CHAPTER ONE

1.0 Introduction

Bacteriology as an aspect of studying bacterial becomes a vital and

significance in human populations as a question in our sources of water,

perhaps the needs to identify those bacteria remains an important concept today

in our environment. Many infectious diseases are transmitted by water through

the fecal-oral route. Disease contacted through drinking water kills about five

(5) million children annually and make 1/6th of the world population sick

(WHO, 2004). Water is the basics of all life and primary needs for vital

processes with increasing urbanization, industrialization and population growth,

and water sources available for various purposes such as drinking, recreation,

agriculture and aquaculture have been adulterated with industrial as well as

animals and domestic waste. As a result it has become the most important

means of transmission of several infectious disease. In Nigeria majority of the

rural populace do not have access to portable water and therefore depend on

wells, for domestic used. The bacterial qualities of ground water and other

natural water supplies in Nigeria have been reported to be unsatisfactory, with

coliform count far exceeding the level recommended by WHO (Dada et al,

1999a, 1999b Edema et’ al, 2001). About one third draws water from

unprotected wells, Drinking water supply is liable to contamination with

1
sewage and other excreted matter, which may cause outbreak of intestinal

infections such as typhoid fever, choler, and other diarrheal diseases (Feachem,

2009). Several researchers (Ekundayo, 1977, Fleisher et’ al 1998, Okafo et’ al

2003) have documented the health significance of fecal pollution of water

bodies, the indiscriminate dumping of untreated wastes in our water bodies

brings about physical, chemical and biological deteriorations of such water

bodies especially when these discharges are beyond their natural purifying

capacities (Horvat et’ al and Fujioka et al, 2011).

1.1 Aim of the Study

The aim of the study is to carry out microbiological investigation of well water

of different location in Potiskum local government Area.

2
 CHAPTER TWO

2.0 Literature Review

The study of well water characteristics for different purpose has become

very essential, because of its interplay between livers. But a multiplicity of well

water characteristics is encountered in nature, this is more significant from a

chemical, point of view than from physical perspective from relatively clear and

pure rain water with little dissolve impurities to highly contaminated surface

water characteristic, this is because percolation through table (Ademorotti

1996). Research has shown that drinking water quality represent the

concentration of a constitute that does not result in any significant risk to

contaminated over a life time of consumption of well water. If part of the faeces

get in to water it can cause epidemic that can read to loss of lives (Ademorratti

1996).

2.1 Drinking Water and Infectious Diseases

Water is essential element for living. Ironically, human with their

activities keep degrading its quality and quantity. Unsafe water, inadequate

quantities of water, poor sanitation and hygiene are leading to infectious

diseases and children are the most vulnerable group to be infected. Globally,

every year there are more than 1.5million children within the age of five and

3
below are died mostly because of diarrhea (Unicep, 2006). Infectious disease

isone that can be transmitted from one to anther and caused by microorganisms

such as bacteria and viruses. These microorganisms are using water and air as

their medium transportation to be transmitted form host to the new victims

(Caincross and Feachemi 1993: Westrell, 2004) it is only in time of water

shortage that man begins to realize how totally depend all living things are on

water. Because it is the only common liquid on our plannet we are almost over

familier with it, and tends to forget many of the remarkable properties on most

of which our very existence depend (Baron 2007).

Sale, (1967) started that, most water contains larger number of bacteria,

but the number may vary considerably depending up on the source of the water

e.g deep or shallow walls, spring, rivers, lakes, ponds, or stream water polluted

with sewage may contain thousands or even millions of organisms per

millimeter. Under some conditions, disease organisms may sometimes be

present in water. Naturally fewer bacteria occur in sea water than in soil, which

is due its having poor quality of being a culture medium. Water that is

contaminated serves as a mechanism of transmitting communicable diseases

such as the typhoid, cholera, dysentery, diarrhea disease and guinea worm

infection. This reads millions of people to get expose to dangerous level of

biological contaminants and chemical pollutants in their drinking waters, which

4
infect due to inadequate management of the urban industrial or Agricultural

waste water (WHO 2012). According to the WHO report (2012), an estimated

total cause of diarrhea disease that claimed live in 2008 was 2.5 million people.

But for children under five, the burden was greater than the combine burden of

HIV/AIDs and malaria. A total of 58 countries from all continents reported a

cumulative total of 589, 854 cholera case in 2011, representing an increase of

85% from the 2010 record. The greatest proportion of cases was reported form

the island of Hispaniola and the African continent. These trends reflects the

need to shift from basic responsiveness to a comprehensive, multidisplinary

approach that works with communities to improve access to safe drinking water

and sanitation, evourage sporing anaerobes (Clostridium perfringens) and

members of coliform group (Okoro, 2002).

