Microbiology Oral Biology Dr.

Radwa Khalil

Classification of microorganism

microorganisms

cellular acellular

prokaryotes virus

eukaryotes prions

Prokaryotes
are simple, small cells, whereas eukaryotic cells are complex, large structured and are present in trillions
which can be single celled or multicellular. Prokaryotic cells do not have a well-defined nucleus but DNA
molecule is located in the cell, termed as nucleoid, whereas eukaryotic cells have a well-defined nucleus,
where genetic material is stored. Based on the structure and functions, cells are broadly classified as
Prokaryotic cell and Eukaryotic cell

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Prokaryotic Cells are the most primitive kind of cells and lack few features as compared to the
eukaryotic cell. Eukaryotic cells have evolved from prokaryotic cells only but contain different types of
organelles like Endoplasmic reticulum, Golgi body, Mitochondria etc, which are specific in their
functions. But features like growth, response, and most importantly giving birth to the young ones are
the commonly shared by all living organisms.

Definition of Prokaryotic Cells

Pro means ‘old,’ and karyon means ‘nucleus,’ So as the name suggest the history of the evolution of
prokaryotic cells is at least 3.5 billion years old, but they are still important to us in many aspects like
they are used in industries for fermentation (Lactobacillus, Streptococcus), for research work, etc. In
comparison to eukaryotic cells, they lack few organelles and are not advanced as eukaryotes.

Generalized structure of Prokaryotic cell consists of the following:

Glycocalyx: This layer function as a receptor, the adhesive also provide protection to the cell wall.

Nucleoid: It is the location of the genetic material (DNA), large DNA molecule is condensed into the
small packet.

Pilus: Hair like hollow attachment present on the surface of bacteria, and is used to transfers of DNA to
other cells during cell-cell adhesion.

Mesosomes: It is the extension of the cell membrane, unfolded into the cytoplasm their role is during
the cellular respiration.

Flagellum: Helps in movement, attached to the basal body of the cell.

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Cell Wall: It provides rigidity and support for the cell.

Fimbriae: Helps in attachment to the surface and other bacteria while mating. These are small hair-like
structure.

Inclusion/Granules: It helps in storage of carbohydrates, glycogen, phosphate, fats in the form of

particles which can be used when needed.

Ribosomes: Tiny particles which help in protein synthesis.

Cell membrane: Thin layer of protein and lipids, surrounds cytoplasm and regulate the flow of materials
inside and outside the cells.

Endospore: It helps cell in surviving during harsh conditions.

In terms of peptidoglycan present in the cell wall, prokaryotes can be divided into Gram-positive and
Gram -negative bacteria. The former contains a large amount of peptidoglycan in their cell wall while
the latter have the thin layer.

Eukaryotic Cells
Definition of Eukaryotic Cells

Eu means ‘new,’ and karyon means ‘nucleus,’ so these are the advanced type of cells found in plant,
animals, and fungi. Eukaryotic cells have a well-defined nucleus and different organelles to perform
different functions within the cell, though working is complex to understand.

This kind of cells are found in algae, fungi, protozoa, plants, and animals and can be single-celled,
colonial or multicellular. Among them, fungi and protists (algae and protozoa) are the major kingdoms.

The general structure of Eukaryotic cells contain:

Nucleus: Eukaryotic cells have a well-defined nucleus where DNA (genetic material) is stored, it helps in
the production of protein synthesis and ribosomes also. The chromosome is present inside the nucleus,

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which is surrounded by the nuclear envelope. It is a bi-lipid layer and controls the passage of ions and
molecules.

Cytoplasm: It is the location where other organelles are located, and other metabolic activities of the
cell also take place here. It consists of –

Mitochondria: It is called ‘the powerhouse of the cell,’ and is responsible for making ATP. Mitochondria
has its own DNA and ribosomes.

Chloroplast: These are found in algae and plants, it is one of the most important organelles in the plant
which helps in converting energy sunlight into chemical energy through photosynthesis. They resemble
mitochondria.

Golgi Apparatus: It consists of a stack of many flattened, disc-shaped sacs known as cisternae. The exact
nature of Golgi varies, but it helps in the packaging of materials and in secreting them.

Lysosomes and Vacuoles – The most important function of Endoplasmic reticulum and Golgi apparatus is
the synthesis of Lysosomes, which helps in digestion of intracellular molecules with the help of the
enzyme called hydrolase.

Vacuoles are the membrane-bound cavities containing fluid as well as solid materials, and they engulf
materials through endocytosis.

Endoplasmic Reticulum: It transport lipids, proteins, and other materials through the cell. They are of
two types of smooth endoplasmic reticulum and rough endoplasmic reticulum.

Appendages: Cilia and Flagella are locomotory attachments, helps in the movement of a cell towards
positive stimuli. Cilia are shorter than flagella and numerous.

Surface structure: Glycocalyx is a kind of polysaccharide, and it is the outermost layer of the cell which
helps in cell adherence, protection and in receiving signals from other cells.

Cell Wall: Cell Wall provides shapes, rigidity, and support to the cell. Compositions of the cell wall may
vary of different organisms but which can be of either cellulose, pectin, chitin or peptidoglycan.

Cytoplasmic Membrane/Plasma Membrane: It is a thin semipermeable, surrounding the cytoplasm, it

acts as the barrier of the cell which regulates entry and exit of the substances inside and outside the cell.
This layer is made up of two layers of phospholipids embedded with proteins. In Plant cell, this layer is
present below the cell wall whereas in the Animal cell it is the outermost layer.

