Table of Contents

Abstract........................................................................................ 2
Objectives..................................................................................... 2
Introduction................................................................................... 3
Theory........................................................................................... 4
Procedure...................................................................................... 7
Results.......................................................................................... 8
Graphs...................................................................................... 10
Sample Calculations.................................................................10
Discussion................................................................................... 11
Sources of Error/ Limitations:....................................................13
Conclusion..................................................................................14
Appendix..................................................................................... 15
Questions.................................................................................... 16
Bibliography................................................................................ 18

Abstract
The following lab report provides a detailed analysis of three
experiments typically used in determining the quality of water while
comparing it to international water quality standards. The experiment
performed and reported on was the bacterial analysis of water from various
sources including: the laboratory tap, river water treated with household
bleach and an untreated river water sample. The second experiment
investigated in this report involves determining the likely sources of the
provided water samples. This is done by analyzing the physical and chemical
characteristics of the water samples. The third test known as the Jar Test
investigates the optimum coagulant dosage required to treat a particular
water on a laboratory scale. The results and analysis for this report is
presented through tabulated results, plotted graphs

and a detailed

discussion which relates obtained results to theoretical expectation and their

real world expectations.

Objectives

To assess the bacteriological quantity in water samples from the

following sources:
1. Laboratory tap
2. River water treated with household bleach
3. Untreated River water
To determine the optimum coagulant dosage under laboratory
conditions by use of the Jar Test to treat water from treatment plant.
2

To determine the sources of the three provided water samples using

chemical based tests

Introduction
Water has many unique properties that allow it to be used for a wide
variety of purposes. Some obvious uses would include drinking water,
cooking with water, washing, and even transportation as via ships and
barges. Water has excellent thermal properties. It can be heated; even to the
point of converting it to a gas we call steam. It can be cooled, providing the
air conditioning we enjoy in office buildings. Finally, unlike many of our fossil
fuels, it is plentiful), and it's relatively cheap. It is for these very reasons that
water must be treated to ensure that it doesnt harms its users.
An essential test carried out during the treatment and testing of water
is the bacteriological analysis test using membrane filter technique. This
involves using various sources of water to examine the amount of bacteria
present within them. This makes use indicator organisms to indicate the
presence of harmful micro-organiasm This technique is used mainly in water
and wastewater treatment to ensure that the disease carrying organisms are
identified and prevented from infiltrating the public water supply. This allows
a check for the water quality entering the public water supply which must
meet the World Health Organisation (WHO) standards.
Another test used to analyze water quality is the jar test. The jar test is
a common laboratory procedure used to determine the optimum coagulant
doses, in a laboratory setting which can be used to determine the coagulant
dosage of a large-scale treatment process of raw water from a similar source.
These impurities in raw water are typically suspended, which leads to
turbidity, odor and poor taste.. In order to remove these particles, they must
agglomerate and grow in size in order to settle out of solution. Therefore to
3

obtain agglomeration a chemical coagulant such as alum is used. This

causes the suspended particles to agglomerate and form flocs, which are
then heavy enough to settle and removed.
Source determination is a method used by water treatment companies
to determine the origins of a water leak or unidentified water body. Each and
every water source has varying physical and chemical characteristics. The
determination of each characteristic allows for a water treatment company
to determine the source. These physical and chemical characteristics include
pH level, turbidity, hardness, odour, the level of alkalinity, chloride and
chlorine concentrations. The values obtained from the experiments for these
physical and chemical characteristics are compared to the expected values
to determine the source.

Theory
BACTERIOLOGICAL ANALYSIS BY THE MEMBRANE-FILTER TECHNIQUE
The membrane--filter technique is now considered the preferred
method for the bacteriological analysis of water, unless the turbidity of the
sample is sufficient to clog the pores of the filter. It permits: the examination
of a larger volume of sample unlike other methods.
A given quantity of water is filtered through a porous disk of polyester
cellulose acetate. The size of the pores is 0.45 microns (u). Since coliform
bacteria are from 1 to 4 microns in length, they are retained on the filter
when the water passes through. Because the pores of the filter are so small,
the water will not pass through the filter by gravity. Therefore, a vacuum
source must be available to enable the water to penetrate the filter. After the
4

water sample has been filtered, the filter is incubated at 35C for 22-24 hours
in contact with M--Endo medium in a petri dish. This culture medium is
favorable to the growth of coliform bacteria, but it inhibits the growth of
other types of bacteria. the coliform bacteria will produce readily identifiable
colonies.