According to Okoro (2002), the corfom bacilli are considered as the most

variable indications of faecal pollution because they are likely occur in small

numbers even in water supplies far removed from the possibility of human

contamination.

2.1 Source of Pollution of Water in Nigeria

In Nigeria, majority of the rural populace do not have access to potable

water and therefore depend on well, stream and river water for domestic use.

5
The bacterial qualities of groundwater, pipe borne water and other nature water

supplies in Nigeria, have been reported to be unsatisfactory, with coliform

counts far exceeding the level recommendation by WHO (Dada et’ al 1999a,

199b, Edema et’ al 2001). A number of studies (World Bank 1993 Brockchoff

1995, Hoddinott 1997) have at one time or the other examines impact of water

pollution on the variables that determine the health status of the household

numbers. Most of the studies hypothesize that an improvement of water quality

has direct effects on people’s health via reduce exposure to water associated

disease. Hospital record has confirms high incidence of water borne diseases in

the urban settlement of Nigeria of all the cost of environmental degradation,

damage to human health is by far highest. There is direct link between urban

environmental degradation and public health in terms of water related disease

such as diarrhea, dysentery, cholera and typhoid fever. The rapid growths of

unstructured infrastructures and social services have created an environmental

situation in many part of the country which becoming inimical to healthy living

behavioral change and promotes the targeted use of oral cholera vaccines where

the diseases become endwmic. As though consumption of contaminated water

represents the greatest risk, other roles of transmitted can lead to disease and

contribute to the disease burden, WHO estimated that more than 2000 million

more are affected by schistosomiasis and around 800 million more are at risk of

6
infection (WHO 2011). In 2006, the WHO reported that, in 2005 half of the

populations have been living in water stressed areas. This was because was

increasingly, countries have been utilizing waste water as an important source

of irrigation water and nutrients for crop production, the practice of which poses

health risk from exposure to untreated and partially treated waste for workers,

communities and consumers that need to be weighed against the potential

benefits of increased food production in term of improved nutrition and income.

However pathogens effects are caused by micro-organisms, where

bacteria, viruses and fungi are present in suffucuent numbers to cause health

hazard (Dix, 1994)- Bacterial contaminant from one of the indicator of

biological pollution high faecal coliform coult is mainly due to domestic waste

(chua et al-1997) as well as animal wastes pollution of water (choll, 1994 Law

et al 2001a). according to Roberts (2006) and Collins (2000), bacteriological

water analysis is a method of analyzing water to estimate the numbers of

bacteria present and, it needed to findout what sort of bacteria they are it is

seldom possible to isolate enteric pathogens directly from contaminated water

because they are usually present on small numbers, unless contaminated from

an infected individual has been recent and massive. The bacteria that have

principally served as indices of such contamination are faecal streptococci

(Streptococcus Fecalis).

7
2.2 Source of Water Contamination in Potiskum

Potiskum like any other Local Government in Yobe State is facing

serious challenges over provision of safe and adequate water supply. Increase in

population due to mainly rural-urban migration, insecurity and population

growth meant on increase in the use of water in the two surface and

groundwater contamination, which is due to influx of agriculture fertilizer

runoff that contains nitrates, and improperly isolated landfills may introduce a

variety of organic substances many of which are bridgeable and any lead to

bacterial pollution in ground water to become excessive. Water sources are

subject to variety of bacterial contaminates introduced by storm water runoff

over the water shed and the upstream discharge of domestic and industrial

waste (Edema et’ al 2001).

2.3 Portability and Quality of Drinking Water

The most common and widespread danger associated with portable water

is contamination, either directly or indirectly by sewage, or by human and

animals activities (WHO1984) water portability depends on the treatment

passed on the water before drinking in order to remove contaminates. Some

8
water borne diseases “parasites that cause illness with the symptoms of

diarrhea, nausea and abdominal cramp are microscopic and they include

giardiasis (Boron 2010). Safe and portable water according to (Tebulle 1971)

should conform to the following qualities or characteristics.

a) Free from pathogenic organism

b) Be clear

c) Should not be salty

d) Free of compounds that have offensive taste or odour among other.

2.4 Pathogens Associated with Polluted Water

The use of waste water for domestic and irrigation purposes has been

responsible for many diseases outbreak caused by bacteria, protozoa, parasitic

helminthes and viruses (Bitton 19980, Parasher and Monroe 2001).

