WOLLEGA UNIVERSITY

SCHOOL OF GRADUATE STUDIES

COLLAGUE OF NATURAL AND COMPUTATIONAL

SCIENCES DEPARTMENT OFENVIRONMENTAL
SCIENCES (CHEMISTRY)

M.Sc. PROPOSAL ON

ASSESSMENT OF THE LEVEL OF SELECTED PHYSICO

CHEMICALAND BACTERIOLOGICAL QUALITY OF
PARAMETERS OF WATER SUPPLY (INCASE OF NEJO
MUNICIPAL TOWN), WESTERN WOLLEGA ZONE OF
OROMIA REGIONAL STATE, ETHIOPIA.

BY: ASRESA KEBEDE EMIRU

MAIN ADVISOR: ABREHAM ABEBE (Msc)
CO-ADVISOR: MERGA ASEFA (Ms
SEMPTEMBER 2020

NEKEMTE, ETHIOPIA

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APPROVAL SHEET FOR SUBMITTING PROPOSAL

Title: assessment of the level of selected physico-chemical and bacteriologicalquality of

parameters of water supply (incase of nejo municipal town), western wollega zone of oromia
regional state, ethiopia

Submitted by: Asresa Kebede Emiru __________ _______

Name of student_______________________ signature ________ date________

approved by:

1. Abreham abebe (m.sc.) _______________

Name of major advisor signature date

2. merga asefa (m.sc.) ______________

name of co-advisor signature date

3. __________________ __________ _______

name of chairperson, dgc signature date

4. ____________________ __________ ________

name of dean, sgs signature date

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 TABLE OF CONTENTS

ABBREVIATION AND ACRONYMS.....................................................................................v

Abbreviations..............................................................................................................................v
Chapter-1.....................................................................................................................................1
1. INTRODUCTION..................................................................................................................1
1.1 Background of the study...................................................................................................1
1.2 Statement of the Problem................................................................................................2
1.3. Objective of the Study......................................................................................................3
1.3.1 General Objective..........................................................................................................................3
1.3.2 Specific Objectives........................................................................................................................3
1.4 Research Questions...........................................................................................................3
The principal research questions that are attempted to be addressed are the following.........3
1.5 Significance of the Study..................................................................................................4
1.6 Scope of the study.............................................................................................................4
2. LITERATURE REVIEW.......................................................................................................5
2.1 Definition..........................................................................................................................5
2.2 Physicochemical Quality Parameters of Drinking Water.................................................5
2.2.1 Temperature...................................................................................................................................5
2.2.2 pH...................................................................................................................................................6
2.2.3 Turbidity........................................................................................................................................6
2.2.4 Electrical Conductivity..................................................................................................................6
2.2.5 Free Residual Chlorine..................................................................................................................7
2.2.6 Total Dissolved Solids...................................................................................................................7
2.2.7 Hardness.........................................................................................................................................7
2.3 Major Chemical Parameters..............................................................................................9
2.3.1 Calcium and Magnesium...............................................................................................................9

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 2.4 Bacteriological Water Quality Parameters......................................................................12
2.4.1 Coliform Bacteria.........................................................................................................................13
2.4.2 Faecal Coliforms (Thermotolerant Bacteria)...............................................................................13
2.4.3 Faecal Streptococci and Enterococci...........................................................................................13
2.5 Water Sample Digestion..................................................................................................14
2.6 Potable Water..................................................................................................................14
2.7 Sources of Contaminants in Drinking Water..................................................................15
2.8 Health Effects of Contaminants in Drinking Water........................................................15
2.9 Standards and guide lines for drinking water quality......................................................16
Chapter-3...................................................................................................................................16
3. MATERIALS AND METHODS..........................................................................................16
3.1 Description of the Study Area.........................................................................................16
3.1.1 Study sites....................................................................................................................................16
3.1.2 Data collection techniques...........................................................................................................16
3.1.3 Research design...........................................................................................................................17
3.1.4 Sample size..................................................................................................................................17
3.1.5 Sample Collection........................................................................................................................17
3.1.6 Sample handling...........................................................................................................................18
3.2 materials..........................................................................................................................18
3.2.1 Instruments...................................................................................................................................18
3.2.2 Chemicals.....................................................................................................................................18
3.3 methods...........................................................................................................................19
3.3.1 Classical Methods of Analysis.....................................................................................................19
3.3.2 Instrumental Methods of Analysis...............................................................................................19
3.4 Sampling Procedures.......................................................................................................19
3.4.1 Cleaning of Sampling Equipment..............................................................................................19
3.4.2 Cleaning of Laboratory Glass wares............................................................................................19
3.4.3 Calibration of Analytical Balance................................................................................................19
3.5 Data Analysis..................................................................................................................19
3.5.1 Statistical Analysis.......................................................................................................................20

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3.5.2 Microbial Analyses......................................................................................................................20
Chapter-4...................................................................................................................................21
4. Work plan..............................................................................................................................21
4. 1. Budget and Time plan...................................................................................................21
Table 2. For this study the budget plan is shown in the following table..................................21
Table .3. For this study the time plan is shown in the following table.....................................21
REFERENCES.........................................................................................................................22

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ABBREVIATION AND ACRONYMS
Abbreviations D e f i n i t i o n

D A P Diammonium monohydrogen phosphat e

D O D i s s o l v e d O x y g e n

E B T E r i o c h r o m e b l a c k - T

E C E l e c t r i c a l c o n d u c t i v i t y
E D T A Ethylene diamine tetra acetic aci d

M D L M e t h o d D e t e c t i o n L i m i t
M Q L Method Quantification Limit

p H P o t e n t i a l o f H y d r o g e n

S A S Statistical Analysis Software

T A T o t a l a l k a l i n i t y

T D S T o t a l d i s s o l v e d s o l i d s

T H T o t a l h a r d n e s s

U N E P United nations environment programm e

U S E P A United states environmental protection agenc y

W H O World health organization

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 CHAPTER ONE
1. INTRODUCTION
1.1 Background of the study

Water is essential for survival of all forms of life. 80% of the earth´s surface is covered by
water. Supply of safe water is univocally a basic requirement for human consumption. Next to
oxygen water is essential for human existence (White, 1969). It is an essential nutrient, which
also sustains agriculture, allows aquatic life, supports industry, produces hydroelectricity,
permits aquatic transport, ensures personal hygiene, maintains clean environment, uses for
sport as well as for recreation (EFDR Water Resource Policy, 1997; Falkenmark, 1982).
However for most parts of the world, human existence mainly depends on fresh water supply
which is less than 1% of the water available on the earth (Reagen and Bookins-fisher, 1997).

Unsafe drinking water contributes to numerous health problems associated mainly with water
born disease (Ilkem et al., 2002). Rapid industrialization, Urbanization and indiscriminate use
of fertilizer and pesticides in agriculture are deteriorating water quality. The problems of
groundwater quality are much more acute in the areas which are densely populated and
thickly industrialized. The rapid growth of urban areas has further affected groundwater
quality due to over exploitation of resources and improper waste disposal practices. Hence,
there is always a need for and concern over the protection and management of water quality
(Patil et al., 2001).

The consequence of urbanization and industrialization leads to spoil the water. Due to use of
contaminated water human population suffer from water born disease. According to World
Health Organization (WHO), about 80% of all diseases in human beings is caused by
contaminated water (WHO, 2006). Ideally, drinking water should not contain any
microorganisms known to be pathogenic or any bacteria indicative of fecal pollution (WHO,
1993).