Ribosomes: Though small in size but are present in numbers, they help in protein synthesis. Eukaryotes
have 80S ribosomes which are further divided into two subunits which are 40S and 60S (S stands for
Sedverg unit).

Cytoskeleton: It is supporting framework of the cells, which is of two types Microtubules and
Microfilaments. Microtubules have a diameter of about 24 nanometers (nm), made up of a protein
called tubulin, while Microfilaments has a diameter of 6nm, made of the protein called actin.
Microtubules are the largest filament and Microfilament the smallest one.

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Key Difference Between Prokaryotic Cells and Eukaryotic Cells

Following are the substantial difference between Prokaryotic Cells and Eukaryotic Cell:

Prokaryotic cells are the primitive kind of cell, whose size varies from 0.5-3µm, they are generally found
in single-cell organisms, while Eukaryotic cells are the modified cell structure containing different
components in it, their size varies from 2-100µm, they are found in multicellular organisms.

Organelles like mitochondria, ribosomes, Golgi body, endoplasmic reticulum, cell wall, chloroplast, etc.
are absent in prokaryotic cells, while these organelles are found in eukaryotic organisms. Though cell
wall and chloroplast are not found in the animal cell, it is present in the green plant cell, few bacteria,
and algae.

The main difference between Prokaryotic cells and the Eukaryotic cell is the nucleus, which is not well
defined in prokaryotes whereas it is well structured, compartmentalized and functional in eukaryotes.

Cell organelles are present which are membrane-bound and have individual functions in eukaryotic
cells; many organelles are absent in prokaryotic cells.

In prokaryotes, the cell division takes place through conjugation, transformation, transduction but in
eukaryotes, it is through the process of cell division.

The process of transcription and translation occurs together, and there is a single origin of replication in
the prokaryotic cell. On the other hand, there are multiple origins of replication and transcription occurs
in nucleus and translation in the cytosol.

Genetic Material (DNA) is circular and double-stranded in Prokaryotes, but in Eukaryotes, it is linear and
double-stranded.

Prokaryotes reproduce asexually; commonly Prokaryotes have a sexual mode of reproduction.

Prokaryotes are the simplest, smallest and most abundantly found cells on earth; Eukaryotes are larger
and complex cells.

Virus
Viruses are made up of a protein coat, or capsid, which encloses either DNA or RNA, the genetic code for
the virus.

they lack cell structure and they cannot reproduce outside a host cell. These two are characteristics that
scientists use to define life.

Not living cells

Once inside the body cell, viral DNA or RNA can be replicated and thousands of new viral particles are
produced. The host cell dies and the new viral particles infect more host cells.

Influenza, Chickenpox (varicella), Herpes, and HIV/AIDS

Prions
Prions consist only of protein

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“Mad cow disease” (bovine spongiform encephalopathy)

Both virus and prions are too small to be seen by light microscope

Not considered living organisms

Don’t replicate outside the host cells

They are not part of the body flora

The flora of the body only consists of Bactria and fungi ….that is why we will study the bacteria in more
details

Bacterial classification
Morphology
Shape and size

The shape of a bacterium is determined by its rigid cell wall.

Bacteria are classified by shape into three basic groups

1. cocci (spherical)

2. bacilli (rod-shaped)

3. spirochaetes (helical).

Some bacteria with variable shapes, appearing both as coccal and bacillary forms, are called
pleomorphic (pleo: many; morphic: shaped) in appearance.

The size of bacteria ranges from about 0.2 to 5 μm. The smallest bacteria approximate the size of the
largest viruses (poxviruses), whereas the longest bacilli attain the same length as some yeasts and
human red blood cells (7 μm).

Arrangement
Bacteria, whichever shape they may be, arrange themselves (usually according to the plane of
successive cell division) as pairs (diplococci), chains (streptococci), grape-like clusters (staphylococci) or
as angled pairs or palisades (corynebacteria).

Gram-staining characteristics
In clinical microbiology, bacteria can be classified into two major subgroups according to the staining
characteristics of their cell walls. The stain used, called the Gram stain (first developed by a Danish
physician, Christian Gram), divides the bacteria into Gram-positive (purple) and Gram-negative (pink)
groups. The Gram-staining property of bacteria is useful both for their identification and in the therapy
of bacterial infections because, in general, Gram-positive bacteria are more susceptible to penicillins
than Gram-negative bacteria.

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Structure
Bacteria have a rigid cell wall protecting a fluid protoplast comprising a cytoplasmic membrane and a
variety of other components

Structures external to the cell wall

Flagella
Flagella are whip-like filaments that act as propellers and guide the bacteria towards nutritional and
other sources. The filaments are composed of many subunits of a single protein, flagellin. Flagella may
be located at one end (monotrichous, a single flagellum; lophotrichous, many flagella) or all over the
outer surface (peritrichous). Many bacilli (rods) have flagella, but most cocci do not and are therefore
non-motile. Spirochaetes move by using a flagellum-like structure called the axial filament, which wraps
around the cell to produce an undulating motion.

Fimbriae and pili

Fimbriae and pili are fine, hair-like filaments, shorter than flagella, that extend from the cell surface. Pili,
found mainly on Gram-negative organisms, are composed of subunits of a protein, pilin, and mediate
the adhesion of bacteria to receptors on the human cell surface – a necessary first step in the initiation
of infection. A specialized type of pilus, the sex pilus, forms the attachment between the male (donor)
and the female (recipient) bacteria during conjugation, when genes are transferred from one bacterium
to another.

