EFFECT OF THE COLOUR OF CONTAINER AND STORAGE CONDITION

ON BACTERIOLOGICAL QUALITY OF BOREHOLE WATER IN OWO

TUGBIYELE OLAYINKA ELIZABETH

SO4/SLT/2015/2089

A PROJECT SUBMITTED TO THE DEPARTMENT OF SCIENCE

LABORATORY TECHNOLOGY, FACULTY OF APPLIED SCIENCES, RUFUS
GIWA POLYTECHNIC OWO, ONDO STATE

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD

OF HIGHER NATIONAL DIPLOMA (HND) IN SCIENCE LABORATORY
TECHNOLOGY

APRIL, 2022.

8
 CERTIFICATION

This is to certify that this project work was written and submitted by

TUGBIYELE OLAYINKA ELIZABETH with Matriculation Number

SO4/SLT/2015/2089 to the Department of Science Laboratory Technology in partial

fulfillment of the requirements for the award of Higher National Diploma (HND) in

Department of Science Laboratory Technology, Faculty of Applied Sciences, Rufus

Giwa Polytechnic, Owo, Ondo State.

_____________________ ___________
MR. OMONIYI V. O. Date
PROJECTSUPERVISOR

_____________________ ___________
MR. ADEDAPO E. B. Date
HEAD OF DEPARTMENT

2
 DEDICATION

This project is dedicated to Almighty God, the Alpha and the Omega who has

been my God from the very beginning till the present moment.

8
 ACKNOWLEDGEMENT

First and foremost, I give thanks to the Almighty God for His guidance and

protection over me throughout the duration of my programme.

The success of this work hinges on the efforts of many people. My appreciation

goes to my supervisor Mr. Omoniyi V. O. for his tireless work, interest and support, and

his useful and insightful assistance throughout the study.

My special appreciation goes to the HOD and all lecturers in the Department of

Science Laboratory Technology for their support throughout my studies in the Polytechnic.

My gratitude goes to my sweet Mr. & Mrs. Tugbiyele for their compassion, prayers

and supports rendered to me ever since my childhood till they departed from this sinful

world,

To my wonderful siblings; Taiwo and Olaoluwa God bless you all for being there at

the point of needs and I wish us all success in all our endeavours.

I will like to appreciate my friends both in the school and outside for your kindness

and supports.

God bless u all.