Therefore, monitoring the quality of water is one of the essential issues of drinking water
management (Trevett et al., 2004; Dieter and Muckter, 2007). The World Health Organization
also laid down drinking water standards which are considered as international standards. The

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water borne disease out breaks associated with the distribution system failures has been
increased over the years (Moe and Rheingans, 2006). Water has unique chemical properties
due to its polarity and hydrogen bonds, which means it is able to dissolve, adsorb or suspend
many different compounds, thus, in nature, water is not pure as it acquires contaminants from
its surrounding and those arising from humans and animals as well as other biological
activities (WHO, 2007; McMurry and Fay, 2004 ).

In Ethiopia, the dominant source of drinking water used to supply major urban and rural
communities is from wells and springs. Although there are no systematic and comprehensive
water quality assessment programs in the country, there are increasing indications of water
contamination problems in some parts of the country. The major causes of this contamination
could be soil erosion, domestic waste from urban and rural areas and industrial wastes
(Gebrekidan and Samuel, 2011).

Thus, it is needed to emphasize the quality of water in Nejo town. Nejo town is situated in the
western region ofOromia, 515 km away from Addis Ababa, the capital city of Ethiopia. The
town is one of the towns under the modern reform since 1997, so it is administered by Mayor.
It is one of the most highly developing towns of Oromia regional state and a large number of
houses are being built recently. This town is beyond the scope of Woreda inconsideration with
the number of houses and the number of people that exist in this town. The area of the town is
3291 Hectares and the number of population is about 73,000. The weather of the town is
moderately cold and the topography of the land scope is so comfortable that it’s suitable for
human life.

Even though there are no huge factories that may cause the water and soil pollution, there are
a number of small industries such as mills, dry coffee processing machines, big hotels etc. in
abundant. The only asphalt road from Addis Ababa to the western border through this town is
always busy of different kinds of vehicles transporting people and materials to the renaissance
Dam of Ethiopia. This overcrowded transportation is suspected that it may release different
chemicals that could be added to the water and will result in water pollution via a long period
of time. The surrounding farmers often use some chemicals such as fertilizers (i.e. DAP, Urea

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etc.) and weed distracting toxins which might have negative impact on the quality of the soil
and the fresh water.

The water supply office is responsible for providing the tap water supply for the population.
The source of the tap water is the pump that provides water from the underground water that
found near the town. In my study, I want to ask some questions the people of that area
regarding the back ground research about the content and quality of the water in the form of
interview. In this way, I hope that I will get some information about the knowledge of the
users of water for drinking purpose concerning the constituents, abundance and scarcity of the
selected parameters.

However regular monitoring of water quality parameters has been proposed by international
organizations like WHO. Thus it is important to assess these parameters in order to maintain
the quality of drinking water in the study area. Both standards, international and national, are
available now. These standards recommend maximum permissible limits on several water
quality parameters in order to avoid any adverse effect on the health of the population
consuming the water.

Therefore, this particular study is aimed to determine the physicochemical parameters

including temperature, pH, Electrical-conductivity, total dissolved solids (TDS), total
hardness, and major chemical parameters including SO42- ,PO43- , NO3- , Cl-, Ca, Mg, Na , K ,
Fe and Mn. It is particularly aimed to determine whether these parameters meet the WHO
(World Health Organization) and Ethiopian standards for drinking water, as well as to find out
the possible causes of any contamination to make appropriate recommendations. The results
of the study will also expect to give the baseline information for water quality study in Nejo
town local area in the future. Obviously, the quality of water is described by its physical,
chemical and biological characteristics .It is therefore important to check the water quality at a
regular interval of time.

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1.2Statement of the Problem

Water has unique chemical properties due to its polarity and hydrogen bonds which means it
is able to dissolve, absorb, adsorb or suspend many different compounds, thus, in nature,
water is not pure as it acquires contaminants from its surrounding and those arising from
humans and animals as well as other biological activities (WHO, 2007; McMurry and Fay,
2004). The international water community continues to highlight good water quality as vital
for securing the futurity of human and aquatic ecosystem health (UNEP, 2004).

However, as the researcher has tried assessing to get information regarding the quality of
water in this area no research study has been conducted. So far there is no information about
the physical as well as chemical elements of water that the dwellers are using for drinking
purpose. Thus, this is the great problem which initiates the researcher to study the quality of
the water in the study area. In addition to this idea, the amount of water supplied for the
dwellers of Nejo municipal town does not coincide with the number of people living in this
area. This means that the people is over populated (almost 73,000.00) in the study area, but
the amount of water supplied is very limited in comparison with the number of people
indicated. The over population of the people could be resulted from the migration of
surrounding small towns and villages of Nejo municipal town due to the fact that Nejo
municipal town is the town which is the commercial center for many other towns and villages
of western Wollega zone and its weather condition is very suitable for the lives of human
beings and other living organisms. On the other hand, this town is found on the main road that
goes from Finfinnee to Asosa that connects two cities of the Country.

Therefore, different vehicles pass via it and it also makes the town to be over populated from
day to day. The scarcity of water supply can be resulted from all these factors I mentioned
above. But, even though there is scarcity of water supply in the town, the researcher wants to
assess the quality of parameters of drinking water supplied for users of Nejo municipal town.
For this reason the ultimate goal of this study is to assess the current physico-chemical status
of water quality in the study area and to figure out if there is health risk associated with those
water sources by comparing with the international and national water quality standards. This

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facilitates the researcher to conduct the study in this area from the point of view of water
quality parameters.
1.3. Objective of the Study
1.3.1General Objective
The overall objective of the study is to evaluate the quality of drinking water in Nejo town
and to compare the municipal tap water with the World Health Organization quality standard
and other world water bodies’ guidelines for drinking water.
1.3.2 Specific Objectives
 To characterize the physico-chemical parameters of water on the basis of
temperature, pH, Electrical-conductivity, turbidity, color , total dissolved solids, total
hardness, SO42-, PO43-, NO3-, Cl-, Ca, Mg, Na, K, Fe, Mn
 To compare the physico-chemical and bacteriological characteristics of drinking water
of the study area with WHO guideline, Ethiopian standards, International and National
permissible limits.

 To generate a base line data on water quality status in the study area.

1.4 Research Questions

The principal research questions that are attempted to be addressed are the following.

 Do the drinking water quality parameters of Nejo town fit the guidelines set in the
WHO standard and that of the Ethiopian recommended guideline?
 If the water quality of the town, expressed in terms of the water quality
parameters, is in short of the WHO guidelines and the Ethiopian recommended
maximum and minimum permissible limits, what will be the health significance of
water on the health of users?
 What is the level of the quality of water that the dwellers of Nejo municipal town
are using for drinking purpose?

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1.5Significance of the Study

This study is expected to give the baseline information about tap water quality of Nejo town
regarding the selected physico-chemical, bacteriological and major chemical constituents. By
getting information about the present condition of the water quality pollution potential of the
naturally occurring chemicals in the study area, the Nejo town water supply office in
collaboration with Nejo town administration will take necessary remedial action. Moreover,
the study outcome will provide information about the acceptability of the water for drinking
purpose by comparing with drinking water quality Standards given by national and
international agencies.