Glycocalyx (slime layer)

The glycocalyx is a polysaccharide coating that covers the outer surfaces of many bacteria and allows the
bacteria to adhere firmly to various structures, e.g. oral mucosa, teeth, heart valves and catheters, and
contribute to the formation of biofilms. This is especially true in the case of Streptococcus mutans, a
major cariogenic organism, which has the ability to produce vast quantities of extracellular
polysaccharide in the presence of dietary sugars such as sucrose.

Capsule
An amorphous, gelatinous layer (usually more substantial than the glycocalyx) surrounds the entire
bacterium; it is composed of polysaccharide, and sometimes protein (e.g. anthrax bacillus).

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Cell wall
The cell wall confers rigidity upon the bacterial cell. It is a multilayered structure outside the cytoplasmic
membrane. It is porous and permeable to substances of low molecular weight.

Bacterial physiology
Growth
Bacteria, like all living organisms, require nutrients for metabolic purposes and for cell division, and
grow best in an environment that satisfies these requirements.

Chemically, bacteria are made up of polysaccharide, protein, lipid, nucleic acid and peptidoglycan, all of
which must be manufactured for successful growth.

Nutritional requirements
Oxygen and hydrogen
Both oxygen and hydrogen are obtained from water; hence, water is essential for bacterial growth. In
addition, the correct oxygen tension is necessary for balanced growth.

While the growth of aerobic bacteria is limited by availability of oxygen, anaerobic bacteria may be
inhibited by low oxygen tension.

Carbon
Carbon is obtained by bacteria in two main ways:

1. Autotrophs, which are free-living, non-parasitic bacteria, use carbon dioxide as the carbon source.

2. Heterotrophs, which are parasitic bacteria, utilize complex organic substances such as sugars as their

source of carbon dioxide and energy.

Inorganic ions
Nitrogen, sulphur, phosphate, magnesium, potassium and a number of trace elements are required for
bacterial growth.

Organic nutrients
Organic nutrients are essential in different amounts, depending on the bacterial species:

• Carbohydrates are used as an energy source and as an initial substrate for biosynthesis of many
substances.

• Amino acids are crucial for growth of some bacteria.

• Vitamins, purines and pyrimidines in trace amounts are needed for growth.

Reproduction
Bacteria reproduce by a process called binary fission, in which a parent cell divides to form a progeny of
two cells.

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Other factors that affect the doubling time include the amount of nutrients, the temperature and the pH
of the environment.

Bacterial growth cycle

The growth cycle of a bacterium has four main phases

1. Lag phase: may last for a few minutes or for many hours as bacteria do not divide immediately
but undergo a period of adaptation with vigorous metabolic activity.
Bacteria are first introduced into an environment or media
Bacteria are “checking out” their surroundings
Bacteria adjusting to new environment
Cells are very active metabolically
Number of cells changes very little
1 hour to several days
2. Log (logarithmic, exponential) phase: rapid cell division occurs, determined by the
environmental conditions.
Each time binary fission occurs = doubling in numbers of bacteria
Microbes are sensitive to adverse conditions- antibiotics & anti-microbial agents
3. Stationary phase: this is reached when nutrient depletion or toxic products cause growth to
slow until

the number of new cells produced balances the number of cells that die. The bacteria have now
achieved their maximal cell density or yield.

Total number of viable cells remains constant (number of new cells = number of dying cells)

Endospores would form now

Cells begin to encounter environmental stress-lack of nutrients & water & oxygen, not enough space,
metabolic wastes, change in pH

4. Decline or death phase: this is marked by a decline in the number of live bacteria.
Growth stops
Dead cells more than number viable cells
Death rate > rate of reproduction
Due to limiting factors in the environment

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Growth regulation
Bacterial growth is essentially regulated by the nutritional environment. However, both intracellular and
extracellular regulatory events can modify the growth rate. Intracellular factors include:

• end product inhibition: the first enzyme in a

metabolic pathway is inhibited by the end product of

that pathway

• catabolite repression: enzyme synthesis is inhibited by

catabolites.

Extracellular factors that modify bacterial growth are:

• Temperature: the optimum is required for efficient activity of many bacterial enzymes, although
bacteria can grow in a wide range of temperatures. Accordingly,

bacteria can be classified as:

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 mesophiles, which grow well between 25 and 40°C,

comprising most medically important bacteria (that grow best at body temperature)

 thermophiles, which grow between 55 and 80°C

(Thermus aquaticus, for instance, grows in hot springs and its enzymes such as Taq polymerase are
therefore heat resistant, a fact exploited by molecular biologists in the polymerase chain reaction (PCR)

 psychrophiles, which grow at temperatures below 20°C.

 pH: the hydrogen ion concentration of the environment should be around pH 7.2–7.4 (i.e.
physiological pH) for optimal bacterial growth.
However, some bacteria (for example, lactobacilli) have evolved to exploit ecological niches,
such as carious cavities where the pH may be as low as 5.0.

