ABSTRACT
The present study aimed at comparing the microbiota of stool of healthy patients, and the microbiota of the diarrhoeic stool of patients with gastro intestinal tract infection. A total of 20 sample were used for the study, of which 7 were diagnosed of gastro intestinal enteritis in Abia state specialist hospital A.S.S.H Umuahia Aba road while the remaining were samples collected from ulcerative patient diagnosed of helicobacter pylori stool sample, while the remaining were 6 formed stool sample collected from healthy individuals in MOUAU. All the sample were cultured on salmonella-shigella agar, MacConckey agar for 24hours at 370C. Gram staining was done and biochemical test was also carried out in the pure culture out of 20 sample, 17 were positive. These were 6 salmonella spp, 7 Shigella spp. and Echerichia coli were bacteria pathogens responsible for diarrhea from the gastrointestinal patients. At a favourable condition, these microorganisms establish themselves in the host system, thereby posing a lot of clinical symptoms such as diarrhea, vomiting, abdominal cramp, headache, nausea etc which if not properly treated can lead to death. The best means of handling food borne infection and intoxication is by queuing into the measures of preventing it as detailed in chapter of this work.
TABLE OF CONTENTS
Title Pages
Title Page i
Certification ii
Dedication iii
Acknowledgements iv
Table of Contents v
List of Tables viii
Abstract ix
CHAPTER ONE: INTRODUCTION
1.1 Background of the Study 1
1.2 Justification of the Study 4
1.3 Objectives of the Study 4
1.3.1 General objectives 4
1.3.2 Specific objectives 4
CHAPTER TWO: LITERATURE REVIEW
2.1 Description of Different Bacteria 5
2.2 Bacillus Species 5
2.3 Salmonella
as Gastroenteric Pathogens 6
2.4 Staphylococci 8
2.5 Shigella
Gastroenteritis 10
2.5.1 Characterization of Salmonella Shigella 13
2.6 Antibiogram Study of Salmonella and Shigella 14
2.7 Clostridium
botulinum 15
2.8 Helicobacter
pylori 16
2.8.1 Pathogenesis of H. pylori 17
2.9 Human Faeces 17
2.9.1 Characteristics of human faeces 18
2.9.2 Stool analysis (stool sample) 21
2.9.3 Pathogens found in human faeces 22
2.10 Microorganisms 22
2.10.1 Functions of microbes in human gut 23
2.11 measures to be taken against bacterial
infection 25
CHAPTER THREE: MATERIALS AND METHODS
3.1 Description of Study Area 26
3.2 Sample Collection and Study Design 26
3.3 Study Design 26
3.4 Media and Media Preparation 27
3.4.1 Nutrient agar 27
3.4.2 Salmonella shigella agar 27
3.5 Identification
of Bacteria from the Stool of Healthy Individuals 28
3.5.1 Inoculation and incubation of the stool
sample 28
3.5.2 Identification of Bacteria from Diarrhoeic
Stool 28
3.5.3 Inoculation and incubation of stool sample 29
3.6 Characterization
of Bacteria Isolates from the Three Groups 29
3.5.1 Gram Staining 29
3.5.2 Triple sugar iron agar 30
3.5.3 Simmons citrate 30
3.5.4 Urea agar 31
3.5.5 Motility-indole-ornithine agar 31
3.5.6 Lysine iron agar 31
3.5.7 Methyl red test 32
3.5.8 Voges proskear test 32
3.6 Antimicrobial Susceptibility Test 32
CHAPTER FOUR: RESULTS
CHAPTER FIVE: DISCUSSION, CONCLUSION AND
RECOMMENDATIONS
5.1 Discussion 40
5.2 Conclusion 43
References 44
LIST OF TABLES
Table
Title Page
4.1: Distribution of bacterial
organism isolated from Helictober pylori
(ulcer)
patient in Abia State specialist Hospital Aba Road Umuahia 33
4.2:
Distribution of bacterial
organism isolated from gastrointestinal
tract
of (diarrhea) patients in Abia state
specialist hospital (ASSH) 34
4.3: Morphological Characteristics
of Bacterial Isolates
4.4: Distribution of bacterial
organism isolated from formed stool samples
of
healthy individual in MOUAU 36
4.5: Microbial Count, pH and
Temperature of the Feaces Samples 37
4.6:
Summary of the cultural
characteristic of bacteria isolated from the
stool sample 38
4.7:
Biochemical reaction of
the organism isolated from the stool samples 39
CHAPTER ONE
INTRODUCTION
1.1 BACKGROUND OF THE STUDY
The
bacteria found in human stool are representative of the bacteria that are present
in gastrointestinal tract. Certain bacteria and fungi called normal flora
inhabit every human being gastro intestinal (GI) tract. They play an important
role in the digestion of food and their presence keeps a check on the growth of
disease-causing bacteria (Bibiloni et al.,
2008).
Researchers
have identified a new link between the diversity of bacteria in human faeces
known as the human faecal microbiome.
The
GI tract is a complex and dynamic ecosystem containing a diverse collection of
microorganisms. These microorganisms are either resident members of the
intestinal microbiota or transient passengers in introduced from the
environment, for example by the regularly influx of microorganisms by the
intake of food (Flint and Duncan et al.,
2004).
