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• Introduction
• Taxonomy
• terminologies
• Classification
• Ecosystem
• Oral microflora
• Oral econiches
• Ecological relationships
• Microbial complexes
• Colonization of micro-organisms in the form of biofilm
• Specific bacteria in periodontal diseases
• Association of plaque microorganisms with
periodontal disease
• Conclusion
• References

• A microorganism (from the Greek: mikros, "small" and
organismós, "organism") is a microscopic organism,
which may be a single cell or a multi-cellular organism.
• Single-celled microorganisms were the first forms of
life to develop on Earth, approximately 3–4 billion
years ago.
• The possibility that microorganisms exist was
discussed for many centuries before their discovery in
the 17th century.

.TAXONOMY • Taxonomy • Classification of living organisms into groups • Phylogenetic Classification System: • Groups reflect genetic similarity and evolutionary relatedness • Phenetic Classification System: • Groups do not necessarily reflect genetic similarity or evolutionary relatedness. observable characteristics. • Instead. groups are based on convenient.

TERMINOLOGIES • Species: A collection of microbial strains that share many properties and differ significantly from other groups of strains • Strain: A culture derived from a single parent that differs in structure or metabolism from other cultures of that species .

prokaryotic variant strains characterized by biochemical or physiological differences Morphovars .Type strain: One of the first strain among the species.prokaryotic variant strains characterized by distinctive .prokaryotic variant strains characterized by morpholigical characteristics Serovars . studied and fully characterised than other strains. Biovars .

• Symbioses: both partners benefit • Commensalism : one partner benefits. other is neutral • Parasitism: one partner benefits while the other is harmed • Mutualism: different organisms living together .

Mechanism of pathogenesis Transmission Adherence Invasion Colonization Damage Exit Survival .


. They are almost always unicellular.Prokaryotes  Prokaryotes are organisms that lack a cell nucleus and the other membrane bound organelles.  Consisting of two domains. bacteria and archaea.

slime layer ...flagella 2.capsules 4.CELL STRUCTURE OF PROKARYOTE 1..pili (fimbriae) 3..

and can function and reproduce as individual cells. For example. ARCHAEA • Archaea differ from bacteria in both their genetics and biochemistry.BACTERIA • They lack a nucleus and other membranebound organelles. archaean membranes are made of ether lipids. Their genome is usually a single loop of DNA. . while bacterial cell membranes are made from phosphoglycerides with ester bonds. but often aggregate in multicellular colonies.

including humans. • Unlike bacteria and archaea. .• EUKARYOTES • Most living things that are visible to the naked eye in their adult form are eukaryotes. the Golgi apparatus and mitochondria in their cells. eukaryotes contain organelles such as the cell nucleus.







 Capnophiles. • ANAEROBIC : That does not require oxygen for growth.ATMOSPHERE • AEROBIC : That can survive and grow in an oxygenated environment.   A microaerophile is a microorganism that requires oxygen to survive. Eg:Campylobacter spp. but requires environments containing lower levels of oxygen than are present in the atmosphere. . require an elevated concentration of carbon dioxide to survive.

Bacteroides. which cannot use oxygen for growth. Streptococcus spp.. Prevotella.ANAEROBIC • Obligate anaerobes.. Listeria spp. Fusobacterium. Clostridium. . Staphylococcus spp. Escherichia coli. which can grow without oxygen but use oxygen if it is present. Porphyromonas. • Aerotolerant organisms. but tolerate its presence • Facultative anaerobes.. which are harmed by the presence of oxygen.

5 3 .  MESOPHILIC: temperatures between 20 -45 degrees.  THERMOPHILIC : temperatures above 60 degrees  BACTERIAL METABOLISM pH : • Neutrophil • Acidophil 6–8 • Alkalophil 10.TEMPERATURE  PSYCHROPHILIC: Temperatures below 15 degrees.







29 .


ECOLOGICAL NICHE & • A niche is a term describing the way of life of a species. • The ecological niche describes how an organism or population responds to the distribution of resources and competitors and how it in turn alters those same factors for its benefit.  The ecosystem is composed of microbial communities living on specific sites surrounded by a different physical and chemical elements. unique niche. ECOSYSTEM • Each species is thought to have a separate." .  Definitions of  Hutchinson: “The set of biotic and abiotic conditions in which a species is able to persist and maintain stable population sizes.

Ways of obtaining and using






• Oral microflora refers to the community of
microorganisms coexisting in the oral cavity as its
primary habitat;
• These strains of bacteria colonize the various
different surfaces present in the oral cavity, and
communicate between each other through
complex cell signaling processes;
• Over time as the individual is further exposed to
external sources of bacteria, the biodiversity of the
oral cavity increases, to a point where stability is
• This is termed the climax community.

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).
• 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.