2.4.1. Esherichia Coli

Escherichia coli is a member of the genus enterobacteriaceae and most

stain are inhabitants of the intestinal tract and are practically always present in

faeces and thus also in faecally contaminated water. This has resulted in the

almost universal use of E. coli as a standard indicator for faecal contamination

(Franics et’ al 1999) the pathogenic strains commonly resulted in hemorrhagic

colitis gastroenteritis and kidney failure (Francis et’ at 1999).

9
2.4.2 Shigella

Chigella is another widespread water borne pathogen of the family

entrobacteriaceaes. The four species namely S. Sonnei, S dysenteries, S. boydi

S. flexneri are pathogenic. Shigella are regarded as rather fragile organisms

which do not servive well outside their natural habitats. However, some stains

are capable of survival below PH 6 and S. Sonnei can survive at temperature as

well as at low oxygen concentration (Gilland Selma 2006).

2.4.3 Salmonella

Salmonella is a pathogen of the family entrobacteriaceas. These bacteria

are cram-negative and rod shaped the genue comprises five pathosenic strains

namely S. typhimurium, S. enteriditis S. Saintpoul and S. minterideo (Francis

et’ al 1999).

Salmonella is a very resistant pathogen and it has a wide survival range.

It has adapted well to survive outside intestine particularly at water activities

between 0.43 and 0.52 (Maciorowski, 2007) Salmonellas grows optimally in

warm water temperature between 340C and 430C while its growth is

sulostantially retarded at 150C and generally prevented at 70C. this pathogen

alone is annually responsible for 1.3 million outbreaks of food and water borne

10
illness as well as for being the second biggest cause of diarrhea in the

developed and developing nations (Mena 2006).

2.4.4 Klebsiella and Proteus Speges

The gram – negative bacilli of genera Proteus and klebsiella are members

of the normal intestinal flora of human and warm blooded animals and may be

isolated from water and foods. With exception of Proteus, klebsiella is

sometimes referred to as the coliforms bacilli because of shared properties

particularly the ability to most species to ferment the sugar Lactose (Bernes,

2003).

11
 CHAPTER THREE

3.0 Materials and Methods

3.1 Study Area

The study was conducted at Potiskum Local Government about 96 kilometer

from Damaturu the capital city. It has an area of 551squarekilometer.

Geographically Potiskum is located at latitude 11,7119 (1142, 42, 84 N) and

longitude 11. 6697 (11410. 920E) and has an altitude of 415m. the town of

Potiskum has been a thriving trade hegemony in the state because of its

strategic Position as a center of commerce. Potiskum like all towns in the state

is facing the problem of lack of portable and adequate water supply that is safe

for human consumption, most of the inhabitants collect water in bucket from

hand pumps well while other from water vendors which are likely to be

contaminated

3.2 Sampling Stations

Water samples from eight wells from four different locations of varying

socio-economic and demographic status were randomly collected for

bacteriological examination. The areas are Nahuta (station A), Old Army

Barrack’s (station B) Unguwan Lamba (station C) and Kuyet (station D). Well
12
water constitutes the major source of drinking water in these areas. Most of the

wells under study areas were privately owned and are usually open to general

public half of the numbers of the studies wells were covered white the others

were not. Drawing of water from these wells was done by the use of 5 – 7 liter

containers tied directly to the well cover. In certain cases where this is not

possible. Individuals come with small bucket to draw water. The wells are not

Less than 10years old.

3.2 Laboratory procedures

3.2.1 Sterialization of the Glass Wares

The sterilization of glass wares such as sampling bottles, petridishes and test

tubes after washing with detergents was carried out in hot air oven and 160 0 for

2 hours (Harrigan, 1998).

3.2.2. Serial Dilution

The procedure of APHA (1985) and as adopted by Arzai (1995) was applied in

carrying out serial dilution.

3.2.3. Method of Serial Dilution

9mls of normal saline was poured into the test tubes and 1mc of the water

sample was transferred into the test tubes containing 9mls of the normal saline

13
which serve as a stock prepared as diluents. Serial dilution of 10-1 of the

dilluents was used from which subsequent of up to 10-4 was made (4-ford

dilution).

3.2.4. Sample Culturing for isolation of Bacteria Specie

The media used for this study was prepared by dissolving 28g in one liter

deionized water and Auto claving at 12101 for 15 minute.