4
 TABLE OF CONTENTS

TITLE PAGE i

CERTIFICATION ii

DEDICATION iii

ACKNOWLEDGEMENT iv

TABLE OF CONTENTS v

LIST OF TABLES ix

ABSTRACT x

CHAPTER ONE 1

1.0 INTRODUCTION 1

1.1 Research Problem 2

1.2 Justification of Research 2

1.3 Aim of the Study 3

1.4 Objectives of the Study 3

CHAPTER TWO 4

2.0 LITERATURE REVIEW 4

2.1 Taste and Odor of Water 6

2.2 Potable Water 6

2.3 Sources of Water 7

CHAPTER THREE 9

3.0 METHODOLOGY 9

8
3.1 Sample Collection 9

3.2 Sterilization of Materials 10

3.3 Preparation of Media 10

3.3.1 Nutrient Agar 10

3.3.2 Lactose Broth 10

3.3.3 Eosin Methylene Bine (EMB) Agar 11

3.4 Analysis of Samples 11

3.4.1 Physicochemical Analysis 11

3.4.2 Bacteriological Analysis 13

3.4.2.1 Presumptive Test 13

3.4.2.2 Confirmatory Test 13

3.3.1.1 Pure Culture 14

3.5 Biochemical Tests 14

3.5.1 Gram’s Staining 14

3.5.2 Catalase Test 15

3.5.3 Motility Test 15

3.5.4 Oxidase Test 16

3.5.5 Sugar Fermentation Test 16

3.5.6 Citrate Utilization Test 16

3.5.7 Indole Test 17

CHAPTER FOUR 18

6
4.0 RESULTS 18

CHAPTER FIVE 25

5.0 DISCUSSION, CNCLUSION AND RECOMMENDATION 25

5.1 Discussion 26

5.2 Conclusion 26

5.2 Recommendations 26

REFERENCES 27

8
 LIST OF TABLES

Table 1: Atmospheric condition during storage period 19

Table 2: The pH value of water sample during storage 20

Table 3: Temperature of the Water Samples During Storage 21

Table 4: Turbidity of the Water Samples 22

Table 5: Colour of the Water Samples 23

8
 ABSTRACT

In this study, different colour of containers were used to stored water for eight days in
different environments in which some were stored indoor and some outside where there
was no shield. Sterilized materials were used in the preparation of media such as
Nutrient Agar, Lactose broth and Eosin Methylene Blue (EMB). Analysis of samples
involved Physicochemical analysis and Bacteriological analysis. Bacteriological
analysis involved Presumptive Test, Confirmatory Test and Pure culture. Biochemical
tests involved Gram’s staining, Catalase test, Motility test, Oxidase test, Sugar
fermentation test, Citrate utilization test and Indole test. It was find out that the water
sample after carrying out bacteriological analysis was found to have seven (7) different
species of microorganisms (Enterobacter sp., klebsiella sp, Citrobacter sp, micrococcus
sp, proteus sp, bacillus sp and seratia sp.). The temperature condition during the period
of storage varied between 29°C - 34°C and the temperature of the water sample during
storage varied between 24.0°C - 28.3°C which favored the growth of microbes but
differed with the findings of Eniola et al., (2006). There was no significant difference in
effect of the different color of containers and that the outdoor storage of water in light
colored container was found to be desirable because of its permeability that permits the
free flow of light through the container. It was concluded that the color of the container
and fluctuation in light and radiation are also important. From the result
recommendation were made that boreholes should be well located in a well sanitized
environment and away from soak ways to avoid fecal contamination of the borehole
water and that the environment were the borehole is located should constantly be
sanitized on a periodic bases

8
 CHAPTER ONE

1.0 INTRODUCTION

Nigeria, Africa’s most populated country, with about 150 million people,

has limited potable water supply (Punch newspaper, 2014). In the last 50 years or

so, Nigeria’s population has steadily been on the increase, a development which

has become a huge concern to economic planners in the country. Nigeria’s large

population has brought about undue pressure on the available infrastructure across

the country, many of which have been stretched to the very limit. The Nigerian

Minister of Water Resources was reported to have revealed that about 70 million

Nigerians lacked access to potable water. (Punch newspaper, 2014). The supply of

water in terms of quality, quantity, when and where it is needed continues to

generate concerns Eniola, et al, (2006). Water plays a significant role in the

socioeconomic development of human populations (Michael, 1998). Wells,

boreholes and springs that are properly located produce water of very good quality

(Gerald et al., 1992). However, it must not be taken for granted that ground water

will always meet the WHO standards for drinking water (Maggy, et al., 2005).

Rogbesan, et al, (2002) reported heavy bacteria load in water from some

boreholes in Ilorin.

The random supply of water has made water storage a common practice

among individuals and households, especially when there is pressure on the water-

source. (Maggy, et al., 2005 and Eniola, et al., 2006) highlighted the importance

of a few days of indoor storage in improving the physical and bacteriological

10
quality of borehole water. In many communities in Nigeria, in which Owo is one,

it is common to pump water into overhead storage tanks. The most common are

the plastic tanks of different colors, which are usually placed outdoors.

The outdoor location of the water tanks exposes them to solar radiation. In

addition to generating heat, many forms of radiations are harmful to

microorganisms; low levels of ionizing radiations will produce mutations and may

indirectly result to death, whereas higher levels are directly lethal (Penner, et al.,

2002). Even visible light when present in sufficient intensity can damage or kill

microbial cells (Prescott, et al., 1999). The amount of radiation available is

affected by aerosol optical depth and cloud parameters (Sekinguchi, et al 2003):

Climatic condition is affected by scattering of short waves, adsorption of solar and

long wave radiations (King et al., 1999; Penner et al., 2002; Takemura et al.,

2002).

1.1 Research Problem

The lack of potable water supply has made the practice of water storage a

habit amongst Nigerian homes without proper treatment of the water which

exposes the consumers to pathogenic microorganisms.

1.2 Justification of Research

This research establishes a proof on the effect of the color of container and

storage condition on bacteriological quality of borehole water. The outcome of

8
this research would be of great importance to man both directly and indirectly in

the sense that the result of this research would determine the importance of long

time storage of water.

