCEPTS
Water quality is the condition of the water body or water resource in relation to its
designated uses. It can be defined in qualitative or quantitative terms. Parameters in defining
water quality can be grouped into three board categories: physical, chemical, and
biological. Physical factors include temperature, sediment and bed material, suspended
sediments, turbidity, color, and odor. Chemical factors consist of the major and minor
elements, and other chemical parameters such as pH, Dissolved Oxygen (DO), Biological
Oxygen Demand (BOD), and Chemical Oxygen Demand (COD). The major elements include
agro-nutrients such as Nitrogen and Phosphorus; and minor elements include elements such as
arsenic (As), lead (Pb), and mercury (Hg). Biological Constituents include Fecal Coli-form
and E-coli. Conventionally water quality is expressed in terms of the measured value of one or
more of these parameters in relation to their accepted or implied limits. They are expressed in
different units, and their magnitudes can vary significantly from one location to another and
over time. For example, the temperature is expressed in degrees Celsius or degrees Fahrenheit,
and coliforms in numbers, and most chemicals and nutrients in milligrams per liter (mg/L) or
in parts per million (ppm).

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OBJECTIVES

These experiments is carried out :

● To determine water quality index for the prepared water sample.
● To determine the dissolved oxygen (DO) level in the lake water sample.
● To ascertain whether the lake water sample comply with Malaysian Water Standards
based on the Water Quality Index.
● To identify the factors affecting the concentration of dissolved oxygen in the lake water
sample.
● To detect and confirm of total coliform and E-Coli in water.

APPARATUS

E-COLI APPARATUS

Reagent: Colitag Presence Water Sample

100ml Measuring Cylinder E-Coli Bottle

3
 Clear Coliplate E-Coli
Coli Apparatus

Sealer Sealer Machine

Lamp U/V Portable Insert and Most Probable

Number (MPN) Table

4
WATER QUALITY INDEX APPARATUS

Water Sample 25 ml Measuring Cylinder

Mineral Stabilizer Polyvinyl Alcohol Dispersing

Nassler Reagent Pipette Syringe

5
Test Tube with Cap Spectrophotometer

Dilution Water Phosphate Buffer Solution

Ferric Chloride Solution Calcium Chloride Solution

6
Magnesium Sulphate Solution Dissolved Oxygen Meter

pH Meter Incubator

7
PROCEDURES

E-COLI EXPERIMENT

1) 110mL of water samples was poured into the E-Coli bottle by measured it using the
100mL measuring cylinder.
2) The reagent which is Colitag Presence was put into the E-Coli bottle that already
contained the 110mL of water samples.
3) The bottle was closed tightly and the bottle was shake to let the reagent and water
samples mixed together.
4) Then, the mixtures of water sampleswere put into the clear and empty Coliplate.
5) The Coliplate was then seal by using the sealer and sealer machine so that the mixture of
water samples does not spill.
6) After the Coliplate was sealed completely, then it was put into the E-Coli Apparatus.
7) The Coliplate was left for 24 hours of minimum duration time before take it out.
8) Then, by using the lamp U/V Portable, the blue color of small and large weir that
appeared was calculated.
9) Then, by using the Insert and Most Probable Number (MPN) Table, the number of
microorganisms was obtained.

WATER QUALITY INDEX EXPERIMENT

1) 3 drops of mineral stabilizer were put into two measuring cylinder of 25mL, then 3 drops
of Polyvinyl Alcohol Dispersing were put into the same measuring cylinder.
2) 1mL of Nassler Reagent was added into the same measuring cylinder by using pipette
syringe.
3) The distilled water was added into the first measuring cylinder until the volume reached
25mL. The water sample was added into the second measuring cylinder until the volume
reached 25mL.
4) The measuring cylinder was shake until all the solution dissolved completely and was let
it to rest for 1 minutes.
5) Then the samples in both measuring cylinder was poured into the two test tube with cap.
6) Then the samples in the test tube was put into spectrophotometer to find the concentration
in each samples and the data of the samples was recorded.