• Established during infancy . • Colonization from source sites and caregiver saliva. . • Tooth eruption provides nonshedding surfaces: • The “window of infectivity” concept.BACTERIA DURING THE LIFE CYCLE • Oral colonization begins in the birth canal: • Populations on the tongue and mucosa.include anaerobes.

• Complete loss of teeth shifts flora towards infant state: • Dentures restore supragingival non-shedding sites.• Hormonal shifts . • Implants restore supra.puberty and pregnancy: • Can alter proportions of Gm.and subgingival sites.anerobes. .

ORAL FLORA CHANGES WITH AGE Time during a lifetime MAJOR COMPONENTS & CHANGES IN ORAL FLORA Newborn Oral cavity sterile. New habitats include hard surfaces and gingival crevice. Spirochaetes isolated more frequently . & Fusobacteria Tooth eruption Increase in complexity. Soon colonised by facultative and aerobic organisms. Veillonella sp. Child to adult Various anaerobes frequently found inc. S sanguis. esp S. salivarius 6 months Flora becomes more complex & includes anaerobic orgs eg. S mutans and A viscosus appear. Members of the Bacteroidaceae.

modify it and makes it convenient for resettlement of new microbial population. • Growing microbial community was enriched with different species living in complex.FORMATION OF THE ECOSYSTEM • The development of microbial community comprises an alternation of populations. • The process of alternation of populations and enrichment of the community stops only when niches for colonization of new microbial species are unavailable. • The process starts with the colonization of the environment of the first microbial population. • It fills the new space. .

means that a variety of sites are available for colonization by oral microorganisms. • Each site has unique characteristics and allows those microorganisms best suited to the environment to inhabit the site. tongue and mucosa. . such as teeth.DIVERSE ECOLOGICAL NICHES IN THE ORAL CAVITY • The heterogeneity of tissue types in the oral cavity.

Or Blowing The Nose (2) Tongue And Oral Hygiene Implements (3) The Wash-out Effect Of The Salivary. • Most organisms can only survive in the oropharynx when they adhere to either the soft tissues or the hard surfaces (teeth. Mastication. And Crevicular Fluid Outflow (4) Active Motion Of The Cilia (Nasal And Sinus Walls).• Dynamic Equilibrium: (1) Swallowing. dentures. Nasal. 40 . and implants).


palate. . Low biomass (reservoir) sites. but proportions may differ. High biomass sites: • Non-shedding surfaces: • Supragingival tooth surfaces. • Subgingival tooth surfaces. external gingiva. • Shedding oral mucosal surfaces: • Buccal.ORAL ECOLOGICAL ZONES • Mostly the same species are present. Saliva as a transitional zone. floor of mouth. • Shedding surface: • The tongue.

palate. lips. tongue. • Supragingival. • Soft structures – mucosa: • Cheeks. . • Keratinized and nonkeratinized mucosa. gingiva. providing various locations for colonization: • Subgingival. • Epithelium of the gingival sulcus.ORAL ECONICHES • Hard structures – teeth.

which have cells which are constantly replaced due to the normal wear-and-tear of the mouth. can be found on the dorsum of the tongue. • Hence species not found elsewhere.MUCOSAL SURFACES: • These include the palate. cheek and tongue. • The different mucosal surfaces also have different properties which contribute to the presence of different types of bacteria: • The presence of the numerous crypts on tongue allows for bacteria to be protected from the normal shedding and removal by saliva flow. such as obligate anaerobes. .

• Can live and grow inside. . • Can direct the cell to export them to other cells. • Bacteria “subvert” the cell to take them in: • Take control of the cytoskeleton.MUCOSAL RESERVOIR SITES • Some oral species can invade epithelial cells: • This requires communication between bacteria and cells. • Multi-species intracellular flora resembles mixed biofilm.

• Veillonella. S.. • Gram . • Obligate anaerobes black . • Gram + rods .. S.MUCOSA OF THE GINGIVA. . • S. • Peptostreptococcus spp. FLOOR OF THE MOUTH Surface of the tongue • Streptococcus: • Streptococcus salivarius. CHEEK. mitis.Bacteroides spp. • Veillonella spp. • Haemophilus. oralis.rods an spirochetes associated with periodontal diseases..Actinomyces spp. sanguis. PALATE.rods . • Neisseria.

• Gram positive MO;
• Facultative anaerobes:
• Streptococcus;
• Actinomyces;

• Gram negative:
• Veillonella, Haemophilus,

 From

healthy gingival sulcus
 Gram

negative rods:














• They are the narrow crevice between gingival
epithelium and cementum
• They have low oxygen tension which is
favorable for Gm- anaerobes;
• Major site for interaction between bacteria
and host tissues;
• Species mix varies
between each side
and the center which
form distinct

• Antimicrobial factors. • They are the basis for maintaining oral ecosystem: • Provide water.ORAL FLUIDS • Oral surfaces washes of two liquids: • Saliva. • Nutrients. • Gingival fluid. • Adhesion. .