1.6. Scope of the study

This study will be mainly carried out on the physico-chemical and bacteriological quality of
drinking water (tap water) from Nejo town on some selected parameters and the study will be
carried out in month of September , 2012.Therefore it may not explain seasonal variability of
water quality change.

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 CHAPTER-2
[

2. LITERATURE REVIEW
2.1 Definition
The search for high-quality water has been an objective of human society going back to
prehistoric times. Early humans gathered in locations with readily accessible sources of water
and if the water was believed to be of questionable quality, entire settlements would be
discarded. For improving drinking water quality paralleled studies have been established
between water born disease and water quality.

The introduction of chlorine as a chemical disinfectant was an alternative for those

communities that could not afford the expense of elaborate filtration plants. The introduction
of chlorination of drinking water was followed by a remarkable reduction in cholera,
dysentery and typhoid worldwide. Today, water treatment and specifically chlorination and/or
filtration of drinking water have been hailed as the major public health achievement of the
20th century (Kanitz et al., 1996). As the century progressed, the identification of water
contaminants shifted from microbiological to chemical. As the public health infrastructure
grew, outbreaks associated with chemical spills or leaks into potable water drew the attention
of the scientific community concern with inorganic contaminants.

2.2Physicochemical Quality Parameters of Drinking Water

Toxic chemicals in water pose the greatest threat to the safety of drinking water and their
effects are enormous which can cause damage to human health, crops and aquatic organisms
(David et al., 2011). An appreciable number of serious health concerns may occur as a result
of the chemical contamination of drinking water. The drinking water parameters (such as
temperature, pH, turbidity, EC, taste and color) play important roles in the biological and
chemical quality of drinking water (WHO, 2008).

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In the purest state, water is both odorless and tasteless: as inorganic and organic substances
dissolved in water, it begins to take on a characteristic taste and sometimes odor. Generally,
the inorganic salts at the concentration found naturally in drinking water do not adversely
affect the taste. Both taste and odors are caused by the interaction of the many substances
present. These include many particles, decaying vegetation, organisms (plankton, bacteria,
and fungi), various inorganic ions of chlorides and sulphides of sodium, calcium, iron and
manganese, organic compounds and gases. Water should be palatable rather than free from
taste and odor, but with people having very different abilities to detect taste and odors at low
concentrations this is often difficult to achieve. Offensive odors and tastes account for most
consumer complaints about water quality (Gray, 2008).

Physical and chemical properties are parameters that do not identify particular chemical
species but are used as indicators of how water quality may affect water uses. These are
temperature, hydrogen ion concentration (measured as pH), electrical conductivity, total
dissolved solids and total hardness (Weiner, 2008; USEPA, 2006).

2.2.1 Temperature
Temperature is an important biologically significant factor, which plays an important role in
the metabolic activities of the organism. Temperature is primarily an aesthetic criterion for
drinking-water. Cool water is generally more palatable than warm water, and temperature will
have an impact on the acceptability of a number of other inorganic constituents and chemical
contaminants that may affect taste. High water temperature enhances the growth of
microorganisms and may increase problems related to taste, odor, color and corrosion (WHO,
2011; Gopalkrushna, 2011).

2.2.2 pH
Measurement of pH is one of the most important and frequently used tests in water chemistry.
No health based guideline value is proposed for pH. Although pH usually has no direct impact
on consumers, it is one of the most important operational water quality parameters (WHO,
2008).pH is an important factor in determining the chemical and biological properties of

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water. It affects the chemical forms and environmental impacts of many chemical substances
in water.

For example, many metals dissolve as ions at lower pH values, precipitate as hydroxides and
oxides at higher pH, and dissolve again at very high pH. The pH value is an indicator of the
chemical state in which these compounds will be found and must be considered when
establishing water quality standards. In natural waters it usually ranges between 4.5 and 8.5.
The WHO guide level for pH in drinking water quality is 6.5 to 8.5 (ES 261, 2001; WHO,
2006). The pH of pure water at 25 oC is 7.0, but the pH of environmental waters is affected by
dissolved CO2 and exposure to minerals (WHO, 2011; Pradhan and Pirasteh, 2011; Weiner,
2008).

2.2.3Turbidity

Turbidity is a general measure of water ‘cloudiness’ created by particles suspended in a

water sample. It has been used to assess drinking water quality for a century and is still the
most widely used particle measurement in water treatment. These particles may include clay,
silt, finely divided inorganic and organic matter, soluble colored organic compounds, and
plankton and other microscopic organisms. Excessive turbidity, or cloudiness, in drinking
water is aesthetically unappealing, and may also represent a health concern as it may provide
food and shelter for pathogens. The World Health Organization (WHO, 2011) Guidelines for
drinking-water quality states turbidity in water is caused by colloidal matter or suspended
particles that obstructs light transmission through the water which may be caused by
inorganic or organic matter or a combination of the two. Microorganisms (bacteria, viruses
and protozoa) are typically attached to particulates, and removal of turbidity by filtration will
significantly reduce microbial contamination in treated water.

2.2.4Electrical Conductivity
Electrical conductivity (EC) is the measure of the ability of water to conduct an electric
current and depends upon the number of ions or charged particles in the water. EC
determinations are useful in aquatic studies because they provide a direct measurement of
dissolved ionic matter in the water. Low values are characteristic of high-quality, low-nutrient

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waters. High values of conductance can be indicative of salinity problems but also are
observed in eutrophic waterways where plant nutrients (fertilizer) are in greater abundance.
Very high values are good indicators of possible polluted sites. Conductivity often is used to
estimate the amount of TDS rather than measuring each dissolved constituent separately.
Generally the chemical composition of water determines its electrical conductivity.
(Government of Western Australia, 2009; Ethiopian Water Technology Centre; 2008; Alberta
Environment, 2000).

2.2.5 Free Residual Chlorine

Chlorine is added to drinking water supplies for the purpose of destroying or deactivating
disease causing microorganisms. This is termed as water disinfection. Chlorine is usually
added to water in liquid form as sodium hypochlorite or solid form as calcium hypochlorite.
Maintaining an adequate level of residual chlorine is of great importance in terms of
distribution system water quality management. The (WHO, 2003), guideline value for the
palatability and health significance of residual chlorine is 0.2 to 0.5mg/l in drinking water
distribution systems. The disinfection activity of chlorine on microorganisms is greatly
reduced at high pH, probably because at an alkaline pH, the predominant species of chlorine
is hypochlorite ions (Hurst, et al., 2002). If the pH of the water is high, chlorine is less
effective up on pathogens (USEPA, 1999).

2.2.6 Total Dissolved Solids

The term total dissolved solid (TDS) refer to materials that are completely dissolved in water
(WHO, 2011; Ethiopian Water Technology Centre, 2008). Determination of the “solids”
contents important for both aesthetic and practical reasons; drinking water with high solids
content can have a disagreeable palatability. The palatability of water with a TDS level of less
than about 600 mg/L is generally considered to be good; drinking-water becomes significantly
and increasingly unpalatable at TDS levels greater than about 1000 mg/L. Dissolved solids in
natural waters may consist of carbonates, bicarbonates, chlorides, sulfates, phosphates,
nitrates, magnesium, sodium, iron, manganese and other substances. Taste problems in water
often arise from the presence of high TDS levels with certain metals present, particularly iron,
copper, manganese, and zinc (Pradhan and Pirasteh, 2011; WHO, 2011; Weiner, 2008).