Colonies

may

be

counted

by

the

naked

eye.(

http://armymedical.tpub.com/MD0160/MD01600099.htm)

Jar Testing
A large percentage of the particles in surface and groundwater cannot
be removed by sedimentation because they are colloidal particles. These are
extremely small particles (particle sizes generally range from 1 nm to 1 m)
which have a negligible mass and large surface area per unit volume,
therefore, they tend to acquire a negative charge.
Colloidal and suspended particles are the primary cause of turbidity in
water. The turbidity of a body of water is defined as the insoluble (colloidal)
particles which impede the passage of light through water. It is measured in
Nephelometric Turbidity Units (NTU) and is used to quantify the degree to
which light traveling through a column of water is scattered by the
suspended organic and inorganic particles.
In water treatment, the process of coagulation, flocculation and
sedimentation is used to effectively remove colour and many organic
molecules and particles from the water. Coagulation consists of adding and
mixing a chemical reagent (coagulant/flocculant) to the raw water in order to
destabilize the suspended colloidal particles. One of the major coagulants
used in the treatment of turbidity in surface water is Aluminum sulphate
(Alum). The process of flocculation involves slow stirring to encourage the
clustering of the destabilized particles formed during coagulation resulting in
the formation of heavy, rapidly settling flocs. The destabilized particles are
then removed by sedimentation and filtration.

Jar testing is a method of simulating a full scale water treatment process,

providing system operators a reasonable idea of the way a treatment
chemical will behave and operate with a particular type of raw water.
5

Because it mimics full-scale operation, system operators can use jar testing
to help determine which treatment chemical will work best with their
systems raw water. The treatment chemical used in this lab experiment is
Alum. The Jar testing entails adjusting the amount of treatment alums added
to samples of raw water held in jars. The sample is then stirred so that the
formation, development and settlement of floc can be watched just as it
would in the full scale treatment plant. Floc forms when the treatment
chemical reacts with material in the raw water and clump together. The
operator then performs a series of tests to compare the effects of different
alum dosages at different pH values and turbidity to determine the right size
of

floc

for

particular

plant.

(http://www.nesc.wvu.edu/pdf/dw/publications/ontap/2009_tb/jar_testing_dwf
som73.pdf)
Source Determination
Water from varying sources will always possess the physical and
chemical characteristics typical of the environment in which originated.
Therefore there are several tests that can be performed in the laboratory
setting to identify these unique characteristics in order to indicate the
possible source of a water sample. The tests carried out in this experiment
include: pH tests, test for residual chlorine, hardness, alkalinity and chloride.
Ph Test:
The pH scale measures how acidic or basic a substance is. So
essentially the pH of a substance is a measure of hydrogen ion activity within
the substance It ranges from 0 to 14. A pH of 7 is neutral. A pH value above
7 indicates an alkaline substance and a ph values below 7 indicates an acidic
substance. Raw water is ideally treated to attain a pH value as close as
possible to 7 as a value other than 7 indicates the presence of contaminants
such as chemical and industrial wastes. The two methods used to measure
pH are:
6

1. Visual- using pH strips which turn a particular colour when placed in the
sample. This is then compared to a chart of colours which gives the
corresponding pH value.
2. Electrometric involves the use of a pH meter
Residual Chlorine
Chlorine and chlorine-based disinfectants are used worldwide to destroy
pathogens in potable water. One of the reasons for the widespread use of
chlorine disinfectants is that they provide a residual level of protection
against waterborne pathogens. A chlorine residual is a low level of chlorine
remaining in water after its initial application. It constitutes an important
safeguard against the risk of subsequent microbial contamination after
treatment which is a unique and significant benefit for public health.
(

http://www.waterandhealth.org/chlorine-residual-public-health-

safeguard/#sthash.9W6m036y.dpuf)
Hardness
Many industrial and domestic water users are concerned about the hardness
of their water. Hard water requires more soap and synthetic detergents for
home laundry and washing, and contributes to scaling in boilers and
industrial equipment. Hardness is caused by compounds of calcium and
magnesium, and by a variety of other metals. Water is an excellent solvent
and readily dissolves minerals it comes in contact with. As water moves
through soil and rock, it dissolves very small amounts of minerals and holds
them in solution. Calcium and magnesium dissolved in water are the two
most

common

minerals

that

make

water

"hard."