. • Most of the microbes present in the mouth utilize the glycoproteins and proteins in the saliva as their main source of nutrition. ensuring the optimal growth of the resident colonies. • Proteins and glycoproteins of saliva are responsible for the formation of the pellicle. saliva maintains the pH of the mouth. • By acting as a buffer.MICROORGANISMS IN SALIVA • Microbial composition of saliva is the most similar to the tongue.

the main source of microbial transmission between individuals. • Saliva is carrying bacteria .SALIVARY FLOW • It is responsible for the removal of nonendogenous bacteria which is unable to adhere to specific sites in the mouth. • Areas which receive markedly less saliva flow. . • Saliva also circulates bacteria within the oral cavity. resulting in re-colonization of oral surfaces where the microflora might be removed via mechanical forces such as cleaning. proximal spaces and occlusal fissures. tend to have significantly higher levels of bacterial buildup. such as deep gingival crevices.

• The flow of this fluid removes foreign microbes
which do not adhere to these surfaces;
• For the resident population, the flow of
crevicular fluid provides these organisms with a
source of nutrition.

• In a healthy body, the internal tissues
- blood, brain, muscle, etc., are
normally free of microorganisms;
• However, the surface tissues - skin
and mucous membranes, are
constantly in contact with
environmental organisms and become
readily colonized by various microbial
• species;
The mixture of organisms regularly found at any
anatomical site is referred to as the normal
flora or resident flora, some researchers
prefer the term "indigenous microbiota".

• Typical microflora of a econiche;
• Microorganisms are separated and grouped according to
the different conditions of life;
• Resident microflora has an important function in host:
• Digestive and nutritional;
• Competition with pathogenic microflora.

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

The presence of friendly resident microorganisms on oral
surfaces contributes to the body’s defense against
foreign pathogens, which are generally transient and
can give rise to harmful infections.

This is known as colonization resistance;

which inhibits infections via competition for resources. • Essential nutrients derived from the saliva and various proteins in the oral cavity are also more effectively utilized by the resident microflora. . which reduces the available sites left for the attachment of exogenous organisms.FUNCTION OF MICROFLORA • Colonization resistance is firstly achieved through the saturation of oral surfaces with preexisting resident bacteria.

and they contribute to immunity by inducing low levels of circulating and secretory antibodies that may cross react with pathogens. peroxides and bacteriocins. • Contribute to host nutrition through the synthesis of vitamins. .THE NORMAL BACTERIAL FLORA OF THE ORAL CAVITY • Benefit from the host who provides nutrients and habitat. • Exert microbial antagonism against exogenous species by production of inhibitory substances such as fatty acids. • Occupy available colonization sites which makes it more difficult for other microorganisms to become established.

.TRANSIENT MICROBIOTA • Transient microbes are just passing through. transients are unable to remain in the body for extended periods of time due to: • Competition from resident microbes. • Although they may attempt to colonize the same areas of the body as do resident microbiota. if the opportunity arises. • Physical or chemical changes within the body that discourage the growth of transient microbes.  However. resident and transient microbes cause the host no harm. some of these microbes are able to cause disease and become opportunistic pathogens. • Elimination by the body’s immune system. OPPORTUNISTIC MICROBES  Under normal conditions.

THIS CAN HAPPEN DUE TO A NUMBER OF DIFFERENT CONDITIONS: • When the immune system is compromised or suppressed. normal flora can overpopulate or move into areas of the body where they do not normally occur. microbes that are normally crowded out by resident microbes have an opportunity to take over. . for example when a person takes broad spectrum antibiotics. • Disease can result when normal flora are traumatically introduced to an area of the body that they do not normally occur in. • When the balance of normal microbes is disrupted.

but has previously been inapparent or dormant. as well as the atmosphere.ENDOGENOUS MICROFLORA •  That is microflora already present in the body. •  Exogenous bacteria can be either be benign or pathogenic. . •  They exist in aquatic and terrestrial environments. EXOGENOUS MICROFLORA • Exogenous bacteria are microorganisms introduced to closed biological systems from the external world.

.  Synergistic. management or control of other organisms.ECOLOGICAL RELATIONSHIPS • Independence • Life free from influences.  Antagonistic microbial interactions. • Dependence MICROBIAL RELATIONSHIPS  There are a complex of relationships between different microbial species:  Neutral.

gingivalis and F. .NEUTRAL  Relationship between two species living together without affecting each other . SYNERGY • Relationships between the two microbial species living together. wherein both are mutually favored.  P.neither favorable nor unfavorable. Example of synergy  Various microorganisms may cooperate to use substances which they are not able to metabolise themselves. leading to enhancement of the effects of each of these. Nucleatum both can hydrolyze casein.

