ABSTRACT
This study aimed to isolate, identify, and
enumerate bacteria present in the oral cavity of students from Michael Okpara
University of Agriculture, Umudike. The oral microbiota, comprising a complex
community of microorganisms, plays a crucial role in maintaining oral health
and influencing disease outcomes. Samples were collected using sterile swabs
from students before and after brushing their teeth, providing insight into
bacterial load variations. The research sought to isolate and identify
bacterial species, quantify their presence, and examine their roles in oral
health and disease.
Bacterial isolation was performed using the pour
plate method, followed by enumeration through colony counts on Nutrient and
MacConkey agar. Serial dilutions were prepared, and bacterial colonies were
cultured and identified using biochemical tests and Gram-staining techniques. A
variety of bacteria were isolated, including Streptococcus spp., Staphylococcus
spp., Veillonella spp., Lactobacillus spp., and Fusospirochetes
spp. These results align with previous studies, indicating the oral
cavity's diverse microbial population.
The study revealed that Streptococcus spp.
was the most prevalent bacterium, playing a critical role in the formation of
biofilms and contributing to dental caries. Lactobacillus spp. was
identified as part of the normal flora, known for creating an acidic
environment that inhibits the growth of other bacteria. Staphylococcus spp.,
typically transient in the oral cavity, was associated with periodontal
disease. Other bacteria, such as Veillonella spp., were found to
thrive in acidic environments and may slow the progression of dental caries.
The findings highlight the dynamic balance of
oral microbiota, where some bacteria protect against disease, while others can
promote infection under certain conditions, such as poor oral hygiene.
Bacterial counts were significantly reduced after brushing, demonstrating the
importance of oral hygiene in controlling bacterial growth. The study
emphasizes the need for maintaining proper oral health practices to prevent
infections and periodontal diseases.
TABLE OF CONTENTS
CHAPTER
ONE
1.0 Introduction
1.1 Biofilm
and Oral Microbiota
1.2 Justification
of Study
1.3 General
Aims
1.4 Specific
Aims
CHAPTER
TWO
LITERATURE
REVIEW
2.0 The
Mouth As A Microbial Habitat
2.1
Heterogeneity of Oral Microbiota
2.2
Development of Resident
Microflora
2.3 Characteristics
Interactions between Oral Microorganisms.
2.4.
Cell-Cell Communication
2.5
Oral Bacterial
2.5.1 Streptococci
2.5.2 Lactobacillus
2.5.3 Actinobacillus Actinomycetemcomitans
2.5.4
Veillonella
2.5.5
Fusospirochetes
2.6
The Oral Microbiota and Disease
2.7 What
Can We Do To Minimize Disease Caused By The Oral Microbiota?
2.8
Roles of Resident Microbiota in
the Oral Cavity
CHAPTER
THREE
3.0
MATERIALS AND METHOD
3.1
Study Area
3.2
Collection of Sample
3.3
Media and Media Preparation
3.4
Isolation and Enumeration of
Bacterial from Oral Cavity
3.5
Identification of Bacteria
Isolates
3.5.1
Gram-Staining
3.5.2
Biochemical Reaction Tests
3.5.2.1
Coagulase test
3.5.2.2
Citrate test
3.5.2.3
Oxidase test
3.5.2.4 Motility
test
CHAPTER
FOUR
4.0 RESULTS
CHAPTER
FIVE
5.0
Discussion and Conclusion
5.1 Discussion
5.2
Conclusion
References
LIST OF TABLES
Table 4.1 : Microbial
(bacterial) count of oral cavity.
Table 4.2: Bacteria isolated from oral cavity and their percentage
occurrence
Table 4.3:
Biochemical test reaction
CHAPTER ONE
1.0
INTRODUCTION
The human oral cavity is
a diverse environment with hard and soft tissues comprising a total area of
215cm2 bathed in saliva (Eleni Kanasi, 2008). The oral environment
is thus optimal for their architecture and characteristics to biofilm in nature
(Auschill et al., 2001). Specific to
oral biofilms is the ability of sessile oral bacterial cells to tolerate short-term
abundance of external nutrient supply and to withstand nutrient restraint.