Aerobic and anaerobic growth

A good supply of oxygen enhances the metabolism and growth of most bacteria. The oxygen acts as the
hydrogen acceptor in the final steps of energy production and generates two molecules: hydrogen
peroxide (H2O2) and the free radical superoxide (O2). Both of these are toxic and need to be destroyed.
Two enzymes are used by bacteria to dispose of them: the first is superoxide dismutase, which catalyses

the reaction:

2O2 + 2H+ -H2O2 +O2

and the second is catalase, which converts hydrogen peroxide

to water and oxygen:

2H2O2 2H2O + O2

Bacteria can therefore be classified according to their ability to live in an oxygen-replete or an oxygen-
free environment This has important practical implications, as clinical specimens must be incubated in
the laboratory under appropriate gaseous conditions for the pathogenic bacteria to grow. Thus, bacteria
can be classified as follows:

• obligate (strict) aerobes, which require oxygen to grow because their adenosine triphosphate (ATP)-
generating system is dependent on oxygen as the hydrogen acceptor (e.g. M. tuberculosis)

• facultative anaerobes, which use oxygen to generate energy by respiration if it is present, but can use
the fermentation pathway to synthesize ATP in the absence of sufficient oxygen (e.g. oral bacteria such
as mutans streptococci, E. coli)

• obligate (strict) anaerobes, which cannot grow in the presence of oxygen because they lack either
superoxide dismutase or catalase, or both (e.g. Porphyromonas gingivalis)

• microaerophiles, that grow best at a low oxygen concentration (e.g. Campylobacter fetus).

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The virulence factors of microorganisms

Virulence: is referred to the ability of microbial strains to produce disease.
•Virulence Factor – a pathogen-produced substance that promotes the establishment and
maintenance of disease

•Invasiveness: the ability of organism to spread in a host tissue after establishing infection.

•Toxigenicity: the ability of a microorganism to produce a toxin that contributes to the development of
disease.

The various virulence factors of microorganisms:

1)Movement Pili/Fimbriae

2)Glycocalyx 1.Slime layer

(cover) 2.Capsules
3) Enzymes 1.Collagenase
2.Coagulase
3.Streptokinase
Hemolysins 1.Lecithinase /
Phospholipase
2.Streptolysin O
3.Leukocidins
5) Toxins 1.Endotoxin Endotoxins are the cell wall lipopolysaccharides of Gram
2.Exotoxin negativebacteria (both cocci and bacilli) and are not actively
released from the cell. (Note: thus, by definition, Gram
positive organisms do not possess endotoxins.) Endotoxins
cause fever, shock and other generalized symptoms
exotoxins
Both Gram-positive and Gram-negative bacteria
secrete exotoxins, whereas endotoxin is an integral
component of the cell wall of Gram-negative organisms. are
highly toxic

The pathogenic properties of microorganism

1)Adherence

2)Invasion of host tissues

3)Evasion of host defense

4)Tissue damage

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1. Adherence
Pathogen must adhere to host cells to establish infection

•Some microbes produce materials or structures that allow them to adhere (stick) to membranes or
surfaces, and thus escape defenses:

(a) Pili/ Fimbriae

(b) Glycocalyx (slime layer)

(c) Capsules

(d) Spikes – Viral envelopes of some viruses, Influenza a, H5N1

2. Invasion and Evasion of host defense

•Microbes penetrate host barriers by toxin and enzyme production and then multiply in tissues

•The major antiphagocytic contributing factor:

1.Capsule

2.Pili

3.Surface proteins – Proteins prevent phagocytosis (leukostatin, leukocydins of Staph and Strep)

4.Evade immune response - Genetic variability occurs and the result is that antibodies lose effectiveness
quickly.degrade their respective intercellular substances, allowing easy spread of bacteria through
tissues, and are especially important in skin infections caused by Streptococcus pyogenes

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3. Damage of host tissues

•Bacteria will first adhere on the tissue and causes tissue damage

•Damage can be direct result of pathogen, such as toxin production, or indirect via immune response

•The major tissue damage contributing factors :

(a) Enzymes

(b) Bacterial toxins

(c) Hemolysins

Normal flora:
Normal flora: the population of microorganisms that inhabit the skin and mucous membranes of healthy
normal persons

•All body surfaces possess a rich normal bacterial flora, especially the mouth, nose, gingival crevice,
large bowel and skin

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•Shifts in the normal microbiota or stimulation of inflammation by these commensals may cause
diseases such as bacterial vaginosis, periodontitis, and inflammatory bowel disease.

Microflora usually is non-pathogenic and form an integral part of the host.

•The “normal microbiota” provides

•A first line of defence against microbial pathogens.

•Assist in digestion / synthesis of vitamins

•Play a role in toxin degradation,

•Contribute to maturation of the immune system.

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How can the oral flora lead to disease ?

Transient Microbiota
•Microorganisms that survive in the body only for a short period of time.

•Transient bacteria can be either benign or pathogenic.

•Transients are unable to remain in the body for extended periods of time due to:

•Competition from resident microbes;

•Elimination by the body’s immune system;

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•Physical or chemical changes within the body that discourage the growth of transient microbes.

Resident Microbiota
•Skin and mucous membranes, are constantly in contact with environmental organisms and become
readily colonized by various microbial species;

•The resident microbiota prevents colonization of pathogens on the mucous membranes and skin
(Bacterial interference)

•These microbes have a commensal relationship with their host, meaning that they do not cause harm
while they benefit from feeding on the cellular waste & dead cells of the host's body.

Endogenous microflora
•Bacterial flora is endogenous bacteria, which is defined as bacteria that naturally reside in a closed
system.

Exogenous microflora
•Exogenous bacteria are microorganisms introduced to closed biological systems from the external
world.

Opportunistic microbes
•Microorganisms that take advantage of certain opportunities to cause disease. They are frequently
members of the body’s normal flora

•Opportunistic microbe is only able to cause disease:

•In compromised individuals when their defenses are weak, as the microbes will take opportunity by the
reduced host defenses

•When the balance of normal microbes is disrupted, for example when a person takes broad spectrum
antibiotics, microbes that are normally crowded out by resident microbes have an opportunity to take
over.