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2.2.7Hardness
Hardness is often referred to as the soap consuming property of water. Hardness is usually
divided into two types, carbonate and non-carbonate. Carbonate hardness usually due to the
presence of bicarbonate (Ca(HCO3)2 and Mg(HCO3)2) and carbonate (CaCO3 and MgCO3)
salts. Non carbonate hardness is contributed by salts such as calcium chloride (CaCl2),
calcium sulfate (CaSO4), magnesium sulfate (MgSO4), and magnesium chloride (MgCl2).
Total hardness equals the sum of carbonate and non-carbonate hardness. In addition to Ca ++
and Mg++, iron (Fe++), strontium (Sr++), and manganese (Mn++) may also contribute to hardness
(APHA et al., 1998). However, the contribution of these ions is usually negligible. Hardness
is usually reported as equivalents of calcium carbonate (CaCO3) and is generally classified as
soft, moderately hard, hard, and very hard. It is best to report results as the actual equivalents
of CaCO3 since the inclusive limits for each category may differ between users of the
information. The classification scheme used by the U.S. Environmental Protection Agency
(EPA) is shown in table 1

Table 1: Water hardness classifications (reported as CaCO3 equivalents) used by the U.S.
EPA (EPA 1986)
C l a s s i f i c a t i o n C a C O 3 e q u i v a l e n t
( m g / L )

S o f t < 7 5

M od er at el y h a r d 7 5 – 1 5 0

H a r d 1 5 0 – 3 0 0

V e r y h a r d > 3 0 0

Hardness caused by Ca and Mg is usually indicated by precipitation of soap scum and the
need for excess use of soap to achieve cleaning. The taste threshold for the Ca ion is in the
range of 100 – 300 mg/L, depending on the associated anion, and the taste threshold for Mg is
probably lower than that for Ca. In some instances, consumers tolerate water hardness in
excess of 500 mg/L (WHO, 2011; Pradhan and Pirasteh, 2011; Ethiopian Water Technology

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Centre, 2008). As with alkalinity, hardness is expressed as an equivalent concentration of
CaCO3. However, hardness is a property of cat ions (Ca2+ and Mg2+) while alkalinity is a
property of anions (HCO3-and CO32-) (Weiner, 2008).

There are two methods used in determining the hardness of water. The one described and used
here in this study is based on a titration method making use of a chelating agent. The basis for
this technique is that at specific pH values, ethylene diaminetetra acetic acid (EDTA) binds
with divalent cat ions to form a strong complex. Thus, by titrating a sample of known volume
with a standardized (known) solution of EDTA, one can measure the amount of divalent
metals in solution. The titration’s endpoint is observed using a colorimetric indicator,
Eriochrome Black T. When a small amount of Eriochrome Black T (blue in color while in the
absence of divalent metals) is added to a solution containing hardness (at pH = 10.0), it
combines with a few of the hardness ions to form a weak wine red complex. During the
titration with EDTA, more and more of the hardness ions are complexed with EDTA.
Eventually, there is a point at which the EDTA has complexed all of the free ions and “out-
competes” the weaker Eriochrome Black T complex for hardness ions. At this point, the
Eriochrome Black T returns to its uncomplexed blue color, indicating that only EDTA-
complexed hardness ions are present in the endpoint of the titration

2.3Major Chemical Parameters

Major chemical parameters are those most often present in natural waters in concentrations
greater than 1.0 mg/L. These are the cat ions Ca 2+, Mg2+ and the anions Cl- , NO3-and SO42-
(Weiner, 2008; USEPA, 2006).

2.3.1Calcium and Magnesium

2.3.1.1 Calcium (Ca)
Ca is an essential nutrient for plants and animals, essential for bone, teeth, blood coagulation,
nervous system, and cell development. Recommended daily intakes for adults are between
800 and 1200 mg/day. Most of this is obtained in food; drinking water typically accounts for
50–300 mg/day, depending on the water hardness and assuming ingestion of 2 L/day. Ca in
food and water is essentially nontoxic. A number of studies suggest that water hardness

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protects against cardiovascular disease. One possible adverse effect from ingestion of high
concentrations of Ca for long periods of time may be a greater risk of kidney stones. The
presence of Ca in water decreases the toxicity of many metals to aquatic life. Thus, the
presence of Ca in water is beneficial and no limits on Ca have been established for protection
of human or aquatic health (Weiner, 2008; USEPA, 2006).

Calcium ions are contained in almost all spring drinking water. Health effects caused by hard
water, very rich in calcium and magnesium, are unknown (Josette, 2000). An excess in
calcium can alter the water taste or cause scaling problems in pipes and household appliances.
In the reduction of the content of calcium and magnesium ions dissolved in water it is
recommended that the calcium content never goes under 60 mg/L. The World Health
Organization has recommended a minimum calcium daily intake of about 700 mg. Drinking
calcium poor water is considered dangerous for the risk of coronary diseases. Ca is
determined by EDTA titration method and calculated as Calcium hardness (mg/L as CaCO3).

2.3.1.2 Magnesium (Mg)

Mg is an essential nutrient for plants and animals, essential for bone and cell development.
Recommended daily intake for adults is 400–450 mg/day, of which drinking water can supply
from 12 to 250 mg/day, depending on the Mg concentration and assuming ingestion of 2
L/day. In general, the presence of Mg in water is beneficial and no limits on Mg have been
established for protection of human or aquatic health (Weiner, 2008). There are no primary or
secondary drinking water standards for Mg. Mg in drinking water may provide nutritional
benefits for persons with Mg deficient diets (USEPA, 2006). It is, with sodium and calcium,
among the cat ions most commonly found in drinking water. In humans magnesium is
important for many metabolic functions and for muscular and nervous activity (Williams,
1998). The daily recommended intake is 150-500 mg. Mg in water sample can be calculated
from the Ca hardness and TH.

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2.3.1.3 Sodium (Na)

Na will be analyzed by the validated analytical methods for the required analytes and the
obtained results will be given in table. The concentration of Na in water sample will be
calculated from calibration carve of the standard solution prepared in the procedure using the
Regression equation (calibration equation) of the calibration curve.

2.3.1.4 Potassium (K)

The concentration of K in water sample will be calculated using calibration equation of the
calibration carve of the standard solution prepared through the entire procedure.

2.3.1.5 Iron and manganese

Iron and manganese can be naturally found in groundwater and some surface water (such as
creeks, rivers and some shallow dug wells) (Rosen and White, 2001), these metals from
natural source usually occur together, therefore monitoring for one could be used as an
indicator of possible problems with both parameters (Howard et al., 2003). However, human
activities may also be responsible for manganese contamination in water in some areas. There
are areas of the world that have naturally high amounts of iron in their groundwater. Iron can
also be found in drinking water that is passed through rusty steel or cast iron pipes (McNeill
and Edwards, 2001).

Iron and manganese can come in two forms in water: dissolved and suspended. If
groundwater comes from a deep tube well, the iron and manganese may be dissolved and not
visible. However, once the iron is exposed to air, it usually turns the water black or orange
colour. In surface water, manganese can be dissolved or suspended. If surface water has iron
in it, it will be a red-orange colour from the iron that is suspended in the water. Water with
high levels of suspended manganese usually has a black colour or black flakes in it. Drinking
water with high concentrations of iron and manganese may be present in water, are likely to
affect the acceptability of water for drinking, but have limited direct health significance.