Mechanisms of microbial  Competition for adhesion receptors or for nutrients.  Hydrogen peroxide:  Dairy complex.  Bacteriocins.ANTAGONISM • Relationship of two microbial species living together. .  Antibiotics.  Enzymes. and less effect on the independent operation of each one of them. yielding inhibition of their function. antagonism  Production of inhibitory substances:  Organic fatty acids.

• Separation by bacteriocins inhibit gram positive microorganisms.sanguis and S. . and gram negative microorganisms.AN EXAMPLE OF MICROBIAL ANTAGONISM • S. • It inhibits the growth of S. mutans and Lactobacillus produce lactic acid thereby producing an acidic environment. oralis.

• Neglected oral hygiene: • Anaerobes. • Oral hygiene and antimicrobial agents : • Good oral hygiene: • Facultative aerobes. • Proteolytic microorganisms. • A diet high in protein inhibits the development of Lactobacillus. • Acidogenious microorganisms. .INFLUENCE OF EXTERNAL FACTORS • Diets : Food high in carbohydrates increases the development of Str. mutans and Lactobacillus. • Drugs and diseases.

reduced buffering capacity.ANTIMICROBIAL AGENTS • Fluorides have depressing effect on microbial attachment. • Suppress the acid tolerance of gram + microorganisms. . • Reducing sugar transport. • Reduce their glycolytic activity.  Antibiotics suppress the resident microflora. DRUGS AND DISEASES  Patients with reduced salivation have reduced capacity to remove sugars. leading to over-development of antibiotic resistant species Candida and allow colonization of exogenous pathogens such as Enterobacteriaceae.

ADVERSE EFFECTS • Source of endogenous infection. • Deliver certain biological factors to the host. • Change the local environment to facilitate exogenous microbial overdevelopment.BENEFICIAL EFFECT OF THE MICROFLORA ON THE BODY • Prevent exogenous and endogenous infections. . • Stimulate an immune response. • Create hypersensitivity in organism to microbial agents. • Help in renewal and healing of the epithelium.

Mitis Strep. aureus Streptococcus mutans Strep. Salivarius Strep. sanguis Strep. Faecalis Beta-hemolytic streptococci GRAM-NEGATIVE FACULTATIVE RODS Enterobacteriaceae Hemophilus influenzae Eikenella corrodens Actinobacillus Actinomycetemcomitans .HUMAN ORAL FLORA GRAM-POSITIVE FACULTATIVE COCCI Staphylococcus epidermidis Staph.

Flavescens . Viscosus Lactobacillus GRAM-NEGATIVE AEROBIC OR FACULTATIVE COCCI Eubacterium Neisseria sicca N. odonotolyticus A.GRAM-POSITIVE ANAEROBIC COCCI Peptostreptococcus sp GRAM-NEGATIVE ANAEROBIC COCCI Diphtheroids Corynebacterium GRAM-POSITIVE ANAEROBIC RODS Actinomyces israelii A.

Protozoa Mycoplasma Entamoeba gingivalis Tirchomonas tenax Mycoplasma orale M.SPIROCHETES YEASTS Treponema denticola T. pneumoniae . Microdentium Candida albicans Geotrichum sp.

produce lactic acid.IMPORTANT ORAL BACTERIA 1. role in dentine caries rather than enamel caries. • Streptococcus is facultative anaerobic cocci which produces lactic acid Some are implicated in caries. • Cell wall has thick peptidoglycan layer (penicillin has effect by interfering production of this layer). . • Most are fermentative. THREE IMPORTANT GENERA: • Actinomyces is facultative anaerobe. • Lactobacillus. facultative anaerobe. cocci or irregular shape (pleomorphic). Gram Positive organisms: • Rods (bacilli). • Oxygen tolerance varies from aerobes to strict anaerobes.

Extracellular Polysaccharide Production of Acid Cariogeni Endoc carditis isolates S mutans + + + +++ + S sanguis + + + ++ ++ S mitis + _ + _ + + _ +++ S milleri + _ + + + S salivarius _ _ + _ _ .IMPORTANCE OF STREPTOCOCCI IN THE ORAL AND THEIR PROPERTIES Importan t Oral Species Growth on hard surfaces Production of Insol.

salivarius _ ++ ++ _ . mitis +++ +++ +++ +++ S.DISTRIBUTION OF STREPTOCOCCI IN THE ORAL CAVITY Species Cheek Tongue Saliva Tooth S.mutans _ _ +/- ++ S.

rods. • Range of oxygen tolerance but most important strict or facultative anaerobes. others produce enzymes which break down tissue.2. especially in established/subgingival plaque. • Mainly consist of Cocci. filamantous rods. . produce acids which other organisms use acids as an energy source. GRAM NEGATIVE ORGANISMS • Many Gram-negative bacteria are found in the mouth. spindle shaped or spiral shaped. • Some fermentative.

actinomycetemcomitans associated with aggressive periodontitis. . • Neisseria. intermedia • Fusobacterium: F. • Actinobacillus/Aggregatibacter: A.e ANUG. nucleatum periodontal pathogen.MOST IMPORTANT GRAM NEGATIVE BACTERIA: • Porphyromonas: P. • Treponema: group important in acute periodontal conditions i. gingivalis • Prevotella: P. • Veillonella.