1.3 Aim of the Study

The study is aimed at investigating the effect of the color of container and

storage condition (indoor and outdoor) on the bacteriological and

physicochemical quality of borehole water.

1.4 Objectives of the Study

The objectives are as follows:

 To determine the effect of outdoor storage and indoor storage of borehole

water on the bacteriological quality of borehole water.

 To compare the physicochemical result of both the water stored indoor and

those stored outdoor.

 To compare the effect of the color of container stored indoor and those

stored outdoor.

12
 CHAPTER TWO

2.0 LITERATURE REVIEW

Water is a transparent fluid which forms the world’s streams, lakes, oceans

and rain, and is the major constituent of the fluids of living things (Prescott et al.,

2019). As a chemical compound, a water molecule contains one oxygen and two

hydrogen atoms that are connected by covalent bonds. Water is a liquid at

standard ambient temperature and pressure, but it also co-exists on Earth with its

solid state, ice; and gaseous state, steam (water vapor) (John, 2000).

Water covers 71% of the earth surface, it is vital for all known forms of

life. On Earth, 96.5%.of the planet’s water is found in seas and oceans, 1.7% in

groundwater, 1.7% in glaciers and the ice caps of Antarctica and Greenland, a

small fraction in other large water bodies, and 0.001% in the air as vapor, clouds

(formed of solid and liquid water particles suspended in air), and precipitation.

Only 2.5% of the Earth’s water is fresh water, and 98.8% of that water is in ice

and groundwater. Less than 0.3% of all freshwater is in rivers, lakes, and the

atmosphere, and an even smaller amount of the Earth’s freshwater (0.003%) is

contained within biological bodies and manufactured products (Gerald, et al.,

2012).

Water on earth moves continuously through the water cycle of evaporation

and transpiration (evapotranspiration), condensation, precipitation and runoff

usually reaching the sea. Evaporation and transpiration contribute to the

8
precipitation over land. Water used in the production of a good or service is

known as virtual water (John, 2000).

Safe drinking water is essential to humans and other life forms even though

it provides no calories or organic nutrients. Access to safe drinking water has

improved over the last decades in almost every part of the world, but

approximately one billion people still lack access to adequate sanitation. There is

a clear correlation between access to safe water and gross domestic product per

capita. However, some observers have estimated that by 2025 more than half of

the world population will be facing water-based vulnerability. A report issued in

November 2009, suggests that by 2030, in some developing regions of the world,

water demand will exceed supply by 50%. Water plays an important role in the

world economy, as it functions as a solvent for a wide variety of chemical

substances and facilitates industrial cooling and transportation: Approximately

70% of the fresh water used by humans goes to agriculture (John, 2000).

The major chemical and physical properties of water are:

 Water is a liquid at standard temperature and pressure. It is tasteless and

odorless. The intrinsic color of water and ice is a very slight blue hue,

although both appear colorless in small quantities. Water vapor is

essentially invisible as a gas (Prescott, et al., 1999).

 Water is transparent in the visible electromagnetic spectrum. Thus aquatic

plants can live in water because sunlight can reach them infrared light is
14
 strongly absorbed by the hydrogen-oxygen or OH bonds (Prescott, et al.,

1999).

 Pure water has a low electrical conductivity, but. this increases with the

dissolution of a small amount of ionic material such as sodium chloride

(Prescott et al., 1999).

2.1 Taste and Odor of Water

Pure water is tasteless and odorless. Water can dissolve many different

substances, giving it varying tastes and odors. Humans, and other animals, have

developed senses that enable them to evaluate the potability of water by avoiding

water that is too salty or putrid.

The taste of spring water and mineral water, often advertised in marketing

of consumer products, derives from the minerals dissolved in it. The advertised

purity of spring and mineral water refers to absence of toxins, pollutants, and

microbes, not to the absence of naturally occurring minerals (Prescott et al.,

1999).

2.2 Potable Water

This is referred .to a type of water which is fit for consumption by humans

and other animals. It is also called drinking water, in reference to its intended use.

Water may be naturally potable, as is the case with pristine springs, or it may need

to be treated in order to be safe. In either instance, the safety of water is assessed

8
with tests which look for potentially harmful contaminants (WHO, 1996).