Iron and manganese, however, can change the colour of water and it may cause people to not
use it and choose another, possibly contaminated, water source instead (Howard et al., 2003).
People need small amounts of manganese to keep healthy and food is the major source for

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people. However, too little or too much manganese can cause adverse health effects (Ljung
and Vahter, 2007)

2.3.1.6 Chlorides

Chlorides are widely distributed in nature, usually in the form of Na, K, and Ca salts (NaCl,
KCl and CaCl2), although many minerals contain small amounts of chloride as an impurity.
Chloride in natural waters arises from weathering of chloride minerals, salting of roads for
snow and ice control, seawater intrusion in coastal regions, irrigation drainage, ancient
groundwater brines, geothermal waters, and industrial wastewater. Concentrations in
unpolluted surface waters and non-geothermal ground waters are generally low, usually below
100 mg/L. Thus, chloride concentrations in the absence of pollution are normally less than
those of sulfate or bicarbonate. Chloride is the most abundant anion in the human body and is
essential to normal electrolyte balance of body fluids. Chlorine is effective disinfectant and
very easy to apply either as a gas or as the hypochlorite (calcium hypochlorite powder, or
sodium hypochlorite liquid) in controlled pH condition. In water chlorine reacts to form
hypochlorous acid (HOCl), a very effective disinfectant. The hypochlorous acid dissociates to
form hypochlorite ion (OCl-) which is estimated to be 150 to 300 times less effective as a
disinfectant than hypochlorous acid.

The relative proportions of these two compounds in solution depends on pH and to a lesser
extent on temperature, with lower pH and temperature resulting in higher proportions of
HOCl results in the disinfection process (Metcalf and Eddy, 2003). Chlorides in water are
more of a taste than a health concern, although high concentrations may be harmful to people
with heart or kidney problems. There are no primary drinking water standards for chloride.
The EPA secondary standard for chloride is 250 mg/L, based on adverse effect on taste
(WHO, 2011; Pradhan and Pirasteh, 2011; Weiner, 2008). Chloride in sample water is
determined by Argentometric titration.

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2.3.1.7 Fluoride

The WHO suggests that drinking water should have 0.5 – 1.0 mg/L fluoride to protect teeth.
Many cities around the world add fluoride to their drinking water to reach this level. Higher
amounts of fluoride between 1.5 – 4.0 mg/L can cause dental fluorosis. Very high amounts of
fluoride greater than 10.0 mg/L can lead to skeletal fluorosis. This is why the WHO suggests
that drinking water should not have more than 1.5 mg/L of fluoride. Currently research is
being conducted to develop HWT technologies that can effectively remove fluoride from
drinking water. In the meantime, household that use drinking water with concentrations
greater than 1.5 mg/L should try to use an alternative water source if at all possible

Fluoride can naturally occur in groundwater and some surface water. Drinking water is
normally the major source of fluoride exposure (Kaseva, 2006), with exposure from diet and
from burning high fluoride coal also major contributors in some regions (Ando et al., 1998).
High levels of fluoride can be found naturally in many areas of the world including, Africa,
the Eastern Mediterranean and southern Asia. One of the best known high fluoride areas
extends from Turkey through Iraq, Iran, Afghanistan, India, northern Thailand and China
(Petersen et al., 2005). However, there are many other areas with water sources that contain
high fluoride levels and which pose a risk to those drinking the water, notably parts of the rift
valley in Africa.

2.3.1.8Nitrates

Nitrates are the most oxidized forms of nitrogen and the end product of the aerobic
decomposition of organic nitrogenous matter. High nitrate concentrations in domestic water
supplies can be toxic to human life. The significant sources of nitrates are chemical fertilizers
from cultivated lands, drainage from livestock feeds, as well as domestic and industrial
sources. In soil, fertilizers containing inorganic nitrogen and wastes containing organic
nitrogen are first decomposed to give ammonia, which is then oxidized to nitrite and nitrate.
High concentrations of nitrate in surface or groundwater generally indicate agricultural
contamination from fertilizers and manure seepage (California Department of Public Health,
2012; WHO, 2011).

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Natural waters in their unpolluted state contain only minute quantities of nitrates. Drinking
water standards for nitrate are strict because the nitrates can be reduced to nitrites, which
oxidize iron in blood hemoglobin from ferrous iron (Fe 2+) to ferric iron (Fe3+). The resulting
compound, called methemoglobin, cannot carry oxygen. The resulting oxygen deficiency is
called methemoglobinemia or blue baby syndrome which is caused by high nitrate levels in
water and this condition found especially in infants less than six months. The stomach acid of
an infant is not as strong as in older children and adults. This causes an increase in bacteria
that can readily convert nitrate to nitrite (NO 2-). Methemoglobin does not carry oxygen
efficiently.this results in a reduced oxygen supply to vital tissues such as the brain.
Methemoglobin in infant blood cannot change back to hemoglobin, which normally occurs in
adults. Severe methemoglobinemia can result in brain damage and death (Somaini and
Quirindongo, 2004). A health-based guideline value for nitrate of 50 mg/L (expressed as
nitrate ion) was recommended in the WHO Guidelines for drinking water quality to prevent
methemoglobinemia (WHO, 2011).

2.3.1.9Sulphates

Sulfate (SO42-) is found almost universally in natural waters at concentrations ranging from a
fewtenths to several thousand milligrams/liter. The highest concentrations are usually found
in groundwater and are considered to be a mixture of SO 42- from atmospheric, geochemical,
and biological sources. Approximately 30% of SO42- in groundwater may be of atmospheric
origin, and the remainder from geologic and biological processes. SO42- ions are discharged
into surface water through industrial wastes and atmospheric deposition of SO 2 (USEPA,
2008). The presence of SO42- in drinking water can cause noticeable taste. It is generally
considered that taste impairment is minimal at levels < 250 mg/L (Pradhan and Pirasteh,
2011; WHO, 2011).Sulphates are sulphuric acid salts combined with metallic ions. Water can
naturally contains small quantities of sulfates, but they are mostly transferred in water bodies
from the atmosphere and in the atmosphere from road traffic, industries an energetic
production. Sulphur oxidized in the air can come back on the soil as acid rain causing serious
environmental problems. Sulfate is a substance that occurs naturally in drinking water. Health

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concerns regarding sulfate in drinking water have been raised because of reports of diarrhea
associated with the ingestion of water containing high levels of sulfate (USEPA, 1999).

2.3.1.10. Phosphate (PO43-)

Water may contain PO43- derived from natural contact with minerals orthrough pollution from
application of fertilizer, sewage and industrial waste. Groundwater,therefore, is more likely to
have higher PO43- concentration (Maiti, 2004). The PO43- in the water samples may be
possibly from the natural contact with minerals when the water bodiescome across the layers
of various minerals. The concentration ofPhosphate in water sample will also be calculated
from calibration equation of the calibration curve of the standard solution.

2.4 Bacteriological Water Quality Parameters

The presence of certain microorganisms in water is used as an indicator of possible
contamination and an index of water quality (Hurst et al., 2002). Indicator organisms are
selected to demonstrate the presence of human and animal wastes and hence the potential
presence of pathogens in drinking water. Indicator organisms are usually of intestinal origin
from humans and animals (Muyima and Ngcakani, 1998; Hurst et al., 2002; Brian,

2002; Mombal, et al., 2006; Plummer et al., 2007).