FLORA OF NORMAL. fungi. HEALTHY DENTATE MOUTH % (approx) 85% Remainder Bacteria Streptococci Veillonella Gram positive Diptheroids Gram negative anaerobic rods 5-7% Neissaeria 2% Lactobacilli 1% Staphylococci & Micrococci Other bacteria. protozoa & viruses .

tropicalis. parapsilosis. C.angular chelitis  Medium used – sabouraand’s agar(peptoneglucose)  Indentification – psuedohyphae. C.erythematous candidiasis. C.hyperplastic candiasis.FUNGI IN THE ORAL CAVITY  Most commonly found :. C.albicans. septate hyphae 77 .candida species (C. guilliermondi)  Other rhodotorula & torulopsis ( denture wearer)  Conditions – thrush . stellatoidea.

periodontal pockets and infection of tonsils  Can become opportunistic pathogen  T.PARASITES IN THE ORAL CAITY ENTAMOEBA GINGIVALIS TRICHOMONAS TENAX E.tenax –  only parasitic flagellate in  oral cavity  --number increases in periodontitis 78 . gingivalis –  found in soft calculus.

4. pH. Temperature. 7. 6. Host genetics (changes in immune response etc). . 2. Nutrients (endogenous & exogenous (diet). REDOX Potential / Anaerobiosis. 3. 5. Host Defences (Innate & Acquired immunity). Antimicrobial agents & inhibitors.FACTORS AFFECTING GROWTH OF MICROORGANISMS IN THE ORAL CAVITY 1.

5 – 7. • Str. mutans.9 .7 and 7. OR HYDROGEN ION CONCENTRATION • In the mouth pH varies between 6. . In the gingival fluid pH is 7. • When microorganisms colonize they are exposed to extreme temperatures. • The temperature is variable on the teeth and mucosa.TEMPERATURE • Relatively constant .5 .5.34˚-36 ˚. In the gingival sulcus pH is alkaline -7.3. • • • In an acid medium in the eco niches following are developed: • Lactobacillus.8. PH. • It is maintained by saliva through salivary flow and buffer systems.

• Positive redox potential are seen in buccal and palatal mucosa and the posterior part of the tongue. • In a predominance. . • These processes depend on the oxygen and their redox potential. reduction processes have a negative redox potential and develop anaerobic microorganisms. and others to reduction.REDOX POTENTIAL AND • Under the action of the enzymes some of the ANAEROBIOSIS components are subjected to oxidation. • Negative redox potential are seen in approximal surfaces. fissures and gingival sulcus.

• Products of metabolism of other microbial species. •  Genetic factors. • Residues from host`s food. • Saliva. HOST FACTORS • Host defense mechanisms. • Hormonal changes.NUTRIENTS • Desquamated epithelial cells. •  Stress. • Gingival fluid. .

sensors (Toll-like receptors). • Salivary antimicrobial factors • Mucosal antibodies (secretory IgA). • Innate immunity . In most cases. host defenses tolerate oral bacteria • The predominant relationships are commensal. • Cell-mediated immunity (T-cells).RELATIONSHIPS WITH THE HOST Host defenses in the mouth: • Epithelial cells: • Barrier function. • Inflammatory mediators. antimicrobial peptides. .

. • Lactoferrin. • Antimicrobial factors: • Lysozyme. • Complement.HOST DEFENSE MECHANISMS • Remove the microorganisms through stimulation of salivary flow. • Leukocytes. • Histatine-rich peptides. • Nonspecific protection: • Mucin. • Specific protection: • Salivary Ig A. • Cistatin. • Salivary peroxidase.

.REMOVAL OF THE • The majority of microorganisms in the mouth is removed MICROORGANISMS by washing action of saliva. bacteria can adhere to host cells and form a commensal relationship. In essence. • Salivary flow is stimulated by muscle activity of lips and tongue. • One of the most important mechanisms is shedding. especially of the gingiva. this is a natural cleansing mechanism. • The soft tissue surfaces employ a variety of mechanisms to prevent adhesion of pathogenic organisms. • However. • The high turnover rate of the intraoral epithelial cells. beneficial for both parties. prevents the permanent accumulation of large masses of microorganisms on these surfaces.