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Water Sample without dilution

1) Another water sample was put into empty beaker to find it concentration and pH by using
the Dissolved Oxygen Meter and pH meter.
2) The DO meter was switched on. The dissolved oxygen was tested by using DO meter.
3) The probe of the DO meter was put into the bottle and the ‘mode’ button was pressed.
4) The data recorded when the beep sound was heard.
5) For pH data, pH meter was turned on.
6) The electrode was rinse with distilled water under an empty beaker then it was put into
the water samples.
7) ‘READ’ button was pressed to get the pH of the water samples.
8) The reading was record which was the true data for pH of the water samples.

Water Sample with dilution

1) 50 ml of another water samples being measured by using measuring cylinder.
2) The measured 50ml of water sample was poured into two BOD bottle.
3) The BOD bottle was filled up with dilution water that consists of mixturePhosphate
Buffer Solution,Ferric Chloride Solution, Calcium Chloride Solution andMagnesium
Sulphate Solutionuntil it full and put the stopper. Make sure that there was no bubble in
the measuring cylinder.
4) The BOD bottle was labelled as and .
5) The DO meter was switched on. The dissolved oxygen was tested by using DO meter.
6) The probe of the DO meter was put into bottle and the ‘mode’ button was pressed.
7) The data recorded when the beep sound was heard.
8) The BOD bottle that labelled as was kept in the incubator for 5 days.
9) After 5 days, steps 6 until 7 was repeated for bottle.

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RESULT AND ANALYSIS

E-COLI EXPERIMENT

Locations Of Water Samples Taken Tasik Seksyen 7

Volume Of Water Sample Used (ml) 110
E-Coli Type Escherichia coli β-glucuronidase positive
Reaction Time (hour) 24
Incubator Temperature (°°c) 35
Number Of Blue Colonies On Small Weir 48
Number Of Blue Colonies On Large Weir 49
Positive
Number Of Microorganism (MPN) >2419.6

WATER QUALITY INDEX EXPERIMENT

Biochemical Oxygen Demand (BOD)

Time Start: 10.30PM

SAMPLE VALUE
Dilution Water (Litre) 1
Sample Bottle Volume, Vb (mL) 300
Sample Volume, Vs (mL) 50
Decimal Volumetric Fraction of Sample Used (P) 0.17
Initial Dissolved Oxygen, DO0 (mg/L) 7.43
Final Dissolved Oxygen, DO5 (mg/L) 6.11
Biochemical Oxygen Demand at 5 days, BOD5 (mg/L) 7.76

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Sample Calculation for BOD:

( )
P =
( )

= 0.1667 ≈ 0.17

! "#$% &' ! "#$%

BOD5 =
( ) ( ') ( * +

,- &,-
=
.

/.0 &1.22
=
.2/

= 7.76 mg/L

Volatile Suspended Solids

The weight of the dish (mg) 1148.8
The weight of solids + dish before ignition (mg) 1250.0
The weight of solids + dish after ignition (mg) 1172.2
Difference of before and after ignition (mg) 77.8
Volume of sample (ml) 20

Sample calculation for Volatile Suspended Solids:

(7& )×2
Volatile Solids (mg/L) =
!

Where:
A = weight (mg) of solids + dish before ignition
B = weight (mg) of solids + dish after ignition

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 (29 . &22/9.9)×2
Volatile Solids =
9
= 3890 mg/L

( &:)×2
Fixed Solids (mg/L) =
!

Where:
B = weight (mg) of solids + dish after ignition
C = weight (mg) of dish

(22/9.9&220;.;)×2
Fixed Solids =
9
= 1170 mg/L

(<&=)×2
Suspended Solids (mg/L) =
!

Where:
A= weight (mg) of solids + dish before ignition
C = weight (mg) of dish

(29 . &220;.;)×2
Suspended Solids =
9
= 5060 mg/L

Location Of Water Samples : Tasik Seksyen 7

PARAMETER
Sampling Ammonia TSS DO BOD pH COD WQI Class
concentration
(mg/L) (mg/L) (mg/L) (mg/L)
(mg/L)
Water 2.08 5060 3.04 7.76 6.28 50 36.03 IV
Sample

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CLASS OF WATER SAMPLES ACCORDING TO EACH PARAMETER

Parameter Value Class

DO 3.04 mg/L III
BOD 7.76 mg/L IV
COD 50 mg/L III
NH3-N 2.08 mg/L IV
TSS 5060 mg/L V
pH 6.28 II