Therefore, the presence of indicator organisms in water indicates contamination of water by

faecal matter, which could probably contain pathogens such as Salmonella and Shigella
(Gundry et al., 2004). According to USEPA regulations three main groups of bacteria are
suggested to serve as indicators to monitor water quality. These are total coliforms (TC),
faecal coliforms (FC), and enterococcus (USEPA, 1998).

The criteria set to identify indicator organisms for water quality analyses are: the organisms
must be exclusively of faecal origin and consistently present in fresh faecal waste, they must
occur in greater numbers than the associated pathogens, they must be more resistant to
environmental stresses and persist for a greater length of time than the pathogen, they must
not proliferate to any greater extent in the environment or they should not grow in natural

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waters, and they have to be detected on the basis of simple, reliable, and inexpensive
methods(Hurst et al., 2002;WHO, 2004).

2.4.1 Coliform Bacteria

Total coliforms are the ones that are commonly measured as indicator bacteria for drinking
water quality (Brian, 2002; Hurst et al., 2002). They are defined as aerobic and facultatively
anaerobic non spore forming bacteria that ferment lactose at 35 to 370C with the production
of acid and gas within 24-48 hours (WHO, 1985; Hurst et al., 2002). Coliform bacteria
belong to the family enterobacteriace and include Escherichia coli (E.coli) as well as various
members of the genera Nitrobacteria, Klebsiella and Citrobacter (Hurst et al., 2002).These
bacteria originate in the intestinal tract of warm blooded animals and can be found in their
wastes. They can also be found in soil and on vegetation (Brian, 2002; Nold, 2008). Although
coliform bacteria are not pathogens, their presence indicates the possibility of finding
pathogens in drinking water (Nold, 2008). Consequently, they are used to assess possible
faecal contamination or water pollution from sewage.

According to Hurst et al (2002), the persistence of total coliform bacteria in aquatic systems is
comparable to that of some of the waterborne bacterial pathogens. Furthermore, coliform
bacteria are relatively simple to identify and are present in much larger numbers than more
dangerous pathogens (Brain, 2002; Hurst et al., 2002). For this reason the degree of faecal
pollution and the presumed existence of pathogens can be estimated by monitoring coliform
bacteria (Volk et al., 2002).

2.4.2 Faecal Coliforms (Thermotolerant Bacteria)

Thermotolerant bacteria are found in the subgroup of coliform bacteria that grow at 44°C
(Aliev et al., 2006). Faecal coliforms live in the intestines’ of warm blooded animals (Garcia-
Armisen and Servais, 2006; Howarth, 1996). As a result, they show excellent positive
correlation with faecal contamination of water from warm blooded animals (Hurst et al.,
2002). Apart from the fact that the faecal coliform E.coli is considered as one indicator of
faecal contamination of water (Stephen and Gundry, 2004), some strains such as

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enterhemorrhagic and entroinvasive have become serious causative agents of emerging
waterborne diarrhoeal disease (Nold, 2008). The presence of coliform bacteria in potable
water indicates unsuitable sanitation practices (Howarth, 1996; Garcia-Armisen and Servais,
2006). Such occurrences may be a result of poor water treatment systems, leakages in the
pipelines, and or regrowth in the distribution system (Geldreich, 1996).

2.4.3 Faecal Streptococci and Enterococci

Faecal streptococci and enterococci are gram positive bacteria (Howarth, 1996; Hurst et el,
2002; McDonald et al, 2008). Faecal streptococci are excreted by all warm-blooded animals
(WHO, 2003). The presence of faecal streptococci and enterococci in water is an indicator of
faecal pollution of water and the possible presence of enteric pathogens in water (WHO,
2003). They are widespread in the environment, wherever there is high human and animal
activity (WHO, 2004). For this reason the ratio of FC/FS indicates the origin of faecal
contamination whether it is derived from animals or human beings (Howarth, 1996). In
addition to serving as indicator bacterium, some species of streptococci has an interesting
history of wide spread distribution to have causing different diseases with more morbidity in
humans over the centuries than almost any other bacteria (WHO, 2004).

Faecal streptococci and enterococci have received widespread acceptance as useful indicators
of microbiological water quality, because; they show a high and close relationship with health
hazards, mainly gastrointestinal symptoms, associated with bathing in aquatic environments,
they are not ubiquitous as coliforms, they are always present in the faces of warm-blooded
animals, and their die-off is less rapid than that of coliforms in water and their persistence
patterns are similar to those of potential water borne pathogenic bacteria(Hurst et al., 2002)

2.5Water Sample Digestion

Samples are acidified with HNO3/HCl or with HNO3 and HCl and heated on a hotplate to
reduce the volume to a defined level. After filtration, the samples are ready for analysis
(Maiti, 2004). The relatively clean water sample such as groundwater or the tap water sample
does not require acid digestion (Zhang, 2007).

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2.6. Potable Water

Good-quality drinking water may be consumed in any desired amount without adverse effect
on health. Such water is called ‘potable’. It is free from harmful levels of impurities such as
bacteria, viruses, minerals, and organic substances. It is also aesthetically acceptable and is
free of unpleasant impurities, such as objectionable taste, color, turbidity, and odor. As fresh
water supplies are further stretched to meet the demands of industry, agriculture and an ever-
expanding population, the shortage of safe and accessible drinking water will become a major
challenge in many parts of the world and there is a growing concern for the safety and quality
of drinking water. The most common problems in household water supplies may be attributed
to hardness, iron, sulfides, sodium chloride, acidity, and disease-producing pathogens, such as
bacteria and viruses (Mahmoud et al., 2001).

The purity of drinking water also depends on the sources and water type. The water used may
be from any source, including spring water, well purified water, municipal water, or even
untreated or contaminated water (Calderon,2000). Because of the large number of possible
hazards in drinking water, the development of standards for drinking water requires
significant resources and expertise, which many countries are unable to afford. Fortunately,
guidance is available at the international level. The World Health Organization (WHO)
publishes guidelines for drinking-water quality which many countries use as the basis to
establish their own national standards. The guidelines represent a scientific assessment of the
risks to health from biological and chemical constituents of drinking water and of the
effectiveness of associated control measures. When adapting the guideline values to national
standards WHO recommends that social, economic and environmental factors are taken into
account through a risk-benefit approach. Many countries regulate the quality of water through
government standards, typically used to ensure that water quality is safe (UN World Water
Development Report, 2006).

2.7Sources of Contaminants in Drinking Water

There is no such thing as naturally pure water. In nature, all water contains some impurities.
As water flows in streams, sits in lakes, and filters through layers of soil and rock in the

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ground, it dissolves or absorbs the substances that it touches. Some of these substances are
harmless. In fact, some people prefer mineral water precisely because minerals give it an
appealing taste. However, at certain levels of minerals, just like manmade chemicals, are
considered contaminants that can make water unpleasant or even unsafe. A number of
chemical contaminants have been identified in drinking water.