Nutrient deprived biofilm cells after reactivation have shown to exhibit low
reactivity invitro, suggesting that slower reactivation of these cells might be
a survival strategy (Chavez de paz et al.,
2008).
The oral cavity of
healthy individuals contains hundreds of different bacterial, viral and fungal
species. Many of these can associate to form biofilm which are resistant to
mechanical stress and antibiotic treatment. It has been a long journey from the
beginning to the study of oral microbiota consisting of an unknown number of
bacteria. But not all species residing in the oral microbiota have been
identified yet and an estimated 750 different species are anticipated (Jenkinson
and Lamont, 2005;Paster et al.,2006).
Most of these micro
organisms exist in our oral cavity in a symbiotic capacity maintaining
relationships with the hosts that are based on mutual benefits (los Alamos
National Library, 2009). Not only do they not cause harm, but also the
commensal populations may keep pathogenic species in check by not allowing them
to adhere to mucosal surfaces. Numer4oyus factors impede the isolation of this
vast number of species. First and foremost, many of the species are not cultural
with today’s laboratory technologies, and genomic similarities do not allow for
organismal determination based on short read lengths. Because of the
limitations, researchers have begun by identifying with the communities of
healthy mouth: Streptococcus, Actinomyces,
Staphylococcus, Veillonella, Neisseria, Eubacteria, Leptotrichia, Fusobacterium
etc.(Jenkinson and Lamot 2005; Wilson, 2005).
1.1 BIOFILM
AND ORAL MICROBIOTA
Some
bacterial invaders overcome the immune response by farming mixed biofilms
consisting of commensals and potential pathogens to cover fly hide within the
host (costerten, 2007). The formation of biofilm may occur on a wide variety of
surfaces in the oral cavity. Thus epithelial cells, saliva-coated enamels,
dental surfaces, primary colonizing bacteria and orthodontics all create
suitable environments for the establishment of mixed species biofilms (Jekinson
and Lamont, 2005; Marsh 2006). Bacteria cells in oral biofilms interact by
various recognized ways including co-aggregation ( Kolenbrander et al.,2006), metabolic exchange,
cell-cell communication ( Li et al., 2006)
and exchange of genetic material ( Robert et
al., 2001). These mechanisms benefits bacterial survival and can make
dental biofilms difficult therapeutic targets in dental diseases. At the
beginning of biofilm formation, initial surface attachment occurs by the primary
colonizers, resulting in disposition of a microbial monolayer.
The
primary colonizers in the oral microbiota for both mucosal and tooth surfaces
are usually streptococci which make up approximately 80% of early biofilms (Maria
Avila et al., 2009).
The
oral microbiota also faces challenges that are not experienced by other
microbiotas. The host has the option to maintain good personal hygiene, the cleansing
mechanism especially deliberate oral hygiene can alter the natural succession
that would otherwise define the climax community. This contributes to the
quantity and composition of mixed species biofilm in the oral cavity. In
response to eating, salivating, tooth brushing, tongue movement, flossing, and
other agitation, the oral microbial communities have evolved the skills to
survive these inhibitory practices.
Although biofilms are
required for health of the oral cavity, biofilms are also known to contain pathogens
(Ruby and Barbeau, 2002). Periodontal diseases such as chronic gingivitis and
periodontitis can result from an increase in the crevice. The biofilms are
typically comprised mainly of Gram-positive facultative anaerobes, but in the
absence of proper hygiene, the percentage of Gram-negative species in the
biofilms increases, contributing to periodontal inflammation (Ruby and Barbeau,
2002).
1.2 JUSTIFICATION OF STUDY
With our population aging
and living longer lives, health management and disease prevention become
increasingly important. Improved knowledge of the oral microbiota and their
impact/role on our health will contribute greatly to health of the society as a
whole. Also a better understanding of their dynamic complexity of oral microbiota
will contribute to the next-level in medical diagnostic tool. Ideally, this
study should also lead to providing the potential to manipulate the microbiota
to optimize personal health.
1.3 GENERAL AIMS
The objective of this
project research is to isolate, identify, enumerate and investigate the role of
the oral microbiota using microbiological and clinical methodology.
1.5
SPECIFIC AIMS
v To
isolate and identify bacteria from the oral cavity.
v To
enumerate oral microbiota.
v To
identify the roles of these organisms.
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