PERIODONTAL

MICROBIOLOGY

Dr. Soni Bista

Department of Periodontology
Dr. SONI BISTA and
1Oral
st
yearImplantology
JUNIOR RESIDENT
PERIODONTOLOGY AND ORAL
PERIODONTIUM
 HUMAN MICROBIOTA

BIRTH CANAL:
STERILE Acquires vaginal & fecal
FETUS microorganism

>2yrs (after
Within 2 wks; weaning) ;
Newly mature Entire human
NEW
microbiota microbiota;
BORN
1014 microbial cell

Bacterial population in normal healthy human being: 2 kg of total body wt

 ORAL MICROBIOTA
Within hour after birth Facultative &
aerobic bacteria
1st & most
dominant
colonizer:
Streptococcus
salivaris &
Streptococcus
mitis
From 2nd day Anaerobic Bacteria

1st colonizers: Veillonela spp., Neisseria spp., Actinomyces

spp., Staphylococcus spp.
 COLONIZATION OF THE ORAL CAVITY

E Ko¨no¨nen, Oral colonization by anaerobic bacteria during childhood: role in health and disease.Oral Diseases
(1999) 5, 278–285
 COLONIZATION OF THE ORAL CAVITY

It is estimated that more

than 500 different species are
capable of colonizing the
adult mouth, and that any
individual may typically
harbor 150 or more different
species.

Eija Könönen . Development of oral bacterial flora in young children.Annals Of Medicine Vol. 32 , Iss. 2,2000
COMMENSAL MICROBIOTA

• Most oral bacteria are harmless commensals under normal

circumstances.
Specific conditions: D
1. Increased mass I
S
2. Increased pathogenicity
E
3. Suppression of A
commensal/beneficial bacteria S
4. Reduced host response E
ORAL CAVITY
FROM A
MICROBES
PERSPECTIVE
• Oral cavity is considered as an “open growth system” with
uninterrupted ingestion and removal of microorganisms and their
nutrients.

INTRAORAL SURFACES FOR BACTERIAL ADHESION

1. Intraoral, supragingival, hard surfaces (teeth, implants, restorations,
and prostheses)
2. Periodontal/periimplant pocket (with its crevicular fluid, the root
cementum or implant surface, and the pocket epithelium)
3. Buccal epithelium, palatal epithelium, and epithelium of the floor of
the mouth
4. Dorsum of the tongue
5. Tonsils
6. Saliva
 Non
Shedding
Shedding
surface
surface
NATURAL: Natural: Teeth,
Epithelial cells Nails
Intraoral epithelial cells

High turnover rate

Artificial:
Implants, Heart
Prevents accumulation of valves
microorganisms

Hard non shedding surfaces (teeth, implants):

1. Allows development of extensive structural bacterial deposit

2. Form a unique ectodermal interruption
 BIOLFILM

• Composed of microbial cells encased within a matrix of

extracellular polymeric substances (EPS) such as
polysachharides, proteins & nucleic acids

Heterogenous
Microcolonies
Water channels
Fronds
Steep chemical gradient
 DENTAL PLAQUE
• Defined clinically as structured, resilient, yello-greyish
substances that adhere tenaciously to the intraoral hard
surfaces, including removable & fixed restorations.
Bowen WH, 1976

Plaque is specific but highly variable structural entity

resulting from colonization of microorganism on tooth
surfaces, restoration and other parts of oral cavity which
consist of salivary component like mucin, desquamated
• Bacterial aggregations
epithelial on&the
cells, debris teeth or otherall
microorganism solid oral structures
embedded in a
Lindhe, 2003 WHO,1961
gelatinous extracellular matrix
 CLASSIFICATION

According to Listgarten
(1976)
1. Marginal plaque
2. Supragingival plaque
3. Subgingival plaque
 COMPOSITION
• OPEN FLUID FILLED CHANNELS

• MICROCOLONIES OF BACTERIA

• INTRACELLULAR MATRIX

INORGANIC:
ORGANIC: Calcium, phosphorus,
Polysachharides, sodium….
proteins, glycoproteins, 1. Supragingival: source is
DNA, albumin saliva
2. Subgingival: source is GCF
 SUPRAGINGIVAL PLAQUE

• Stratified organization of a multilayered accumulation of bacterial morphotypes

• Gram-positive cocci and short rods predominate at the tooth surface

• Gram-negative rods and filaments, spirochetes, predominate in the outer surface

of the mature plaque mass.
 SUBGINGIVAL PLAQUE

• Local availability of blood products

• reduction oxidation  anaerobic condition

• Gingival crevice/pocket is bathed by flow of GCF containing many substances that

bacteria may use as nutrients

• Host inflammatory cells and mediators are likely to have considerable influence
on the establishment and growth of bacteria in the subgingival region
 PLAQUE
FORMATION

1. FORMATION OF PELLICLE I. TRANSPORT TO THE

SURFACE
2. INITIAL ADHESION/ATTACHMENT OF BACTERIA
II. INITIAL ADHESION
3. COLONIZATION/ PLAQUE MATURATION
III. STRONG ATTACHMENT
 MICROBIAL COMPLEXES

Socransky et al. in 1998 examined over 13,000 subgingival

plaque samples from 185 adult subjects and DNA hybridization
methodology described the presence of specific complexes of
periodontal microorganisms.
 CRITERIA FOR IDENTIFICATION OF PERIODONTAL
PATHOGENS

In the 1870s, Robert Koch

postulated the criteria by
which an organism can be
judged to be causative agent
in human infections
Pathogen must be routinely isolated from the
diseased individuals.