•Disease can result when normal flora are traumatically introduced to an area of the body that they do
not normally occur in

Microbes & human relationship

•There are at least 3 types of relationships based on the quality of the relationship for each member of
the symbiotic association:

1.Mutualism

2.Commensalism

3.Parasitism

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Mutualism
•Mutualism, association between organisms of two different species in which each benefits.

•One classic mutualistic association is that of the lactic acid bacteria that live on the vaginal epithelium
of woman

•The bacteria are provided habitat with a constant temperature & supply of nutrients (glycogen) in
exchange for the production of lactic acid - protects the vagina from colonization & disease caused by
yeast & other potentially harmful microbes

Commensalism
•One organism benefits but the other is neutral (there is no harm or benefit)

•Staphylococcus epidermidis a consistent inhabitant of the skin of humans

•Staphylococcus epidermidis produces lactic acid that protects the skin from colonization by harmful
microbes that are less acid tolerant

Parasitism
•Parasitism is when one species in a relationship benefits and the other is harmed.

•A parasite that causes disease in its host is called a pathogen.

•Some parasitic bacteria live as normal flora of humans while waiting for an opportunity to cause
disease.

Factors that determine normal flora

1.Temperature: highly constant temperature (warm- blood animals)

2.Moisture

3.Presence of certain nutrients: rich in organic nutrients & growth factors

4.Inhibitory substances

5.Relatively constant conditions of pH & osmotic pressure

6.Antibiotics,…..etc

Significance of Normal Flora

•The normal flora :

•Synthesize & excrete vitamins

•Prevent colonization by pathogens

•May antagonize other bacteria

•Stimulate the development of certain tissues

•Stimulate the production of natural antibodies

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Acquisition of the normal oral flora The origin of Oral Microflora

A few hours after birth, the oral cavity becomes colonized by the strains of bacteria that eventually form
the resident microflora, which establish a permanent population in the mouth;

•After that, as the individual is further exposed to external sources of bacteria, the biodiversity of the
oral cavity increases, to a point where stability is reached. This is termed the climax community

The source of oral microorganisms

•The source of these microorganisms mainly comes from saliva, although it also includes the food and
water consumed by the individual;

• vertical transmission :Majority of children obtain their resident microflora from their mothers, as they
often possess identical strains of bacteria; This is known as vertical transmission;

•Horizontal transmission also takes place as children interact with their peers, and later in life between
spouses and partners.

Periods of formation of the oral ecosystem

At birth:
•The oral cavity is composed only of the soft tissues of the lips, cheeks, tongue and palate, which are
kept moist by the secretions of the salivary glands.

•At birth the oral cavity is sterile but rapidly becomes colonized from the environment, particularly from
the mother in the first feeding.

•Streptococcus salivarius is dominant and may make up 98% of the total oral flora until the appearance
of the teeth (6 - 9 months in humans).

The neonatal period:

•Obtained from the mother and the surrounding area;

•Mostly aerobes;

•Candida albicans - 54%;

•Period before eruption:

•Facultative anaerobes and anaerobes;

•Streptococcus spp.- 90%;

•Actinomyces;

•Candida albicans.

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After eruption:
•The eruption of the teeth during the first year leads to colonization by Streptococcus mutans and S.
sanguis;

•They will persist as long as teeth remain;

•Other strains of streptococci adhere strongly to the gums and cheeks but not to the teeth.

Period after eruption:

•Anaerobes:

Actinomyces;

Lactobacillus;

Veillonella;

Fusiformis;

Spirochaetes;

Candida albicans.

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Oral Habitats
(1)Buccal mucosa (Lips, palate and cheek) 3)Teeth
-S. oralis, S. sanguis; -Streptococcus spp,
-Neisseria spp.; -Actinomyces spp,
-Haemophilus spp. ; -Veillonella spp,
-Veillonella spp. -Fusobacterium spp, and
-Prevotella spp
(2) Dorsum of tongue
-S. salivarius, S. mitis,
-Actinomyces spp.;
-Rothia spp,
-Nisseria spp,
-Veillonella spp.;
-Peptostreptococcus spp.
-Bacteroides spp.;
(4) Supragingival plaque (5) Subgingival plaque
-Streptococcus spp.; -P. gingivalis;
-Actinomyces spp.; -P. endodontalis;
-Veillonella spp., -Prevotella intermedia;
-Haemophilus spp., -Prevotella denticola
-Bacteroides spp.

The role of oral flora in systemic disease

•Recently, it has been recognized that plaque-related oral disease, especially periodontitis may alter the
course and pathogenesis of a number of systemic diseases.

•These include:

–Cardiovascular disease:

•Infective endocarditis

•Coronary heart disease

•Atherosclerosis

•Myocardial infection

•Stroke

–Bacterial pneumonia

–Diabetes mellitus

–Low birth weight babies

How can the oral infection cause systemic disease

Three mechanisms linking oral infections to secondary systemic disease have been proposed:

1.Metastatic infection

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2.Metastatic injury

3.Metastatic inflammation

What is metastasis:
the development of secondary malignant growths /infection at a distance from a primary site of
cancer/disease

Factors which influence the growth of microorganisms in the oral cavity

(1)Saliva
–Adsorption on the tooth surface forms a salivary pellicle, a film that facilitates bacteria adhesion.

–Acts as a primary source of food.

–Has a good buffering capacity, thus pH is maintained.

–Facilitate bacterial clearance from the mouth.

–Contribute in host defense by having :

• Non-specific defense factors (eg. lysozyme, lactoferrin, histatins).

• Specific defense factors (eg. immunoglobulins).