The
human intestinal microbial population is comprised of at least a trillion
bacterial cells per gram of feaces. Most of these bacteria can be classified
into 400-800 individual species, the majority of which are only known by
sequence analysis (Reid and Greene, 2014).
This
gut microbial reaction is in close balance with the human host as it protects
against pathogens, detoxifies potential poison, produces energy and nutrients
by digestion of food and de novo synthesis, conditions and maintains mucosal
and systemic immunity and facilitates homeostasis.
Paradigm
shift is a phrase recently and frequently used among scientist when referring
to our new found mutualistic relationship with the microbes that inhibit the
human body. The bacteria found in human stool are representative in
gastrointestinal tract. Certain bacteria and fungi called normal flora inhibit
every human being gastro intestinal (GI) tract. They play an important role in
the digestion of food and their presence keeps a check on the growth of
diseases-causing bacteria. Probiotic are preparations which contain live
beneficial bacteria that when ingested have positive effects on the health of
the host.
Research
suggest that the ingestion of live bacteria can add to the, microbiota normally
found in the gastro intestinal tract. Human faeces are the faeces of human
digestive system including bacteria. They vary significantly in appearance i.e.
(size, colour, texture) according to the state of the digestive system, diet
and general health. The number of bacterial found in the human stool indicates
the level of bacteria present in the gastrointestinal tract thus, the GI tract
harbours many distinct niches, each containing a different microbial ecosystem
that varies according to the location within the GI tract. The understanding of
the composition of human faeces will guide us in the composition of the
microbial diversity of human stool sample.
Both
H. pylori and gut microbiota regulate
the host metabolism. Some clinical observations and animal studies support the
relationship of H. pylori infection and dysbiosis of gut microbiota to
metabolic disorders like insulin resistance and diabetes. However, how H.
pylori and gut microbiota interact with each other to regulate the host metabolism
remains unknown. Further studies are warranted to identify the H. pylori-microbiota-host metabolism
axis and to test if the eradication of H.
pylori or the modification of gut microbiota can control to treat human
metabolic disorders (Qin et al., 2012).
Diarrhea
is the passage of unusual loose or watery stools, usually at least three times
within 24 hour period. Prolonged diarrhea may lead to 24 excessive loss of
fluid, salt and nutrient in the faeces. The main cause of death from acute
diarrhea is dehydration, which result from loss of fluid and electrolyte in
stool. Another important cause of death is dysentery and under nutrition
(Sinclair et al., 2003).
Diarrhea
is an important cause of under nutrition because patients eat less during
diarrhea and their ability to absorb nutrient is reduced. Moreover, nutrient
requirement is increased as a result of infection (Sinclair et al., 2003). Risk factors that
predispose children to diarrhea include poor sanitation, poor social economic
status and malnutrition (Andu et al.,
2002).
Diarrhoeal
diseases are major attendance at health facilities , a common cause of
admission to many of the hospitals in the country and a significant and often
preventable cause of death the clinical symptoms of diarrhea include the
passage of frequent loose or watery stool without visible blood (Bahal et al., 2001), vomiting may occur and
fever may be present. The most important cause of acute watery diarrhea is
young children in Nigeria include rotovirus enteroxigenic Escherichia
coli, shigella campylobacter jejuni
and cryptosporidia, vibro cholera, salmonella and enteropathogenic, Escherchia
coli (Bahal et al., 2001).
Another
clinical symptom of diarrhea is dysentery, whixh refers to diarrhea with
visible blood in faeces, the symptoms of which include anorexia, rapid weight
loss and damage to the intestinal mucosa by invasive bacteria. The organisms implicated in this
type of diarrhea include shigella, campylobacter jejuni, slamonella and very
rarely Entaemoeba histolytica (Bahal et
al., 2001).
Transmission
of agent that cause diarrhea are usually of the faecal oral route, which
include the ingestion of faecal oral route, which include the ingestion of
faecal contaminated water or food, person to person contact and direct contact
with infected faces. Host factors that increase susceptibility to diarrhea
include under nutrition current or recent measles and immune deficiency or
immunosuspension (Andu et al., 2002).
Diarrhea
disease is part of the social problem in Nigeria and in other developing
countries in the tropics. Diarrhea disease is a leading cause of morbidity and
mortality among young children is low income countries (Okolo et al., 2013). The diarrhea specifics in
children less than 5 years age in Africa has been estimated at about 106 per
100. Therefore, the prevalence is not is
not fully understood and the isolating techniques is rather too expensive
(Olowe et al., 2003).
1.2 JUSTIFICATION OF THE STUDY
The
justification of the study is anchored on the fact that millions of pathogens
causing gastro intestinal tract infection are found in human stool. This study
highlights this fact and recommends preventive measures for public health
interest.
1.3 OBJECTIVES OF THE STUDY
1.3.1 General objectives
The
general objective of the study is aimed at studying the microbial diversity in
human stool sample.
1.3.2 Specific objectives
The
Specific objectives of the study include:
i.
To isolate and
characterize bacteria form healthy human stool.
ii.
To isolate and
characterize bacteria from the stool of ulcer patients.
iii.
To isolate and
characterize bacteria from the stool of diarrhea patients.
To compare the
diversity of bacteria from the three
different stool sample.
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