CALCULATION FOR WQI

WQI=(0.22 × @A ) + (0.19 × @A ) + (0.16 × @AD ) + (0.15 × @AEF) + (0.16 ×

@A@@) + (0.12 × @AGH)

SUBINDEX COEFFICIENT;

SUBINDEX FOR DO (IN % SATURATION) :

For x ≤ 8, SIDO = 0

SUBINDEX FOR BOD :

For x > 5, SIBOD = 108 × exp(−0.055N) − 0.1N

= 108 × exp(−0.055 ∗ 7.76) − 0.1(7.76)

= 69.704

SUBINDEX FOR COD :

For x > 20, SICOD = 103 × exp(−0.0157N) − 0.04N

= 103 × exp(−0.0157 ∗ 50) − 0.04(50)

= 44.980

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SUBINDEX FOR NH3-N :

For 0.3 < x < 4, SIAN = 94 × exp(−0.573N) − 5 ∗ |N − 2|

= 94 × exp(−0.573 ∗ 2.08) − 5 ∗ |2.08 − 2|

= 28.144

SUBINDEX FOR SS :

For x ≥ 1000, SISS = 0

SUBINDEX FOR pH :

For 5.5 ≤ x < 7, SIpH = −242 + 95.5N − 6.67N 9

= −242 + 95.5(6.28) − 6.67(6.28)9

= 94.686

Hence,

WQI = (0.22 × 0) + (0.19 × 69.704) + (0.16 × 44.980) + (0.15 × 28.144) + (0.16 × 0) +

(0.12 × 94.686)

= 36.03

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DISCUSSION

This experiment was carried out to monitor and do assessment on the water sources that
is important in order to make sure that the water sources used is clean and not polluted. Members
of two bacteria groups, coliforms and fecal streptococci, are used as indicators of possible
sewage contamination because they are commonly found in human and animal feces. Although
they are generally not harmful themselves, they indicate the possible presence of pathogenic
(disease-causing) bacteria, viruses, and protozoans that also live in human and animal digestive
systems. Therefore, their presence in streams suggests that pathogenic microorganisms might
also be present and that swimming and eating shellfish might be a health risk. In addition to the
possible health risk associated with the presence of elevated levels of fecal bacteria, they can also
cause cloudy water, unpleasant odors, and an increased oxygen demand.

This experiment was conducted by using the multiple-tube fermentation method involves
adding specified quantities of the prepared water sample, which is taken from TasikSeksyen 7 to
tubes containing a nutrient broth. Yellowish solution was formed after the mixing. Then, the
solution was poured into the colilert and sealed to be put into the incubator under 35°C for the
period of 24 hours to get the positive result. Positive results that occur in less than 24 hours are
valid, but the results cannot be recorded as negative until the 24-hour incubation period is
complete. After the incubation, the colilert were scanned by using the long wave UV lamps to
determine the presence of Escherichia coli β-glucuronidase positive.

Thus, from the scanning using the long wave UV lamps, the number of small weir that
becomes blue under the scanning process is 48, while the of large weir that turns into the blue
color under the scanning is 49. Blue color indicates that the water is slightly polluted and there is
microorganism in the water samples. From the IDEXX Quanti-Tray/2000 MPN Table per 100ml,
the number of microorganism that is determined in the water is slightly greater than 2419.6.

Next experiment is done to determine the water quality index for the prepared water
samples. Besides, Water Quality Index (WQI) is used to classify the class and status of river.
There is six parameter in order to determine the water quality index which is dissolved oxygen
(DO), biological oxygen demand (BOD), chemical oxygen demand (COD), Ammonical

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Nitrogen (NH3-N), total suspended solids (TSS) and pH value. The value of WQI depends on the
value of the sub index for each parameter.

For the first parameter, the dissolved oxygen that we got is 3.04mg/L. Dissolved oxygen
refers to the level of free, non-compound oxygen present in water or other liquids. It is an
important parameter in assessing water quality because of its influence on the organisms living
within a body of water. A dissolved oxygen level that is too high or too low can harm aquatic life
and affect water quality. Then, we got 7.76mg/L for biological oxygen demand (BOD).
Biological oxygen demand is a measure of the quantity of oxygen used by microorganisms such
as aerobic bacteria in the oxidation of organic matter. BOD directly affects the amount of
dissolved oxygen in rivers and streams. The greater the BOD, the more rapidly oxygen is
depleted in the stream. This means less oxygen is available to higher forms of aquatic life. The
consequences of high BOD are the same as those for low dissolved oxygen: aquatic organisms
become stressed, suffocate, and die.