3.2.5. Using Mc Conkey Agar

The Mc conkey agar this media was used to isolate the specie of

Escherichia coli and enteric coliforms by pour plate methods. The media was

prepared and the sample was put into the media and poured in the plate and

incubated at 3706 for 24 hours. After which the isolates were further

characterized by a combination of colonial and morphological characteristics on

solid media.

3.2.6 Using EMB Agar

The media was used to isolate the specie of salmonella and shigella

specie also was done by pour plate method. The media was prepared and the

sample was put into the media and poured in the plate and incubate at 370C for

24hours. After which the isolates were further characterized by a combination

of colonial and morphological characteristics on solid media.

14
3.2.7 Biochemical Characterization of Bacteria

Microorganisms especially bacteria were characterize by their ability to

produce various enzymes, the presence of various enzymes in turn can be

determined by the metabolic generation of the appropriate and product in the

medium (John, 2007).

15
 CHAPTER FOUR

4.0 Result

The samples obtained for this research were labelled using codes such as A

which represent Nahuta wells B represent Barracks wells C, represent Unguwan

Lamba wells and D represent Kuwet wells respectively in Potiskum town for

the purpose of identification. As shown in the table below.

Table 1. Showing the Sampling Stations

S/NO SAMPLE CODES SAMPLE Station

1 A Nahuita Wells
2 B Barracks Wells
3 C Unguwan Lamba Wells
4 D Kuwet Wells

16
 Table 2: Bacterial Load (Total Viable Laint) Cfu/Mc

S/NO SAMPLE CODES TOTAL VIABLE COUNT

1 A 3.0 x 104
2 B 2.6 x 104
3 C 2.0 x 104
4 D 1.9 x 104
WHO Standards 1.0 x 102
NAFDAC Standards 1.0 x 102

Table 2 showed the bacterial load (total viable count) of the areas under

investigation. As the total viable count in the table 2 above all the samples were

not in compliance WHO and NAFDAC standard.

Table 3: Total Coliform count Cfu / Ml

S/NO SAMPLE CODES COLIFORM COUNT (Cfc/Ml)

1 A 2.9 x 104
2 B 1.6 x 103
3 C 1.2 x 103
4 D 1.1 x 103
1.0 x 102
WHO Standards 1.0 x 102
NAFDAC Standards

Table 3 showed that the water samples were very closed to the standard limit

stipulated by WHO, and NAFDAC while other sample is not in agreement with

standard limit.

17
Table 4: Biochemical Characteristics of Bacterilsolates from the Sample

S/NO SAMPLE CODES CAT COT IND CIT

1 A +ve -ve +ve -ve
2 B +ve -ve +ve -ve
3 C +ve -ve +ve -ve
4 D +ve -ve +ve -ve

Key:

CAT : Represent Catalase Test

COT : Coagulase Test
IND : Indole Test
CIT : Citrase Test

Table COAGULASE TEST

S/NO SAMPLE COT PROBABLE ORG/
1 A -ve Staphylococus Epidermidis
2 B - ve Staphylococus Epidermidis
3 C +ve Staphylococus Aureas
4 D -ve Staphylococus Epidermidis

CATALASE TESA
S/NO SAMPLE COT PROBABLE ORG/
1 A +ve Staphylococus Spp
2 B +ve Staphylococus Spp
3 C +ve Staphylococus Spp
4 D +ve Staphylococus Spp

INDOLE TEST
S/NO SAMPLE COT PROBABLE ORG/
1 A +ve Escherichia Coli
2 B +ve Escherichia Coli
18
3 C +ve Escherichia Coli
4 D +ve Escherichia Coli

CITRASE TEST
S/NO SAMPLE COT PROBABLE ORG/
1 A -ve Escherichia Coli
2 B -ve Escherichia Coli
3 C -ve Escherichia Coli
4 D -ve Escherichia Coli

Positive (+ve) = indicates the presence of an organism in the sample.

Negative (-ve) = indicates the absence of an organism in the sample.

Table 4 above showed the various types of lacterial Spp – isolates by different

biochemical characterization test using a coagulase test staphylococcus

epidemidis was found negative whereas staphylococcus aureus was found

positive by catalase test the staphylococcus specie was found positive and using

the indole test the E. Coli wad found positive and finally by citrase test the E.

Coli was found negative inclusively.