The chemical contaminants for which epidemiologic studies have suggested a risk associated
with their presence in potable water include: aluminum, arsenic, disinfection by-products
(DBPs), fluoride, lead, nitrate, pesticides, cadmium, mercury and sulfate. The contaminants
are of both inorganic and organic origin. The source of the contaminant can be from point and
non-point sources of pollution, naturally occurring, come from the treatment process or
through materials used in distribution systems (Calderon, 2007). Naturally occurring
contaminants are generally the result of leaching from geologic formations and are found
primarily in groundwater. Ranges of concentrations of these contaminants range from less
than Nano-grams per liter (ng/L) to milligrams per liter (mg/L). Point sources of drinking
water contaminants include direct dumping of chemicals from domestic and industrial
sewage.

2.8 Health Effects of Contaminants in Drinking Water

The quality of drinking water is of growing concern because of the microbial contamination
and hazardous levels of chemicals that are found in drinking water in many places throughout
the world (WHO and UNICEF, 2006). Determining the health effects of these contaminants is
difficult, especially since researchers are still learning how chemicals react in the body to
damage cells and cause illness (Calderon, 2007).

As mentioned previously, concern with chemicals in drinking water started in outbreak

situations where individuals became acutely ill. Chemical spills or leaks still occur causing
acute like toxicity (primarily vomiting). As more chemicals could be found in potable water,
studies began to appear in the literature linking health effects with occurrence of the
contaminant of interest. Cancer has been one of the most popular endpoints to study in
relationship to effects associated with exposure to specific chemicals in water. Recent years
have seen an interest in reproductive and developmental effects. Studies of cancer and

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reproductive effects have been aided by the existence in many communities of databases of
mortality or morbidity for these endpoints. The epidemiologic evidence in conjunction with
toxicological data (human and animal) has been considered important in establishing causal
relationships between the exposure and effects for arsenic, lead, nitrate, radon, etc. (Mahmoud
et al., 2001) .

Toxic doses of chemicals cause either acute or chronic health effects. An acute effect usually
follows a large dose of a chemical and occurs almost immediately. Examples of acute health
effects are nausea, lung irritation, skin rash, vomiting, dizziness and even death. The levels of
chemicals in drinking water, however, are seldom high enough to cause acute health effects.
They are most likely to cause chronic health effects –effects that occur after long exposure to
small amounts of a chemical. Examples of chronic health effects include cancer, birth defects,
organ damage, disorders of the nervous system, and damage to the immune system. Evidence
relating chronic health effects to specific drinking water contaminants is limited. In the
absence of exact scientific information, scientists predict the likely adverse effects of
chemicals in drinking water using laboratory animal studies and, when available, human data
from clinical reports and epidemiological studies (Gleick, 2004)

2.9Standards and guide lines for drinking water quality

A drinking water quality guideline value represents the concentration of a constituent that
does not result in any significant health risk to the consumer over a lifetime of consumption
which describes the quality parameters set for drinking water (WHO, 2006). Drinking water
should be suitable for human consumption and for all usual domestic purposes. When a
guideline value is exceeded, the cause should be investigated and corrective action taken. The
WHO produce international on water quality and human health in the form of guide lines that
are used as the base for regulation and standard setting for those countries without legislative
administrative frame work. The guide line describe reasonable minimum requirements of state
practice to protect the health of consumer and/or derive numerical guide line values for
constituent of water or indicators of water quality (WHO, 2006).

However, neither minimum state practice nor the numerical guide line values are mandatory
limits. In order to define such limits, it is necessary to consider the guide line in the context of

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local or national environmental, social, economic and cultural condition (Ethiopian Water
Technology Centre, 2008). The WHO and different countries have given the significant
concentration with respect to several chemicals that might be present in drinking water
(appendix III). Above this level such chemical can cause undesirable effect (Ethiopian Water
Technology Centre, 2008).

CHAPTER THREE
3. MATERIALS AND METHODS
3.1 Description of the Study Area
3.1.1 Study sites

Nejo town covers an area of about 3291 Hectaresin the western Wollega which is situated in
9º 30’0” N, 35º 30’0” E. The town is located at an altitude of about 1949 m above the sea
level. The average rainfall is 1350 mm/year. The minimum and maximum annual average
temperatures are 12.2oc and 35oc respectively. The total number of population is about
73,000.Almost the total rainfall occurs during the rainy season. The study will be carried out
in samples of municipal water (tap water) collected from four major sites of Nejo town.
Details of study locations and sources of water will be given in table.

3.1.2 Data collection techniques

The research will be conducted through both primary and secondary data collections. The
primary data will be obtained from field observation, checklists, questionnaires and laboratory
results Secondary data will be collected from published and unpublished literatures and from
the sub-city reports.

3.1.3 Research design

The study will be carried out to assess the physico – chemical and bacteriological quality of
parameters of water from main source (MS), Reservoir, public stand pipeline (main

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distribution to private pipeline (bono). The study will be conducted in Nejo municipal town,
from May 2012 to October 29, 2013. The study uses laboratory analysis (experimental facts)
to estimate the level of water quality parameters of tap water that the dwellers of Nejo
municipal town are using at the moment.

3.1.4 Sample size

Total number of customers in the town for 2012 is 73,000; from these beneficiary customers,
sample population will be determined by using the following statistical formula (Yemane,
1967).

n = 𝑵/1+N x (𝒆2), Where: n= is the sample size,

N= is the population size, and
e= is the level of precision (5%)
n = 𝑵/1+N x (𝒆2), n = 21,900/1+21,900(5%), n=393

List of kebeles Total household Sampled HH

Nejo -01 5535 99
Nejo -02 5625 101
Nejo -03 5430 97
Nejo -04 5310 95
Total 21900 392

3.1.5 Sample Collection

The method of sample collection will be according to WHO drinking water guideline (WHO,
2006), OXFAM-Delagua potable water testing kit (OXFAM, 2004) and American public
health association guideline (APHA, 1998). For the bacteriological analysis, water samples
will be collected in sterile glass bottles and transported to the laboratory in ice box containing
ice freezer packs.
From each sampling point, 300ml samples will be taken for analyses. For the chlorinated
water samples sodium thiosulphate will be added to stop the chlorination process during
transportation. Three milliliter (3ml) sodium thiosulphate will be added into each sampling
bottle and sterilized in the autoclave for 15 minutes at about 121ºC.

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The bacteriological testes will be undertaken within 6 hours after collection to avoid the
growth or death of microorganisms in the sample (WHO, 2006). With regard to the
physicochemical analysis, 200ml glass water samples will be collected, labeled and
transported to Wollega University and Nekemte municipal town water supply office for
microbiology laboratory in ice box.
3.1.6 Sample handling

3.1.6.1. Sampling time, storage and transportation

The representative water samples will be taken during spring in september,2012E.C. The
water Samples will be collected at different intervals directly from the mentioned sources in
pre-cleaned, acid washed, polyethylene plastic bottles of a litter capacity. The sampling
bottles will be labeled and stored in an ice-box blow a temperature of 4 and transported to the
Nekemte Municipal water supply office for laboratory analysisof Physico-chemical
parameters such as water temperature, pH, conductivity, total dissolved solids, total hardness,
SO42-, NO3-, Cl- , Ca, Mg using standard methods. The description of sampling sites will be
given in table.