The concentration of chemical oxygen demand(COD) for prepared water samples is

50mg/L. Chemical oxygen demand (COD) is the amount of oxygen necessary to oxidize all of
the organic carbon completely to CO2 and H2O.Chemical Oxygen Demand is an important water
quality parameter because, similar to BOD, it provides an index to assess the effect discharged
wastewater will have on the receiving environment. Higher COD levels mean a greater amount
of oxidizable organic material in the sample, which will reduce dissolved oxygen (DO) levels. A
reduction in DO can lead to anaerobic conditions, which is deleterious to higher aquatic life
forms. Next parameter is the concentration of ammonical nitrogen in the water samples. Thus,
the ammonical nitrogen concentration was determined to be 2.08 mg/L by mixing the water
samples with the 3 drops of ammonia solution and 1ml of Nessler Reagent. Ammonia is one of
several forms of nitrogen that exist in aquatic environments. Unlike other forms of nitrogen,
which can cause nutrient over-enrichment of a water body at elevated concentrations and indirect
effects on aquatic life, ammonia causes direct toxic effects on aquatic life.

Furthermore, the total suspended solid (TSS) of the water samples is 5060 mg/L. Total
suspended solids (TSS) is the dry-weight of suspended particles, that are not dissolved, in a
sample of water that can be trapped by a filter that is analyzed using a filtration apparatus.
Higher total suspended solid could lead to the higher level of water turbidity which determined

16
that the water is polluted. Lastly, the pH of the water samples is found to be 6.28 which
determined that water is quite acidity. The term pH stands for “potential for hydrogen.” It’s a
measure of the concentration of hydrogen ions in a diluted solution. Acidic solutions contain
more hydrogen ions. And alkaline solutions contain fewer. Normal drinking water is at pH 7
which indicates as neutral and safe to be used.

Hence, from the calculation using the WQI formula which is,

WQI=(0.22 × @A ) + (0.19 × @A ) + (0.16 × @AD ) + (0.15 × @AEF) + (0.16 ×

@A@@) + (0.12 × @AGH)

We got 36.03 which classified as class IV according to the DOE Water Quality Index
Classification table. This class IV water is suitable to be used as irrigation. From the WQI
calculation, the water could not be determined as clean or polluted because the value is out of the
range that was given by the DOE Water Quality Index. Lastly, we could determined that Total
suspended solids parameter could lead to the value of Water Quality Index that we get as it has
bigger value that could give impact on the WQI value.

CONCLUSION

The water sample that was taken from Tasik Seksyen 7 are to be used for the testing of E-
Coli and determining of Water Quality Index of the water sample. Escherichia coli, also known
as E-coli, is a Gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus
Escherichia that is commonly found in the lower intestine of warm-blooded organism. Based on
this experiment it can be conclude that the colilert were scanned by using the long wave UV
lamps to determine the presence of Escherichia coli b-glucuronidase positive. Dissolved oxygen
analysis measured the amount of oxygen dissolved in an aqueous solution. Oxygen gets into the
water by diffusion from the surrounding air by rapid movement (aeration) and also as a waste
product of photosynthesis process. Dissolved oxygen analysis can be used to determine the
health or cleanliness of a lake or stream, the amount and type of biomass a freshwater system can
support and the amount of decomposition occurring in the lake or stream. Based on the dissolved
oxygen value that we get, we can determine the Water Quality Index of the sample water lake.
Water quality Index is parameter of water quality measurement used while INWQS or national

17
water quality standard is the classification from thus parameter result valued basis from certain
maximum permissible concentration or limits. Hence from the test, it could conclude that the
calculated water quality index is 36.03 which classified as class IV according to the DOE Water
Quality Index Classification table. Therefore, the objectives for both two experiments which are
E. coli and water quality index (WQI) experiment are accomplished and achieved.

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