Must be grown in pure culture in the laboratory.

Must produce a similar disease when inoculated into

susceptible lab animals.

Must be recovered from lesions in a diseased

laboratory animals.
 SIGMUND SOCRANSKY (1992) proposed criteria by which periodontal
microorganisms may be judged to be potential pathogens.

• Must be associated with disease, as evident by increases in the

1. number of organisms at diseased site.

• Must be eliminated or decreased in sites that demonstrate clinical

2. resolution of disease with treatment.

• Must demonstrate host response, in form of alteration in host cellular

3. or humoral immune reaction.

• Must be capable of causing disease in experimental animal models.

4.

• Must demonstrate virulence factors responsible for enabling the

5. microorganisms to cause destruction of the periodontal tissues.

A. Actinomycetemcomitans & P.gingivalis

 KEY CHARACTERISTICS OF SPECIFIC
PERIOPATHOGENS
1) AGGREGATIBACTER ACTINOMYCETEMCOMITANS

 Small, short (0.4-1 μm), straight or curved rod with rounded

ends.
 Nonmotile and gram negative.
 Multiple biotypes and five serotypes (a to e)
 Culture conditions and identification: Grows on TSBV agar
plate White, translucent colony with a star-shaped
internal structure
 1) AGGREGATIBACTOR
ACTINOMYCETEMCOMITANS

Special pathogenic characteristics:

• Possesses a number of virulence factors

1. Lipopolysaccharide

2. Leukotoxin

3. Collagenase

4. Protease
2) Tannerella forsythia
• Nonmotile, spindle-shaped, highly pleomorphic rod
• Gram-negative obligate anaerobe.

Culture conditions and identification:

1. Grows slowly (14 days for minute colonies)
2. Grows on blood agar plate: Smooth,
white colony with a faded edge.

Pathogenicity:
3. Produces several proteolytic enzymes that are able to destroy
immunoglobulins and factors of the complement system.
4. Induces apoptotic cell death.
3) Porphyromonas gingivalis
Special pathogenic characteristics:
1. Aggressivepleomorphic
• Nonmotile, periodontal pathogen.
(coccal to short) rod and gram-
2. Its fimbriae mediate adhesion, and its capsule defends
negative obligate anaerobe
against phagocytosis.
3. Produces a series of virulence factors, including many
proteases (e.g., for the destruction of immunoglobulins,
complement factors, and heme-sequestering proteins;
degradation of host cell collagenase inhibitors), a hemolysin,
and a collagenase.
Culture conditions and identification:
4. Inhibit migration of polymorphonuclear leukocytes (PMNs)
across
Darkanpigmentation
epithelial barrier and dark
(brown, affects the production
green, or blood
or black) on
degradation of cytokines by mammalian cells.
agar because of a metabolic end product from blood
5. Capacity to invade soft tissues.
(hemin).
4) Prevotella intermedia & Prevotella
nigrescens
• Prevotella group are short, round-ended, nonmotile,
gram-negative rods.
• P. intermedia and P. nigrescens are the most
pathogenic
Culture conditions and identification:
Grow anaerobically, with dark pigmentation (brown-
black colonies) on blood agar.

Special pathogenic characteristics:

• Less virulent and less proteolytic than P. gingivalis.
5) Campylobacter rectus
• Rare motile organisms involved in periodontitis.

• Gram-negative, short rod, curved (vibrio) or helical.

Culture conditions and identification:

• Grows anaerobically (anaerobic chamber or special

jars), with dark pigmentation when sulfide is added to
Special pathogenic characteristics:
thespecies,
This medium, which
as with is transformed to FeS,produces
A. actinomycetemcomitans, giving aa
gray stainC. rectus is less virulent and less proteolytic than P.
leukotoxin.
gingivalis.
6) Fusobacterium nucleatum
• Gram-negative, cigar-shaped bacillus with pointed ends
Culture conditions and identification:
Special pathogenic characteristics:
• Grows•Induce
in CVEapoptotic
agar plate: Round,
cell death flat rhizoid,and
in mononuclear opaque purple colony
polymorphonuclear cells
•Trigger the release of cytokines, elastase, and oxygen radicals
from leukocytes.

•Because fusobacteria coaggregate with most oral micro-

organisms, they are believed to be important bridging organisms
between the primary (early) and secondary (late) colonizers during
colonization.
7) Spirochetes
• Represent a diverse group of spiral, motile organisms
• Are helical rods 5-15 μm long with a diameter of 0.5
μm.
• Have three to eight irregular spirals. Their cell wall is
gram negative, but they stain poorly.

• Classified species of spirochetes include Treponema

denticola, Treponema vincentii, Treponema socranskii
(often associated with periodontitis), and Treponema
pallidum (associated with secondary syphilis).
7) Spirochetes
Culture conditions and identification:
• Extremely difficult to grow and need strict anaerobic
conditions and a specific medium.

Special pathogenic characteristics:

• Ability to travel trough viscous environments enables them
to migrate within the gingival crevicular fluid and to
penetrate both the epithelium and the connective tissue.
• Some spirochetes have the capacity to degrade collagen
and even dentin.
• T. denticola produces proteolytic enzymes that can destroy
immunoglobulins (IgA, IgM, IgG) or complement factors.