3.2 materials
3.2.1 Instruments
Instruments Polyethylene bottles, Ice-box, a mercury thermometer (0-100 0C), a pH-meter
(ELMEIRON® Zabrze-Grzybowice, POLAN), a hot plate (Corning®Hotware, PC-
200/210/220), a 0.45-μm type membrane filter paper (Whatman®, No. 41, England), Electric
Blast Drier (101-0, Tianjin Taisite Instrument Co. Ltd), electronic analytical balance with
±0.00001g precision (AA-200DS, Denver Instrument Company),
Conductivity/TDS/Salinity/Resistivity Meter (SCHOTT-GERATE GmbH. Hattenbergstr.10,
SX713, TDS working range: 0-100 g/L), Electrical Thermostatic Water Baths (DK-2000-
IIIL,Tianjin Taisite Instrument Co.Ltd), ESCO FrontierTM laboratory fume hood (Model:
EFH-4A1), ELICO-CL, 378 Flame Photometer, and ELICO-SL 160 Double Beam UV-Vis
Spectrophotometer will be used.

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3.2.2 Chemicals
HNO3 (65.0−68.0%, UNI-CHEM® Chemical reagents, India), NaCl (99.80%, UNI-CHEM®
Chemical reagents, India), KCl, KNO3 (99%, General Purpose Reagent, BDH Chemicals Ltd
Poole England) and anhydrous extra pure K2HPO4 (98-99%, KIRAN LIGHT
LABORATORIES™), Disodium salt of EDTA (99%, UNI-CHEM® Chemical reagents,
India), EBT (99.5%, UNICHEM® Chemical reagents, India), and CaCO 3 (96%, FINKEM,
Analytical grade crystals), Monohydrate manganous sulfate (MnS0 4.H2O), Potassium iodide
(KI), Conc. H2S04 ,Sodium thiosulfate (Na2S2O3.5H2O), Sodium hydroxide (NaOH), Sodium
azide (NaN3), Starch powder(indicator), H2SO4 (96.0-98.0%, UNI-CHEM® Chemical
reagents, India), AgNO3 (99%, LOBA CHEMIE PVT.LTD.INDIA) and BaCl2 (UNI-CHEM®
Chemical reagents, India) will be used without prior purification. Distilled water will be used
for all preparations and dilutions of solution
3.3 methods
3.3.1 Classical Methods of Analysis
All parameters including total hardness (EDTA Titrimetric Method), calcium hardness
(EDTA Titrimetric Method), magnesium (Calculation Method), sulphate (Turbidimetric
Method) and chloride (Argentometric Method)will be analyzed following the handbook of
methods in environmental studies (Maiti, 2004) and the standard method (APHA, 1999).

3.3.2 Instrumental Methods of Analysis

UV-Vis spectrophotometers (for SO42- and NO3- analysis) will be used.

3.4. Sampling Procedures

3.4.1. Cleaning of Sampling Equipment
All polyethylene bottles used for water sampling will be thoroughly washed with tap water
and detergent, rinsed with distilled water and then will be soaked with 10% (v/v) HNO 3. All
polyethylene bottles used for water sampling for phosphate analysis will be thoroughly
washed with tap water and will be well rinsed with chromic acid followed by distilled water,
and then it will be soaked with 10% (v/v) HNO 3. After that, all containers will be well-rinsed
with distilled water routinely; and air will be dried and stored with the caps on to prevent

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contamination. The containers will be rinsed with sample water prior to actual sample
collection before being used for sampling (APHA, 1999).

3.4.2 Cleaning of Laboratory Glass wares

All the glass wares and apparatus used through the entire analysis will be first washed with
tap water anddetergent. Next, it will be rinsed with distilled water and followed by 10% (v/v)
HNO3 solution. Finally, it will be rinsed again with distilled water and the air will be dried to
ensure that it is free from contamination.
3.4.3. Calibration of Analytical Balance
AA-200DS Model electronic analytical balance will be calibrated first properly using a known
100 g calibration weight.
3.5. Data Analysis
The data collected will be analyzed using Microsoft Excel. The laboratory results of water
quality will be summarized and compared with the National and international (WHO) guide
lines of water quality standards. Tables and graphs will be used to display the results of the
study.
3.5.1. Statistical Analysis
Microsoft Excel and SPSS (version 20) will be used to analyze the data. Descriptive statistics
like percentage, mean, standard deviation and range will be used to describe the findings. The
comparison between variables will be performed at 5% level of significance. Significance
level will also be used to indicate the associations between parameters (Naing, 2006). Molla
and Steffan (2006) as quality control method the upper and lower control limits of each
sampling site parameters will be done. The Upper and Lower control limits are lines drawn
+/- three standard deviation of the data above and below the mean value of the data set,
respectively. Each results of water quality parameters falling within these two control limits
either crossing the WHO limit or not, will be called safe from polluting the existing water
bodies. However, others falling outside the limits and crosses the WHO limits have high risk
of water quality.
To calculate the control limits, first priority will be given to find the Mean and standard
deviation of each parameter and then the upper and lower control limits will be calculated
UCL= mean +3 (s.d/n1/2) .………………………… (1)

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LCL= mean -3 (s.d/n1/2) ………………………….. (2)
Where s.d= standard deviation n = no of sampling (triplicate)
Equation (1) in the above defines the upper control limit (UCL), which is a horizontal line in
the quality control graph and lie at three times standard deviation above the mean. Whereas,
equation (2) explains the lower control limit (LCL), which is also a horizontal line in the
quality control graph and lies three times standard deviation below the mean.
3.5.2 Microbial Analyses
Bacteriological analyses for total coliform, fecal coliform and E-coli will be done using
membrane filtration (MF) according to (APHA, 1998; WHO, 2006 ;). McConkey agar and
Plate count agar media will be prepared and autoclaved at 1210C for 15 minutes at 15Ib
before being inoculated with membrane filters (Anonymous, 1982; APHA, 1992).The results
will be expressed as colony forming units (CFU) per unit volume i.e number of colonies per
100 mL of sample using the following formula No. of colonies per 100 mL = [(No. of
colonies) / (volume filtered)] × 100

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 Chapter-4
4. Work plan
4. 1. Budget and Time plan
For successful achievement of our work and to have organized plan, clear identification of
time and budget is very important. Accordingly, the following budget and time plan is
prepared.
Table 2. For this study the budget plan is shown in the following table
N I t e m Q u a n t i t Unity cost Total
o s y cost(ETB)
1 Duplicating paper 5 2 5 1 0 0
r e a m s 0 0
2 P e 5 1 5 7 5
n
3 Duplicating paper 1 5 0 5 0
4 F l e s 2 3 0 6 0
h 0 0
5 Editing and printing 7 0 5 3 5 0
0 0
6 P h o t o 1 0 0 1 1 0 0
c o p y 0 0
7 Using software 2 0 0
0
8 T e l e p h o n 6 0
e 0
9 I n t e r n e 5 0
t 0
1 0 Transportation 2 0 0
0
1 1 Analyzing 2 0 , 0 0
chemicals 0
1 2 C o m p u t e 1 5 0 0
r 0
1 3 Contingency allowance 4 0 0
0
1 4 T o t a 5 0 3 2
l 5

xxxvi | P a g e
Table.3. For this study the time plan is shown in the following table

January

March

May
April

June

August
September

October

November

February

July
N A c t i v i t i e
o s

1 Problem X
identification

2 Submission of research X
title

4 Proposal X
submission

5 Defense of X
proposal

6 Collecting X
sample

7 S a m p l e X
a n a l y s i s

8 Writing the first draft of X

analysis

9 Writing final X
thesis

1 0 Submission of the final X

thesis

xxxvii | P a g e
 11 D e f e n s X
e

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