The issue of access to potable water is very important. In developed

countries, people may not put a great deal of thought into the source of their

water. In many First World nations, citizens can turn on a tap for fresh, potable

water which may also be enriched with substances for health. In developing

countries, however, and especially in Africa, a large proportion of the population

does not have access to safe water.

Water which is not safe to drink can carry diseases and heavy metals.

People who consume this water will become ill, and there is a risk of death.

Unfortunately, even in areas where the water is known to be unsafe, people may

drink it anyway^ out of desperation (Olayemi, et al., 2005).

2.3 Sources of Water

Nigeria presently has a population in excess of 150 million. However,

some reports suggest that less than a fifth of this population has access to clean

pipe borne drinking water. The issue of provision of good quality drinking water

has been the source of several efforts by the government and nongovernmental

organizations. This is to alleviate the suffering due to poor drinking water quality.

Drinking untreated water can result in several unwanted effects for the

individuals, communities and ultimately for the nation (Prescott, et al., 1999).

Typically, Nigerians resort to streams, wells or bore-holes as a source of

16
water. Although ground water (depending on the depth) can be a source of “clean”

water, this raw water should still be tested and ultimately treated prior to usage.

The streams, borehole and wells are easily exposed to heavy minerals which on its

own are carcinogenic like arsenic, cadmium, cobalt, lead etc. due to the presence

of this pollutants, it is necessary to test and perhaps treat this water prior to use.

However this is presently not the case, due to lack of access to water test kits as

well as treatment options (Rogbesan, et al., 2002).

Ground water can also be contaminated by biological agents depending on

the source. Shallow boreholes and wells can be exposed to contamination from

sewage systems, thereby increasing the risk of presence of water-borne diseases.

In Nigeria, due to the poor sewage systems, this risk is significantly enhanced.

These water-borne agents can be detected by proper analytical tests, followed by

water treatment (Salle, 2013).

8
 CHAPTER THREE

3.0 METHODOLOGY

3.1 Sample Collection

The borehole water sample used for this project was collected into tap

fitted plastic containers as described by WHO (2000). Blue, transparent, black,

and green colored containers with a duplicate each were used to store the water

sample and the containers were designated as B1 (Blue container outdoor), T1

(transparent container outdoor), BLK1 (black container outdoor) and G1 (green

container outdoor) while their duplicates as B2 (blue container indoor), T2

(transparent container indoor), BLK2 (black container indoor) and G2 (green

container indoor). The Bl, Tl, BLK1 and G1 borehole water sample were stored

outdoor without shade covering while the B2, T2, BLK2 and G2 were stored

indoor for a total period of eight (8) days. Water sample was also collected in a

sterile bottle labeled as Control Water Sample (CWS). All necessary analysis

(Physicochemical analysis, bacteriological analysis, and biochemical test) were

carried out on this fresh water sample and results were recorded in order to be

compared to the results of the analysis that would be carried out on the colored

containers stored outdoor and indoor at intervals of four (4) days for a total

storage period of eight (8) days!

18
3.2 Sterilization of Materials

All glass wares such as borosilicate Petri dish, beakers, conical flasks etc

were thoroughly washed with detergents, rinsed with potable water and allowed to

dry. After drying, the glass wares were placed in hot air oven to attain sterility at

1800C for three (3) hours. Inoculation table were well sterilized using ethanol and

cotton wool and a spirit lamp placed on the center of the inoculation table before

the working material and water sample was brought unto the table.

3.3 Preparation of Media

3.3.2 Nutrient Agar

This media was used for the enumeration and isolation of bacteria. The

concentration of each medium is dependent on the manufacturer’s directive. 28g

of powdered nutrient agar was weighed and dissolved into 1000ml of distilled

water and stilted vigorously to attain an even mixture using magnetic stirrer, the

medium was then autoclaved at 1210C for 15 minutes in order to attain sterility.

3.3.2 Lactose Broth

The medium was produced with double strength capacity for easy detection

of the slightest presence of microbes. 13g of lactose broth powder was weighed on

a weighing balance and dissolved into 1000ml of distilled water, it was then

stirred using a magnetic stirrer in order to achieve an even mixture. 10ml was then

measured each into 40 McCartney bottles containing an inverted Durham’s tube,

8
sealed and then autoclaved at 1210C for 15minutes to attain sterility.