.SECRETORY IGA SYSTEM • SIgA-antibodies reduce microbial adhesion to enamel epithelium and through: • Neutralizing enzymes • Neutralize toxins • Synergy with other antibacterial agents such as lysozyme. • Protects the mucosa of penetration of antigens. • Helps complement activation.SPECIFIC PROTECTION . peroxidase and mucin. lactoferrin.

• As part of pelicle protects teeth from demineralization. • Catches microorganisms and antigens like in a trap. . • Limits microorganisms penetration into tissues. • Eliminates microorganisms with continuous updating of mucin layer combined with washing action of saliva flow.MUCIN • Provides a protective coating of enamel and mucosa.


Socransky et al (1998) MICROBIAL ADHESINS • Constructed of: • Polysaccharides. rather there are specific associations among bacterial species.ADHESION Adhesins can be of the microbial surface and receptors on oral surfaces The association of bacteria within mixed biofilms is not random. Fibrils. • Lipoteichoic acid. • Glycosyltransferase. . Capsules. • Carbohydrate-binding proteins. • They are located in the cell wall in the form of: Fimbriae.

• Amylase. • Glucans. • Proline-rich proteins.RECEPTORS ON THE ORAL STRUCTURES SALIVARY COMPONENTS BACTERIAL COMPONENTS • Mucin. • Lysozyme. • Glycoproteins. IgG. • IgA. . • Glycosyltransferase.

. BACTERIAL ATTACHMENT TO A SURFACE CAN BE DIVIDED INTO SEVERAL DISTINCT PHASES • Primary and reversible adhesion. • Biofilm formation. • Secondary and irreversible adhesion.

• In both cases.CO-AGGREGATION • Coaggregation interactions are believed to contribute to the development of biofilms by two routes: • The first route is by single cells in suspension specifically recognizing and adhering to genetically distinct cells in the developing biofilm. • The second route is by the prior coaggregation in suspension of secondary colonizers followed by the subsequent adhesion of this coaggregate to the developing biofilm. . bacterial cells in suspension (planktonic cells) specifically adhere to cells in the biofilm in a process known as coadhesion.

• Strengthens bacterial attachment. • A dose: There is a certain amount of microorganisms. • Frequency of exposure: Partial colonization. . ADHESION IS DEPENDENT ON: • A contact: Start of interaction.ECOLOGICAL SIGNIFICANCE OF BACTERIAL COAGGREGATION • Specific coaggregation processes are likely to have an important ecological role as an integral process in the development and maintenance of mixed-species biofilm communities. • With specific receptors on the cell surfaces. • Adsorption: • By electrostatic forces to the surface of pellicle. • Increases the stability of the plaque matrix.

• Bacterial cells in biofilms can also produce elastases and cellulases which become concentrated in the local matrix and produce tissue damage.• Bacterial cells within biofilms can produce enzymes such as beta-lactamase against antibiotics. catalases. and superoxide dismutases against oxidizing ions released by phagocytes. • These enzymes are released into the matrix producing an almost impregnable line of defense. 94 .

• Lower plaque layer are dense in which microbes are bound together in polysaccharide matrix with other organic and imorganic materials. • On top of lower layer. • Biofilms can facilitate processing. cross feeding.DENTAL BIOFILM • Biofilm is a highly organized structure and consists of microcolonies of bacterial cells randomly distributed in a shaped matrix or glycocalyx. removal of harmful metabolites and development of appropriate physicochemical environment. uptake of nutrients. • Microcolonies occur in different shapes according to shear forces. loose layer can be seen which can extend into surrounding medium (for teeth and saliva) • The fluid layer bordering the biofilm has a stationary sublayer and a fluid layer in motion • Nutrient components penetrate this fluid medium by molecular diffusion • Biofilms act as primitive circulatory system. 95 .

mass transfer. it has open fluid filled channels running across the plaque mass • Plaque is a naturally-constructed biofilm. nutrient concentrations and quorum sensing influence biofilm properties . • Changes in shear forces. which result in dental disease. in which the consortia of bacteria may reach a thickness of 300-500 cells on the surfaces of the teeth. • These accumulations subject the teeth and gingival tissues to high concentrations of bacterial metabolites. • High rate of reproduction and physiological adaptation to environmental resources help biofilms to survive and sustain.BIOFILM • The classic biofilm that involves components of the normal flora of the oral cavity is the formation of dental plaque on the teeth.