(2) Anatomical factors

–The structure & shape of the teeth

–Malalignment of teeth,

–Poor quality of restorations (eg. fillings, bridges)

–Non-keratinized sulcular epithelium

(3) Gingival crevicular fluid (GCF)

–Has a good buffering capacity.

–Acts as a primary source of food.

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–Facilitate bacterial clearance from the mouth.

–Providing specific and nonspecific defense factors.

–Phagocytosis: 95% of leucocytes in the crevicular fluid are neutrophils.

Factors which influence the growth of microorganisms in the oral cavity

(4) Microbial factors

–Competition for receptors for adhesion at colonizing sites.

–Production of metabolic end products and toxins that kill other bacteria of the same/different sp.

–Use of metabolic end products of other bacteria for nutritional purposes

(5) Environmental pH
–Many microbes require a neutral pH for growth (saliva has mean pH 6.7)

–However, the environment pH depends on the frequency of dietary carbohydrate intake.

(6) Antimicrobial therapy

–Mouth rinses with antimicrobial agents such as chlorhexidine.

(7) Diet & Nutrition

–Exogenus nutrient source : Food

– Endogenus nutrient source : Host and Microbes

(a) Host : saliva (amino acids, vitamins, glycoprotein) and crevicular fluid (proteins, glycoprotein)

(b) Microbes : intracellular polysaccharides (glycogen) and extracellular microbial products

Examples of Oral Diseases

The dental professional comes into contact with two of the most widespread of all human diseases -
dental caries and periodontal diseases

1- Dental Caries
Dental caries is a chronic infection of enamel or dentine in
which the microbial agents are members of the normal
commensal flora.

•Lesions result from the demineralization of enamel or

dentine by acids produced by plaque microorganisms as
they metabolize dietary carbohydrates.

Aetiological Factor of Dental Caries

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The main factors involved in dental caries are the tooth, saliva, supragingival plaque, the diet (especially
sucrose intake) and the time necessary for caries development

Microbiology of Dental Caries

• Strep. mutans early carious lesions of enamel

Lactobacilli dentinal caries

Actinomyces root caries

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2- Periodontal Disease
•A condition where tissues of the periodontium are infected by
bacteria causing inflammatory responses to the tissues

•Periodontal disease can cause a breakdown of the periodontium

resulting in loss of tissue attachment and destruction of the alveolar
bone.

Aetiological Factors of Periodontal Diseases

1.Host tissues

2.Microbes

3.Immune defense system

The main causative agent of Periodontal Disease is Porphyromonas

gingivalis

3- Dentoalveolar abscess
This common infection develops typically at the apices of the roots of teeth, following necrosis of the
pulp.

•Abscesses may arise de novo or may develop within a pre-existing granuloma.

The abscess may remain localized within the alveolar bone.

•Alternatively, the infection may burst through the alveolar bone

and into the soft tissues.

•This may result in intra- or extra-oral swelling or in potentially

dangerous spread of infection through fascial planes.

The main causative agent is Facultative anaerobes, particularly

those of the anginosus group of streptococci (S. anginosus, S.
intermedium and S. constellatus), are also found frequently

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4- Periodontal abscess
Periodontal abscesses usually occur in patients with established periodontal pockets.

•The periodontal abscess is of sudden onset.

•There is usually swelling, redness and tenderness of the overlying

gingiva.

Microorganisms play a role in periodontal abscess include anaerobic

Gram-negative rods, alpha haemolytic and anaerobic streptococci
together with others such as spirochaetes.

5- Ludwig's angina
•Ludwig's angina is a bilateral infection of the sublingual and sub-mandibular spaces.

•The infection often represents cellulitis of the fascial spaces, rather than true abscess formation.

•The key clinical features are a brawny oedema with elevation of the tongue, airway obstruction and

very little pus.

A wide range of organisms has been reported from these

infections, including staphylococci, streptococci and
enterobacteria. However, oral commensal bacteria, especially
anaerobic Gram-negative bacilli and anaerobic streptococci, are
most commonly isolated.

6- Osteomyelitis of the jaws

•Osteomyelitis of the jaws is rare infection.

•Osteomyelitis is defined as inflammation of the medullary cavity of

bone, but it usually spreads to involve the cortical bone and
periosteum as well.

•Following ischaemia, the infected bone becomes necrotic.

More modern studies indicate that members of the normal oral

flora, particularly anaerobic Gram-negative rods and anaerobic
streptococci, are usually the organisms of importance.

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Dental Plaque
Definition:
soft deposits that form the biofilm adhering to the tooth surface or other hard surfaces in the oral
cavity, including removable & fixed restorations

Dental Plaque is an invisible layer of microorganisms growing in a colony on the teeth. Plaque that
becomes calcified is referred to as calculus or tartar.

Plaque contains many types and a large number of closely packed bacteria.

Plaque begins to form immediately on a freshly polished tooth, but it takes approximately 21 days for
plaque to completely mature

If plaque is disturbed (brushed away) when it begins to form, it does not mature

places of plaque formation:

Plaque is found naturally on the tooth surface, Dental plaque always found at protected and stagnant
sites of tooth such as fissures, approximal regions between teeth, and the gingival crevice

Plaque is typically located at retention and stagnation areas along the gingival margin and between
teeth (approximal areas).

Plaque can accumulate beyond levels compatible with oral health, and this can lead to shifts in the
composition of the microflora and predispose sites to disease.