The manufacturer’s directive was followed and multiplied by two in order

to attain a double strength medium

3.3.3 Eosin Methylene Bine (EMB) Agar

This is a differential and a selective media used for the growth of coliform

microorganism. Following the manufacturers directive, 37.5g was weighed and

dispensed into 1000ml of distilled water and then stirred using a metal stirrer.

After attaining an even mixture, it was autoclaved at 1210C for 15minutes,

allowed to cool off a little and then dispensed into the Petri dishes. The Petri

dishes were rocked gently to attain an even distribution of the agar in the plates

and then allowed to solidify.

3.4 Analysis of Samples

3.4.1 Physicochemical Analysis

The control water sample (CWS) was the first sample to be analyzed.

Physical analysis was first carried out. A little quantity of the sample (CWS) was

poured into the beaker and the temperature was observed using conductivity

meter, the result was recorded. pH meter was calibrated and put inside the water

sample (CWS) after a minute; the result displayed on the pH meter was also

recorded. The turbidity of the water sample was observed and result recorded

using turbidity meter. The color of the water checked for, using
20
spectrophotometer (DR 5000), and the result was also recorded. Dissolved oxygen

(D02I of the water sample was recorded using D02 meter. The parameters looked

out for in the chemical analysis carried out was total hardness and total alkalinity.

Total hardness test was carried out to determine the capacity of water to react with

soap. Hard water requires considerably more soap to produce lather. Hard water

often produces a noticeable deposit of precipitate (e.g. insoluble metals, soaps or

salts) in containers, including “bathtub ring”. It is not caused by a single substance

but by a variety of dissolved polyvalent metallic ions, predominantly calcium and

magnesium other cations (e.g. Aluminum, barium, iron, manganese, strontium and

zinc) also contribute. Hardness is most commonly expressed as milligrams of

calcium carbonate equivalent per liter. Water containing calcium carbonate at

concentrations below 60 mg/1 is generally considered as soft; 60-120 mg/1,

moderately hard; 120-180 mg/1, hard; and more than 180 mg/1, very hard

(McGowan, 2000). Although hardness is caused by cations, it may also be

discussed in terms of carbonate (temporary) and non-carbonate (permanent)

hardness.

100ml of the water sample (CWS) was measured and dispensed into a

conical flask using a measuring cylinder then 20 drops of k-10 buffer was

introduced into the l00ml of water sample, this is to stabilize the pH, 5 drops of

net solution, was then added which produced a lilac color. EDTA was poured into

the burette and titrated against the water sample until color change was observed

i.e. the water sample changes from lilac color to light blue as the end point. The

reading of the titration was then recorded.

8
3.4.2 Bacteriological Analysis

10ml of the water sample was measured using a sterile syringe and

introduced into five (5) already autoclaved McCartney bottles containing inverted

Durham’s tube and 10ml each of double strength lactose broth. The bottles were

well labeled to detect the slightest presence of gas bubble in the Durham’s tube.

0.1ml and 1ml of the water sample was measured into two batches of McCartney

bottles (each batch contains five bottles) containing autoclaved single strength

lactose broth and inverted Durham’s tube. The bottles were incubated at 37oC for

24 hours. After the incubation period, positive bottles exhibited the presence of

gas bubble inside the Durham’s tube, color change and turbid nature of the

samples while negative bottles has no gas bubble and wasn’t turbid.

3.4.2.1 Presumptive Test

After the incubation period, the positive McCartney bottles were selected

and cultured on nutrient agar (using pour plate method) in order to get the total

bacteria count using Most Probable Number (MPN). The same positive bottles

were streaked on EMB (Eosin methylene blue) in order to test tor the presence of

E. coli in the water sample. The EMB agar was incubated for 24 hours at 450C.

3.4.2.2 Confirmatory Test

The Positive plates from presumptive test were sub-cultured onto Nutrient

Agar using streak method and incubated at 370c for 24 hours. Colonies growth

22
was observed 24 hours of incubation.

3.3.2.1 Pure Culture

The pure isolates sub-cultured from Nutrient Agar plate were inoculated on

slant nutrient agar bottles for further analysis (biochemical tests) and preservation.