(https://www.fcwa.org/water/hardness.htm)
Alkalinity

Alkalinity is a measure of the capacity of water to neutralize acids. It can also

be referred to as the capacity to accept H+ ions. Alkali substances in water
include hydroxides or bases. Alkalinity is often related to hardness because
the main source of alkalinity is usually from carbonate rocks (limestone)
which are mostly CaCO3. If CaCO3 actually accounts for most of the
alkalinity, hardness in CaCO3 is equal to alkalinity.
Chloride:
The presence of chloride ions in natural waters can be attributed to
various sources such as discharges from effluents from chemical industrial
plants, oil well operations and sea water intrusion in coastal areas. These can
all cause the contamination of both surface and groundwater. Chlorides are
present in in potable water supplies as well as sewage, usually as a metallic
salt. A high chloride concentration in water is not known to have any toxic
effect on human beings. However, it can act corrosively on metal pipes and
be harmful to plant life.
When sodium is present in water, chloride
concentrations of greater than 250 mg/L cause a detectable salty taste.
Therefore, the standard of a maximum chloride concentration of 250 mg/L in
drinking water has been established more for reasons of taste than as a
safeguard against any physical risk.

Procedure.
Experiment 1 Jar Test
Equipment

was

replaced

during

the

experiment

due

an

equipment

malfunction. Following the replacement the experiment was carried out

following the procedural steps listed in the Laboratory Manual.
Experiment 2 Jar Test
The experiment was carried out following the procedural steps listed in the
Laboratory Manual.
Experiment 3 Source Determination
The experiment was carried out following the procedural steps listed in the
Laboratory Manual.

Results

Table 1: Bacterial Analysis

Table 2: Jar Test

9

Table 3: Source Determination

Graphs

Alum Dosage vs Turbidity

Sample Calculations
Experiment 1-Bacteriological Analysis(Sample C)
Coliformcolonies for 100 ml of sample=

of colonies counted
100
volume of sample filtered

10

Total Coliform colonies for 100ml of sample C=

5
100=5000000
0.0001

Fecal Coliform colonies for 100 ml of sample C=

3
100=3000000
0.0001

Experiment 3 Source Determination (Sample X)

Hardness of sample X =

Titre value1000
5.81000
mg
=
=232
of CaCO3
Vol of sample tested
25
L

Alkalinity of sample X=

Titre value0.0250000 150.0250000

mg
=
=150
of CaCO3
Volof sample tested
100
L

Titre value500
4.1500
mg

Chloride of sample X=
=
=82
of CL
Vol of sample tested
25
L

Discussion
In Bacterial Analysis experiment, three water samples were tested to
determine whether they were safe or unsafe for drinking. This was done
using the membrane-filtration method. These samples tested were:

Sample A- Tap water laboratory tap

Sample B Treated river water (household bleach)
Sample C Untreated river water

The membrane-filtration method in keeping with the theoretical

expectations, gave a direct count for each sample as reported in Table 1. In
the case of sample C (Untreated river water), the bacterially population was
expected to be high due to the nature of the sample. Therefore, in order to
obtain an accurate determination of the organisms present, the volume of
11