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Composition:

80% Water (of wet weight)

•50% intracellular

•30% extracellular

•20% Organic components (of wet weight)

•10% bacteria

•6% extracellular protein

•2% extracellular polysaccharide + inorganic components (mainly calcium & phosphate)

Types of Dental Plaque

Supragingival Plaque

Location:

•Above gingival margin Type of microbes:

•Aerobes

•Anaerobic Facultative

Subgingival Plaque

Location:

•Below gingival margin Type of microbes:

•Anaerobic Obligate

•Anaerobic Facultative

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Microbiology Oral Biology Dr. Radwa Khalil

Acquired pellicle:
a thin, microscopic coating of saliva materials. That covers the tooth surface within seconds of eruption
that is why it is exogenous origin (formed from substance outside the tooth),,it is removed by polishing
of the teeth

Materia Alba
“soft accumulations of bacteria and tissue cells; lack organized structure of dental plaque; easily
displaced by water spray”

Dental Calculus
‘mineralized dental plaque”

Steps of dental plaque formation/ Mechanism of Plaque Formation

1-Acquired pellicle formation:
DEFINATION : Which is a thin, microscopic coating of saliva materials. That covers the tooth surface
within seconds of eruption that is why it is exogenous origin (formed from substance outside the
tooth),,it is removed by polishing of the teeth

COMPOSITION : composed of structureless mass of primarily protein film that forms on erupted teeth

Involves selective adsorption of components from :

-Saliva

-Gingival crevicular fluid (GCF)

-Secretion from oral microbes

Chemical composition

•Glycoproteins

•Immunoglobulin

•Carbohydrates

glycoproteins include proline rich proteins that allow bacterial adhesion.

ROLE/ Significance of the acquired pellicle

Purpose of the acquired pellicle to cover the enamel of a tooth and protect it from acidic activity

(1)Protective: barrier against acids

(2) Lubrication: Keeps surfaces moist

(3) Nidus (a place in which bacteria have multiplied or may multiply; a focus of infection.)for bacteria

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(4) Calculus attachment

Types of pellicles

(1) Surface pellicle unstained: it's clear, translucent, insoluble, and is the initial film of glycoproteins

(2) Surface pellicle stained: it takes on extrinsic stain and becomes coloured because coffee, tea, or
smoking

(3) Subsurface pellicle: It's embedded in the tooth structure especially where demineralization occurs

2- Bacterial colonization:
The bacteria from the normal oral flora comes in contact with the pellicle ad adhere itself to it

The first to adhere are primary colonizers, sometimes referred to as "pioneer species” the primary
colonizers are not pathogenic. Since they are members of oral flora the primary colonizers are not
pathogenic. They are streptococci ,aerobic

But since the pellicle can be removed at this initial stage the both bacteria and pellicle can be removed
by mastication of foods, by the tongue, and by toothbrushing and other oral hygiene activities.

for this reason, bacteria tend to accumulate on teeth in sheltered, undisturbed environments, like
occlusal fissures, the surfaces apical to the contact between adjacent teeth, and in the gingival sulcus.

(i) Colonization by pioneer bacteria

Pioneer bacteria /early colonizers is the 1st group of bacteria to colonise.

Since they are members of oral flora the primary colonizers are not pathogenic. They have the
ability to adhere to tooth surface through the pellicle

The early colonisers / pioneer bacteria are :

* Streptococcus oralis

*Streptococcus mitis

*Streptococcus sanguinis

(ii) Bacterial Outgrowth

(iii) Bacterial Secondary Colonization

Those that arrive later are secondary colonizers. They may be able to colonize an existing bacterial
layer, but they are unable to act as primary colonizers.

(iv) Climax Community

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3-Plaque Matrix Formation

Initial plaque formation may take as long as 2 hours.

The bacteria begin to multiply

About 2 days are required for the plaque to double in mass, and the bacterial colonies are still growing
this happens and is accerlated by not using oral hygiene measures such as brushing and in areas where
is isolated away rom the masticatory forces

In case of regular brushing this step does not happen and is limited to areas which can not be reached
by the oral hygiene measures such as brushing and flossing

In 4-5 days the bacterial growth is the highest

Bacteria: short rods and filamentous bacteria. streptococci, the gram-positive facultative rods, and the
actinomycetes are the main organisms in plaque found in early fissures and proximal plaque.

As plaque oxygen levels fall, the proportions of gram-negative rods (e.g., fusobacteria) and gram-
negative cocci such as Veillonella increase

What bacteria do is:

Sucrose is used to synthesize intracellular polysaccharides that serve as an internal source of energy, as
well as external polysaccharide coats

THIN CARIOGENIC PLAQUE

•Plaque is THIN & POROUS.

•Is permeable to O2, saliva, GCF.

•Composed almost entirely of salivary protein. OPENED-STRUCTURE PLAQUE

Absence of Carbohydrates

Matrix : •Thin •Porous

•Permeable to : O2 Saliva GCF

THICK HEALTHY PLAQUE

•The formation of extracellular polysaccharides make the plaque matrix THICK & NON-POROUS.