3.5 Biochemical Tests

3.5.1 Gram’s Staining

Christain Gram’s technique was the method used to carry out Gram’s

staining. A clean grease-free slide was used and a smear was made on the slide

using a wire loop that has been flamed to red hot and whipped to cool to drop

normal saline on the slide. The wire loop was flamed again to red, whipped to cool

off and used to pick the isolate. The isolate was emulsified with the normal saline

on the slide and then allowed to air dry, it was then slightly passed through flame in

order to fix the smear on the slide. 1 he fixed smear on the slide was then flooded

with crystal violet, allowed to stain for 30-60 seconds and gently washed off under

running water, drained and then flooded with 95% ethanol, washed off immediately

with water and drained. The ethanol decolorizes the smear. Finally, Safranin which

is referred to as a counter-stain was added and allowed to stain for 2 minutes,

washed off with water and then drained. The back of the slide was wiped with

cotton wool and allowed to dry to permit easy viewing under the microscope with

XI00 objective lens (oil immersion). (Microbiologybytes.com, 2014).

8
3.5.2 Catalase Test

This test is used to identify organisms that possess the enzyme catalase

which breaks hydrogen peroxide into water and oxygen. (Amrita, 2014).

2H202 2H20 + 02

A loopful of a 24 hour-old bacteria culture was smeared on a slide and a

drop of 3% hydrogen peroxide was added to the mixture and then examined for

the formation of air bubbles.

3.5.3 Motility Test

Semi-solid medium was prepared using half strength nutrient agar (14.0g -

1litre of distilled water). The medium was stirred using magnetic stirrer to attain a

homogenized mixture. 10ml each was dispensed into clean test tubes corked and

sterilized at 12IOC for 15 minutes at lkg/cm3 pressure. A 48 hour culture of the

unknown bacteria was then inoculated by stabbing the isolates separately in the

medium using a sterile inoculating needle and incubated at room temperature for

48 hours. A test tube not inoculated but was also kept at room temperature for 48

hours to serve as control.

Motility was observed as growth outside the line of inoculation in the

medium (Olutiola, et al., 2000).

24
3.5.4 Oxidase Test

Two drops of 1% freshly prepared oxidase reagent was placed on a filter

paper in a clean Petri dish. The test organism was smeared on it with a glass rod.

A blue - purple color development within 10 seconds was recorded as a positive

result and in the absence of blue - purple color a negative result was recorded.

3.5.5 Sugar Fermentation Test

The bacteria isolates was tested for their fermentation of various sugar, the

medium used consisted of peptone 3.0g, 1 %(1.0g) solution of the desired sugar

(lactose, mannitol, glucose and fructose) and 2ml of 0.01% phenol red (indicator)

all dissolve in 100ml of distilled water. 10ml of the solution was dispensed into

test tubes. A Durham’s tube was inversely inserted into each of the medium and

sterilized at 1150C for 10 minutes. Each of the test tubes was inoculated with die

bacterial isolates. An uninoculated tube serves as control. Incubation was carried

out at 370C for 24 hours. Acid production was shown by a change in color of the

medium to red or yellow (positive), while gas production was shown by a

displacement of the solution in the Durham’s tube by air.

3.5.6 Citrate Utilization Test

Citrate agar slant was prepared in a McCartney bottles, the surface of the

slope was streaked using a sterile wire loop, and the butt of the agar was stabbed

with a saline suspension of the test organism and incubated at 35oC for

8
48hours.positive citrate shows bright blue color change while the negative citrate

shows no color change. (Microbeonline.com, 2014).

3.5.7 Indole Test

The bacteria culture was inoculated in peptone water and incubated at 35-

37oC for up to 48hours. 0.5ml of Kovacs reagent was added shake gently and was

examine for the appearance of red color in the surface layer within l0 minutes

which shows indole positive while negative test retain color of broth. (Microbe

Library, 2009).

26
 CHAPTER FOUR

4.0 RESULTS

A total of seven (7) bacterial species (Enterobacter sp, Klebsiellci sp,

Citrobacter sp, Micrococcus sp, Proteus sp, Bacillus sp and Seratia sp) were

identified after the first analysis (prior to storage), the bacteriological analysis

carried out on the water samples after four days of storage showed a decrease in

the total number of microbes present in each of the storage containers and after

eight (8) days of storage the microbial load became less than the microbial load in

the previous storage intervals (prior to storage and four days after storage).