• In addition. salts. proteins. are the major components of the biofilm making up 50–95% of the dry weight. produced by the bacteria in the biofilm.EXOPOLYSACCHARIDES – THE BACKBONE OF BIOFILMS • The bulk of the biofilm consists of the matrix or glycocalyx and is composed predominantly of water and aqueous solutes. • They play a major role in maintaining the integrity of the biofilm as well as preventing desiccation and attack by harmful agents. they may also bind essential nutrients such as cations to create a local nutritionally rich 98 . • Exopolysaccharides (EPS). • The “dry” material is a mixture of Exopolysaccharides. and cell material.

• They can exist in ordered and disordered forms. • The EPS matrix could also act as a buffer and assist in the retention of extracellular enzymes (and their substrates) enhancing substrate utilization by bacterial cells.• They can be neutral or charged polyanionic macromolecules and different concentrations can alter the conformation of the gel network. • The density of fibrillar masses and the flexibility and configuration changes could affect accessibility to nutrients and solutes. .

and when it’s time to go. • Different genes are active in planktonic and biofilm states. . • Bacteria at the outer surface of mature biofilms are signaled to detach and become planktonic.• Quorum sensing tells bacteria when to grow.

F. 2° colonizers (Gram-) • Bridge species .1° colonizers (Gram+) • Streptococci bind pellicle proteins from saliva DENT 5302. nucleatum • Bind other bacteria 3° colonizers (Gram-) • Porphyromonas gingivalis ECOLOGICAL SUCCESSION .

• Excrete lactic acid. • Starch-digesting enzyme. .INTER-SPECIES COMMUNICATION • Streptococci ferment CHO. • Strep can make amylase. • Enhances lactate excretion. • They are biofilm buddies. • Veillonella use lactate made by Strep for nutrition.


• Binding to strep improves survival when O2 is present. .INTERSPECIES COLLABORATION O 2 Streptococcus cristatus: • Facultative species: Fusobacterium nucleatum: • Robust anaerobe. • Coaggregation essential to survival when O2 is present. Porphyromonas gingivalis: • Sensitive anaerobe.

• Streptococcus cristatus thus coaggregates with F. • S.COLLABORATIVE INVASION • F. cristatus does not invade cells. nucleatum • Fusobacterium nucleatum is considered a bridge species because it is a promiscuous coaggregator. nucleatum. S. nucleatum invades epithelial cells. . cristatus is carried inside by F. • After coaggregation.

Actinomyces naeslundii 9. Streptococcus mitis 4. Streptococcus gordonii 2.Primary Colonizer s 1. Streptococcus intermedius 3. Streptococcus sanguinis 6.Capnocytophaga sputigena 14.Aggregatibacter actinomycetemcomitans serotype a 11.Veillonella parvula 106 .Actinomyces odontolyticus 16.Capnocytophaga ochracea 13. Actinomyces israelii 8.Capnocytophaga gingivalis 12. Actinomyces oris 10. Streptococcus oralis 5. Actinomyces gerencseria 7.Eik enella corrodens 15.

15. 13. 11. 9. 2. 5. 10. 16. 8.Secondar y Colonizer s 1. 6. 4. 7. 12. 3. 14. 17. Campylobacter gracilis Campylobacter rectus Campylobacter showae Eubacterium nodatum Aggregatibacter actinomycetemcomitans serotype b Fusobacterium nucleatum ssp nucleatum Fusobacterium nucleatum ssp vincentii Fusobacterium nucleatum ssp polymorphum Fusobacterium periodonticum Parvimonas micra Prevotella intermedia Prevotella loescheii Prevotella nigrescens Streptococcus constellatus Tannerella forsythia Porphyromonas gingivalis Treponema denticola 107 .

Nonbacterial microorganisms that are found in plaque include Mycoplasma species.  Deep pocket . One gram of plaque (wet weight) contains approximately 10 11 bacteria.  Healthy crevice - 103 bacteria. In a periodontal pocket. yeasts.108 bacteria. protozoa.STRUCTURE AND COMPOSISTION OF DENTAL PLAQUE1 Dental plaque is composed primarily of microorganisms. 108 . and viruses.

gingivalis. intermedia T. St. P. F. Forsythis. intermedius Peptostreptcoccus micros P.Subgingival Tissue associated St. oralis. Nucleatum 109 .

some anaerobic Mainly anaerobic Energy source Mainly ferment carbohydrate Many proteolytic forms Motility Few Many Pathology Caries & gingivitis Gingivitis & periodontitis . branching rods. filaments & spirochaetes Mainly rods & spirochaetes Energy Metabolism Facultative.DIFFERENCE BETWEEN MATURE SUPRA & SUB-GINGIVAL PLAQUE Characteristic Supra gingival Sub gingival Grams stain Gram + or -ve Mainly Gram –ve Morphotypes Cocci.

Most important group in periodontal pathgenesis .• Attached plaque: tooth associated and predominantly composed of gram +ve rods and cocci • Cause subgingival calculus and root caries • Unattached plaque: extends to frontier of apical plaque and have gram –ve organisms. Form the bioactive area and cause large amount of inflammatory GCF • Epithelium associated: loosely attached to gingival epithelium and contain gram –ve and motile rods.