the sample was put through a serial dilution procedure in which successive
volumes of the sample are further diluted until the desired volumes of the
test sample are obtained.
The number of total coliform and fecal coliform colonies in the three
samples of water were used as indicators for presence of other potentially
harmful bacteria that typically strive under the same conditions. The
acceptable coliform standards (refer to Table 1 in Appendix) expressed in
colonies/100 ml of water sample varies depending on the use of the water.
For drinking purposes, however, the number of total coliforms must not be
greater than 1 total coliform. Although a 0 total coliform number doesnt
necessarily indicate that the water is safe to drink.
Sample A can be considered bacteriologically safe as there were no
detectable colonies of total coliform or fecal coliform. This was an expected
result as the sample was taken from the laboratory tap which is considered
to be the end result of the water. Water can only be considered an end result
after it has been treated to acceptable standards. Also the water was
expected to contain residual chlorine to prevent the formation and growth of
micro-organisms during transmission.
Sample B can be considered bacteriologically safe as there were no
detectable colonies of total coliform or fecal coliform. This proves that
treatment of water with bleach is an effective disinfecting process which
capable of removing bacteria from an otherwise contaminated source.
For Sample C high amounts of total and fecal coliform were detected.
There were 5,000,000 total coliforms and 3,000,000 fecal coliforms for the 1
ml sample and 630,000,000 total coliforms and 120,000,000 fecal coliforms
in the 10 ml sample. These high values indicated that there is likely a high
concentration of other harmful bacterial typically found alongside the
coliform.

12

In experiment 2, the jar test procedure was performed to simulate the

coagulation, flocculation and sedimentation processes that would be
undertaken in a water treatment plant for particular water conditions, which
can vary depending on the source. In this case, the source is raw river water.
Theoretically as the alum dosage increased, the turbidity of the
samples should decrease. However, the results of the jar test contradicted
this. As the alum dosage was increased the turbidity increased to a
maximum value and then began to decrease. From the graph, the optimum
coagulant dose, which defined as the dose at which there was the largest
change in turbidity was 63 mg/L.
The Source determination experiment was carried out to determine of
the source of three treated water samples. The three water samples were
taking from the following sources:

Treated Water from a salt water aquifer.

Treated water from a Desalination Plant
Treated water from the Caroni Treatment Plant

From the results it can determined that sample X was likely taken from
a salt water aquifer. This conclusion can be drawn based on the higher
chloride content of 82 mg/l present in sample X compared to sample Y and Z.
It also has the highest hardness value of 232 which is likely to present in
groundwater due to high calcium and magnesium sources in the ground.
The source of sample Y was determined to be water from a salt water
desalination

plant.

Product

water

from

desalination

plants

is

characteristically low in mineral content, hardness, alkalinity, and pH.

Therefore, desalinated water must be re-mineralized prior to final distribution
and use. The low hardness of 36, low alkalinity of 22 and low pH of 7.52
conforms to what is expected in desalinated water.

13

Lastly, the source of sample Z was deemed to be the treated water

from the Caroni Treatment plant. This is supported by the fact that its
chloride and hardness levels, at 44 mg/L and 100 mg/L respectively, are
within the range of that recommended for potable water. The strong
presence of residual chlorine indicates that the water sample has been
treated with chlorine, which is typically done in the end stages of the
treatment process. The pH level was found to be 7.15 which is typical for
drinking water which has an acceptable pH range between 6.5 to 8.5.
Unfortunately a high degree of confidence cannot be placed in the
results obtained for this experiment due to numerous mishandling of the
equipment and poor pipetting of the sample and the buffer.

Sources of Error/ Limitations:

Experiment 1 Bacteriological Analysis

The possible contamination of the membrane filter by external bacteria

not relevant to the experiment. This may have been facilitated by the
transfer forceps, especially if used incorrectly.
Possible contamination of samples due to the switching of the
malfunctioning equipment.
Incorrect use of the pipettes which could result in wrong volume
measurements of samples thereby giving inaccurate results. This is
especially relevant in the case of sample C where the serial dilution
technique was employed.

Experiment 2 Jar Test

The turbidity level in each jar was not the same before the coagulant
was added, therefore resulting in varying rates of change in turbidity.
Improper use of pipette to measure applying alum to the water
samples
Not allowing enough time for floc to settle
Smudging of samples when putting into the turbidimeter.

Experiment 3

14

Titration process visually based and therefore subjective to the person

performing the test.
Not enough sample tests were done to get an average result thereby
increasing the accuracy.