Table 1 is the result for atmospheric condition (temperature, sky condition

and relative humidity) of the storage period. The relative humidity ranged from

81.08 - 83.04%, the sky condition was generally cloudy which supported an even

distribution of sunlight and the temperature ranged between 290C >- 340C All the

results gotten on this table support the proliferation of microbes in water

8
Table 1: Atmospheric condition during storage period

Storage period (days) Atmospheric condition

Relative humidity (%) Sky condition temperature

1 83.04 Drizzling 29

4 81.0 Cloudy 32

8 81.84 Cloudy 34

Table 2 is the pH value of the water sample during the period of storage.

The water sample maintained a steady pH (6.5) prior to storage. The pH value

during storage period ranged from 6.5-7.0 which falls within the range that would

favor bacteria proliferation (Atlas, 1995). The observed increase in pH during

storage could be due to the activities of resident flora or their death, which results

in the release of inorganic substances such as ammonia (Rogbesan et al., 2002).

WHO standard for potable water pH is 6.5-8.5.

28
Table 2: The pH value of water sample during storage

Storage periods (days) pH

Blue Transparent Black Green

container container container container

B1 B2 T1 T2 BLK1 BLK2 G1 G2

0 6.5 6.5 6.5 6.5 6.5 6.5 6.5 6.5

4 6.6 6.8 6.5 6.9 6.6 6.7 6.5 6.6

8 6.7 6.9 6.7 7.0 6.7 6.8 6.6 6.9

B1 = Blue container stored indoor. B2 = Blue container stored outdoor

T1=Transparent container stored indoor. T2 = Transparent container stored

outdoor

BLK1=Black container stored indoor. BLK2 = Black container stored outdoor

G1=Green container stored indoor. G2 = Green container stored outdoor

Table 3 shows the varying temperature of the water samples during

storage. The temperature ranged from 24.00C - 28.30C which supports bacterial

proliferation. The rise and fall of the temperature readings could be due to the

general atmospheric temperature.

8
Table 3: Temperature of the Water Samples During Storage

Storage periods Temperature (0c)

(days)
Blue Transparent Black Green
container container container container

B1 B2 T1 T2 BLK1 BLK2 G1 G2

0 27.1 27.1 27.1 27.1 27.1 27.1 27.1 27.1

4 27.0 27.8 27.3 28.0 26.9 28.3 27.2 28.0

8 26.8 24.3 26.5 24.0 26.2 24.7 26.2 24.6

Bl=Blue container stored indoor. B2 = Blue container stored outdoor

Tl = Transparent container stored indoor. T2 = Transparent container stored outside

BLK1 = Black container stored indoor. BLK2 = Black container stored outdoor

G1 = Green container stored indoor. G2 = Green container stored outside

Table 4 shows the turbidity test result of the water samples during the

period of storage. It would be noticed that the turbidity result ranged from 1.00 -

1.09 which is within the WHO acceptable range (< 5). The progressive reduction

in the turbidity test result is similar to the observations of Olayemi et al., (2005)

and Eniola et al., (2006).this has been attributed to gravitational pull, which

causes suspended materials to settle out of the water over time.

30
Table 4: Turbidity of the Water Samples

Storage periods (days) Turbidity

Blue Transparent Black Green

container container container container

B1 B2 T1 T2 BLK1 BLK2 G1 G2

0 1.09 1.09 1.09 1.09 1.09 1.09 1.09 1.09

4 1.07 1.05 1.06 1.02 1.08 1.06 1.07 1.04

8 1.00 1.01 1.00 1.00 1.02 1.01 1.01 1.02

Bl=Blue container stored indoor. B2 = Blue container stored outdoor

Tl = Transparent container stored indoor. T2 = Transparent container stored outside

BLK1 = Black container stored indoor. BLK2 = Black container stored outdoor

G1 = Green container stored indoor. G2 = Green container stored outside

The WHO standard for potable water color is <15. Wells, boreholes and

springs that are properly located produce water of very good quality (Gerald et al.,

2012). The below range (b/r) recorded in the test for color supports the fact that

the borehole is located in a well sanitized environment.