•Composed of salivary protein & extracellular polysaccharides. CLOSED-STRUCTURE PLAQUE

Presence of Carbohydrates (Ex. Sucrose)

Matrix : Thick Nonporous

NON-Permeable to : O2 Saliva GCF

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4- Maturation of Plaque
plaque will turn into calculus after around 2 weeks

the deposition of calcium and other non-organic ions on the dental plaque is gradual and it gets harder
and more calcified as time goes by

at this stage the calculus can not be removed by regular oral hygiene measures and need the dentist
intervention

At times, the plaque demonstrates staining, which is caused by chromogenic bacteria, which produce a
brown pigment especially in smokers and person who consume colored substance in food and drinks
such as coffee ,tea and cola

Plaque control
1- Prevent or reduce the adhesion of early and/or subsequent bacterial colonisers
2- Prevent or inhibit the growth & proliferation of microorganisms
3- Prevent or inhibit the formation of the plaque matrix
4- Modify the biochemistry of plaque to reduce the formation of cytotoxic products
5- Modify plaque ecology to less pathogenic flora 6- Immunisation

How :

1- Personal mechanical practices/oral hygiene measures

Tooth brushing

•Flossing

Waxed thread

Minted thread

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Polishing

Stain removal

Regular polishing should be avoided at disease-free area

•Scaling & root planning

Directed at areas with clinical signs of disease

2- Surface Active Molecules

•Detergent action restrain bacterial colonization Antiseptics (bisbiguanide) eg. Chlorhexidine

3- Antimicrobial Control
1.Antimicrobial agents

2.Antiplaque agents

4- Immunisation
Vaccination using intact S. Mutans

Development of antibody which will result in Host defense against caries

Calculus
Calculus, or tartar, is the term used to describe calcified dental plaque.

•Calculus is mineralized plaque.

•Over 80% of adults have calculus, and its prevalence increases with age.

Calculus is a contributing factor to dental diseases.

•Calculus provides a rough surface on which bacterial plaque can grow

•Is classified according to its relation to the gingival margin

Supragingival Calculus

Mineralised plaque above the gingival margin

•Is visible in the oral cavity

•Minerals come from saliva

•Usually white in color

•Found mainly near salivary ducts

Subgingival Calculus

Mineralised plaque below the gingival margin. usually in the periodontal pockets.

•Is visible in the oral cavity

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Microbiology Oral Biology Dr. Radwa Khalil

•Minerals come from GCF

•Usually black in color

•Found everywhere

Calculus Composition
Composition of supragingival & subgingival calculus is different.

•Subgingival calculus has higher content of Ca and Mg.

Organic Component

•Protein polysaccharide complexes

•Epithelial cells

•Leukocytes

•Microorganisms

In-organic Component

•Consist of 70-90% inorganic salts

•Mainly in the form of :

Ca3 (PO4)2

Mg3(PO4)2

Rate of calculus formation

•Some people form calculus faster compare to other.

•The following factors increase the rate of calculus formation :

1.Elevated of salivary PH.

2.Elevated of salivary calcium concentration

3.Elevated of bacterial protein and lipid concentration

4.Low individual inhibitory factors

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Periodontal diseases
The accumulation of plaque and calculus especially in the gingival cervical sulcus will trigger the host
immune response to produce inflammation which is called

Gingivitis ;if the inflammation is restricted to the gingival area

Peritonitis :the inflammation spread to the periodontal tissue and alveolar bone

Healthy gingiva:

pink or coral pink color, (normal variations in color depend on race and complexion)

firm, resilient tissues

'orange-peel' texture(known as stippling)

shape that follows the contour of the teeth and forms a scalloped edge

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Microbiology Oral Biology Dr. Radwa Khalil

no areas of redness, swelling or inflammation

no bleeding during daily plaque removal

no discomfort

In case of gingivitis gums may be sore, bleed easily and appear puffy, soft and swollen.

No bone structure is lost around the teeth at this stage of gum disease.

Blood on the toothbrush or dental floss is one of the earliest and most common signs of gingivitis.

Your gums should never bleed while brushing or flossing.

gingivitis is that it is preventable and reversible through good brushing and flossing techniques (or other
interdental tooth cleansing).

On the other hand, if oral hygiene habits are poor, gingivitis may progress to periodontitis

periodontitis is irreversible

sings of periodontal disease:

gums that bleed when you brush or floss your teeth

gums that are red, swollen or tender

gums that have pulled away from teeth

infection including purulence (pus) between the teeth and gums when the gums are pressed

permanent teeth that are loose or separating

any changes in the way your teeth fit together when you bite

any changes in the fit of your partial denture

bad breath

itchy sensation

At the early stages of periodontitis :

Periodontitis occurs when the inflammation of the gums progresses into the deeper underlying
structures and bone.

In the most common form of periodontitis, plaque (and sometimes calculus) is found below the gum
line.

The gums may feel irritated, appear bright red, and bleed easily.

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The ligaments holding the tooth in its socket break down and the gums pull away from the teeth,
resulting in a periodontal pocket or space between the tooth and gum.

The periodontal pocket deepens and fills with more bacteria. Supportive ligaments and bone start to
show damage

Advanced periodontitis :

When periodontitis progresses to the advanced stage, the gums severely recede (pull away from the
tooth ); pockets deepen and may be filled with pus.

There may be swelling around the root and you may experience sensitivity to hot or cold or feel pain
when brushing your teeth. This is due to the severely receding gums exposing the root surface.

As bone loss increases, your teeth may lose so much support that they need to be removed to preserve
the overall health of your mouth

Treatment of periodontal disese :

Prevention

Detection

Assessment

Scaling, debridement, root surface planing

Periodontal surgery

Bone grafting

Regular dental visits

Dietary consideration

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Microbiology Oral Biology Dr. Radwa Khalil

Prevention Oral hygiene measures ,regular tooth brushing

and flossing
Dietary consideration
Detection Using disclosing agents
Signs and Symptoms
Bleeding, inflamed gums
Halitosis
Discoloration/Stain
Long in the tooth
Mobility
Radiographic findings
Periodontal probing depths
Assessment Regular dental visits
Scaling, debridement, root surface planning

Periodontal surgery In case of advanced peridontaitis

Bone grafting

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