Conclusion
The bacteriological quality of the three water samples were analyzed
using the membrane-filtration method. It was found that the water from the
laboratory tap and the river water treated with bleach were bacteriologically
safe as no detectable coliform colonies were found. However, the river water
contained a vast number of coliform and therefore it can be considered
unsafe to consume.
The optimum Alum dosage for a water treatment plant treating river water
63 mg/L.
In the source determination experiment, the various tests were
effectively used to determine the most likely source for each sample:

Sample X was determined to be water from a salt water aquifer.

Sample Y was determined to be water from a salt water desalination

plant.

Sample Z was determined to be from the treated water of the Caroni

Treatment plant.

Appendix

15

Questions
16

1. In the titration, the phenolphthalein indicator changes from pink to

colourless at a pH 8.3 when acid is added to it. Water that has a pH
greater than 8.3 is said to have phenolphthalein alkalinity (P) which is
due mainly to the presence of carbonate or hydroxide ions. In this
experiment, the water samples had no phenolphthalein alkalinity (i.e. P
= 0) and therefore remained colourless after adding the indicator.
Therefore, from Table 2 (refer to Appendix), when P = 0, T = HCO3-.
All three water samples have Total Alkalinity in the form of
bicarbonates.
2. The rationale behind testing for coliform bacteria is that Pathogens like
protozoa, and viruses that make people sick can be rare and difficult to
detect even if they are present in the water. Coliforms are indicators and are
more common and easy to grow. Testing for them provides a margin of safety.
Pathogens may not be present if coliforms are, but it would be wise to look for
problems just in case.

In order to measure 0.001mL of a very contaminated sample, the serial

dilution process is used and proceeds as follows:
Put 1 ml of contaminated sample into dilution bottle 1

dilution)
Put 1 ml from bottle 1 into dilution bottle 2

0.0001ml dilution)
Put 10 ml form

bottle

through

the

(1/100

(0.01 * 1/100 =
membrane

filter

(0.0001*10)= 0.001 ml of sample

3. A water quality index provides a single number that expresses overall

water quality at a certain location and time based on several water
quality parameters. Some of the parameters used include biological
oxygen demand, total coliform bacteria and nutrients. The objective of
an index is to turn complex water quality data into information that is
understandable and useable by the public.
17

From the results in experiment 1, the fecal coliform concentration was

found to be 30000 colonies/100 ml in Sample C bottle 1 -1ml and
12000 in Sample C bottle 1-10ml. The Q-values according to the Water
Quality index is 7 and 20 respectively.
4. The measurement of Turbidity is an important test when trying to
determine the quality of water. It is an aggregate optical property of
the water and does not identify individual substances; it just says
something is there. Limiting turbidity prevents increases in water
temperatures because suspended particles absorb more heat. This, in
turn, reduces the concentration of dissolved oxygen (DO) because
warm water holds less dissolved oxygen than cold. Limiting turbidity
also reduces the amount of light penetrating the water, which reduces
photosynthesis and the production of dissolved oxygen.

Turbidity in

groundwater is mostly inorganic particles (silt, clay and natural chemical

compounds like calcium carbonate) and caused by natural geological factors.
Turbidity in groundwater can be removed by means of filtration. And the

process of coagulation.
5. Similarities:

In all three graphs there is a clearly defined region where the is a

large change in turbidy vs The coagulant dosage.

Differences:
In both of the graphs included in the appendix it can be seen that
the turbidity initially decreases as the alum dosage is increased
but in the graph obtained from the experiment the opposite
occurred. The turbidity initially increased with the increase in
coagulant dosage.

18

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Brown Robert M, McClelland Nina, Deinninger Rolf A and Tozer Ronald. Water and
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Water Stewardship Information Series. Hardness in Groundwater. Accessed
October
14,
2015.
http://www.env.gov.bc.ca/wsd/plan_protect_sustain/groundwater/library/ground_fact
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United Nations Environment Programme.Desalination by
Accessed:
October
15,
http://www.oas.org/usde/publications/Unit/oea59e/ch20.htm

reverse

osmosis.
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Hardness of Water by EDTA Titration. Accessed: October 16, 2015.

http://cd1.edb.hkedcity.net/cd/science/chemistry/s67chem/pdf/sOL_6_Water_hardne
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19

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