31
Table 5: Colour of the Water Samples

Storage periods (days) Colour

Blue Transparent Black Green

container container container container

B1 B2 T1 T2 BLK1 BLK2 G1 G2

0 b/r b/r b/r b/r b/r b/r b/r b/r

4 b/r b/r b/r b/r b/r b/r b/r b/r

8 b/r b/r b/r b/r b/r b/r b/r b/r

b/r = below range. The WHO accepted range for water color is <15

B1 = Blue container stored indoor. B2 = Blue container stored outdoor

T1 - Transparent container stored indoor. T2 = Transparent container stored outdoor

BLKl = Black container stored indoor. BLK2 = Black container stored outdoor

G1 = Green container stored indoor. G2 = Green container stored outdoor

The biochemical characteristics of microorganisms present in water was

determined using biochemical tests. Bergey’s manual of determinative

bacteriology was used to easily determine the suspected organism.

32
 Control water sample

Suspected Gra Ga Oxidas Citrat Indol Motilit Glucos Lactos Sucros

organism m s e test e test e test y e e e
rxn test

Enterobacte + + + + - + + + +
r sp

Klebsiella - + - + - - + + +
sp

Citrobacter - + - + - + + + +
sp

Micrococcu + - + + - - - - -
s sp

Proteus sp - + - + + + + - +

Bacillus sp + - + + - + + - +

Seratia sp - + - + - + + - +

+ = Positive result

- = Negative result

An equal number (7) of bacteria was recorded in the water sample prior to

Storage. The population of bacteria declined with length of storage. Containers

stored outdoor had less bacteria count than those stored indoor and the transparent

container had the lowest bacterial count. Citrobacter and proteu ssp survived the

indoor and outdoor storage after the total storage period (8 days).

33
 CHAPTER FIVE

5.0 DISCUSSION, CNCLUSION AND RECOMMENDATION

5.1 DISCUSSION

The water sample after carrying out bacteriological analysis was found to

have seven (7) different species of microorganisms (Enterobacter sp, Klebsiella

sp., Citrobacter sp, micrococcus sp, Proteus sp, Bacillus sp and seratia sp). The

temperature condition during the period of storage varied between 290C — 340C

and the temperature of the water sample during storage varied between 24.00C -

28.30C which favored the growth of microbes but differed with the findings of

Eniola et al., (2006). The pH fell within the range that would favor bacterial

proliferation which is similar to the observation by Atlas (2015). The observed

increase in pH during storage could be due to the activities of the resident flora

and or their death, which results in the release of inorganic substances such as

ammonia (Rogbesan et al., 2002).

The zero total coliform count shows the borehole is free of fecal

contamination which coincides with Gerald et al., (2012), that says boreholes and

springs that are properly located produce water of very good quality. The

reduction in population of total bacteria as the day of storage increased is similar

to the observation by Payment et al., (2015). Decline in the bacterial population

can be attributed to death of the resident bacteria during the storage period due to

depletion of nutrients which was stated and also observed in the work of Olayemi

et al., (2005). Reduction in the bacterial load was more prominent in buckets

34
stored outdoor, this is attributable to direct radiation from sunlight on the buckets

outside. Among those outdoor, penetration by radiations would be more

pronounced in the transparent bucket which results in the death of the microbes

present in the water sample and this coincides with the work of Eniola et al.,

(2006).

5.2 Conclusion

This study buttress that storage is valuable as a preliminary accessory stage

of treatment but it cannot be relied on as a sole measure of purification. The color

of the container and fluctuation in light and radiation are also important. There

was no significant difference in effect of the different color of containers. This is

attributable to cloud cover that creates a uniform temperature condition. In

conclusion, outdoor storage of water in light colored container was found to be

desirable because of its permeability that permits the free flow of light through the

container.

35
5.3 Recommendations

With regards to the results achieved from the whole analysis, i recommend

the following:

 Boreholes should be well located in a well sanitized environment and away

from soak way’s to avoid fecal contamination of the borehole water.

 Boreholes should be dug to a minimum depth of 120m to obtain potable

water

 Borehole water should be periodically taken to the laboratory for E

analysis to check for its potability standard

 It is recommended that borehole water should be stored outdoors in light

colored containers for a period of 8 - 10 days as its storage I would

contribute to the reduction of bacterial load.

 It is also recommended that the environment were the borehole is located

should constantly be sanitized on a periodic bases.

36
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