19
 CHAPTER FIVE

5.0 Discussion

Faecal pollution of drinking water introduces a variety of intestinal pathogens

(WHO, 2012) Accordingly, the total coliforms counts of all the water samples

were general high, the exceeded the WHO and (NSDWQ) standards limits of

1.0 x 102 coliform count for drinking water which was also adopted by

NAFDAC (WHO, 2012 and NSDWQ 2017). The high coliform counts vividly

indicates the water sources are faecally contaminated (EPA 2003, Osuinde and

Enuezie, 1999). None of the water samples competes with WHO and NSDQW

standard for coliform and EW. Coli in drinking water. According to WHO and

NSDQW standard every water samples that has coliform must be analyzed for

either faecal coliforms or E. Coli (NSDQW 2007 and WHO, 2012), with a view

of ascertaining contamination with human or animal waste and possibly

20
pathogenic bacteria or organism such as giardia and cryptosporidium may be

present (Shittu et al 2008 and Epa 2003). There is evidence of coliform

contamination in all the samples. The total viable counts and the total coliform

count are in complies with WHO and NSDQW standard of 1.0 x 10 2 Cfu/Ml.

(NSDQW, 2007 and WHO, 2012). This findings is not surprising considering

the population density and close proximity of available water sources to pit

latrines. The sewage could seep slowly into underground water thereby

polluting it. Also long-term usage of boreholes may lead to deterioration of the

water quality because pipelines may be corroded with Radom cracks and most

cases clogged with sediment (One Mano and Otun, 2008).

This will allow the passage of in organic metals and bacteria. The implication

of this findings is the possibility of the presence of pathogens that may cause

acute intestinal illness with are generally considered discomfort to health and

could become fatal for some susceptible groups (such as infants, elderly and

those who are sick) (Addo et al 2009, Olowe et all 2005 NSDWQ, 2007)

Generally, underground water is offer considered as purest form of water

(Shittu et, al 2008), although its vulnerability to contamination could be due to

improper contruction, animal waste, proximity to toilet facilities, sewage,

refuge dumpsite and various human activities surrounding it (Bilton 1994 and

Shittu et al 2008). The high number of Salmonella and Shigella Spp. In the

21
samples is not is agreement with WHO and NSDWQ water standard for

drinking use which states that these pathogenic organism must not be present in

the water, because they are of public health significance, having been associated

with gastrointestinal infection: diarrgea, dysentery, typhoid fever and other

forms of infection (WHO, 2012 and NSDWQ, 2017).

Opportunistic pathogens such as E. Coli Spp staphylococcus Spp and proteus

Spp were also identified which are known to account for up to 50% of

nosocomial infections that are of public health significance (Obi and Okocha,

2007) pseudomonas and klebsiella are know to produce enterotoxin (Bannet

and Lancette 1992). Proteus Spp belongs to the intestinal flors but is also

widety distributed in soil and water (Schlegal 2003).

According to World Health Organization (2012) and Nigerian standard of

drinking water quality (2017) all water use for drinking should contain zero E.

Coli.

5.2 Conclusion and Recommendation

The principal aims of monitoring drinking water are not prevent the spread of

water borne diseases and to protect the public health. The importance of access

to good quality water cannot be over emphasized. Increase in population in

Potiskum with rise in human activity pose a great pressure on provision of safe

22
drinking water. This necessitates large number of people to consume water

form non-standard source like wash borehole and water from vendors which

constitute a major problem due to close proximity of such sources of pit

latrines. Looking at the result discussed, it is evident that the water sources of

Potiskum metropolis is contaminated and does not comply with the guidelines

for drinking water specify by the world health organization (WHO), Nigerian

standard of drinking water quality (DSNWQ), National Agency for Food and

Drug Administration and Control (NAFDAC) and many other authorities

concern with the quantity of drinking water.

Therefore, it can be concluded that the water might pose threat to the health of

consumers. It is recommended that the appropriate authorities are made aware

of this problem and that the sanitary services in Potiskum and Yobe State in

general took proactive measures. The study also showed high level of faecal

coliform and other pathogens include E. Coli, salmonella Spp, shigella Spp,

proteus Spp, pseudomonas Spp and klebsiells Spp, thus this water is not

suitable for drinking purposes, because the count were found to exceeded the

guidelines. It is evident that this water could pose a risk of infection to the

consumers ingesting it without proper treatment. There is need to create

awareness about present situation of sources of drinking water and the necessity

for further treatment by simple boiling by consumers before it can be used for

23
both drinking and domestic purposes. I would like to further recommend the

following important point. Proper sanitation, survey, design and implementation

of water and/or sanitation projects, regular disinfections, maintenances and

supervisions of water sources and regular bacteriological assessment of all

water sources for drinking should be planned and conducted.

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