112 .

MICROBIOLOGY OF PERIODONTAL DISEASES ARE THERE TRUE ORAL PATHOGENS? • Classic concept of a pathogen • Not normally present • Produces “virulence factors” • Damage host directly (e.g. toxins) • Induce host to damage itself (immune responses) • Presumed oral pathogens don’t quite fit that model • Normally present throughout life • Damage requires presence in large numbers • Ecological concept of oral microbial diseases • Ecological shifts lead to changes in proportions • Balance shifts in favor of “pathogens”/disease .

• This phenomenon is consistent with other infectious diseases in which it may be observed that a pathogen is necessary but not sufficient for a disease to occur. 114 . • In spite of the presence of periodontal pathogens.SIMILARITY BETWEEN PERIODONTAL DISEASES AND OTHER INFECTIOUS DISEASES • Individuals may be colonized continuously by periodontal pathogens at or below the gingival margin and yet not show evidence of ongoing or previous periodontal destruction. periodontal tissue damage does not take place.

if exposed. to the epithelial surfaces of the gingiva or periodontal pocket.• UNIQUE FEATURES OF PERIODONTAL The major reason forINFECTION this uniqueness is the unusual anatomic feature that a mineralized structure. • Thus. a situation is set up in which microorganisms colonize a relatively stable surface. and to other bacteria which are attached to these surfaces. • The tooth provides a surface for the colonization of a diverse array of bacterial species. so that part of it is exposed to the external environment while part is within the connective tissues. to underlying connective tissues. • Bacteria may attach to the tooth itself. the tooth and are continually held in immediate proximity to the soft tissues of the periodontium. the tooth passes through the integument. 115 .

• The organisms that cause periodontal diseases reside in biofilms that exist on tooth or epithelial surfaces. spirochetes. amoeba. HISTORICAL PERSPECTIVE • Investigators in the period from 1880–1930 suggested four distinct groups of microorganisms as possible etiologic agents. fusiforms. and streptococci. • The biofilm provides a protective environment for the colonizing organisms and fosters metabolic properties that would not be possible if the species existed in a free-living (planktonic) state. 116 .


• AMOEBA They found higher proportions of amoeba in lesions of destructive periodontal diseases than in samples taken from sites in healthy mouths or mouths with gingivitis. 118 . • The role of amoeba in periodontal disease was questioned by some authors because amoeba were found in sites with minimal or no disease and could not be detected in many sites with destructive disease and because of the failure of emitin to ameliorate the symptoms of the disease. SPIROCHETES • Investigators reported higher proportions of spirochetes and other motile forms in lesions of destructive disease when compared with control sites in the same or other individuals.

STREPTOCOCCI • These microorganisms were proposed on the basis of cultural examination of samples of plaque from subgingival sites of periodontal disease. 119 . • Vincent (1899) distinguished certain pseudomembranous lesions of the oral cavity and throat from diphtheria and recognized the important role of fusiforms and spirochetes in this disease. • These organisms were originally recognized on the basis of their frequent appearance in microscopic examination of subgingival plaque samples. were the spindle-shaped fusiforms.•FUSIFORMS The third group of organisms that were frequently suggested to be etiologic agents of destructive periodontal diseases. • The selection of the streptococci may have been predicated upon the fact these were the only species that could be consistently isolated from periodontitis lesions using the cultural techniques of that era. • The organisms were first related to periodontal disease by Plaut (1894). including Vincent’s infection.

non-motile. capnophilic. Slots 1982). This is a small. (Newman et al 1976. actinomycetemcomitans with the 530 bp deletion were 22.5 times more likely to convert to LJP (Bueno et al 1998). 120 . Subjects harboring A.ACTINOBACILLUS ACTINOMYCETEMCOMITANS Aclinobaciltus" refers to the internal star-shaped morphology of its bacterial colonies on solid media (Colebrook 1920) and to the short rod or bacillary shape of individual cells (Lieske 1921. round-ended rod. Gram-negative. saccharolytic. Chung et al 1989) . Slots 1980. This species was first recognized as a possible periodontal pathogen in lesions of localized juvenile periodontitis.

Recently. Haemophilus aphrophilus and Haemophilus segnis to a new genus Aggregatibacter gen.It was originally named Bacterium actinomyeetum comilans (Klinger 1912) which was changed to Bacterium comitans (Lieske 1921) and finally to Actinobacilius actinomycetemcomitans (Topley & Wilson 1929). nov. nov. Actinobacillus actinomycetemcomitans. The species of the genus Aggregatibacter are independent of X factor and variably dependent on V factor for growth in vitro. Norskov-Lauritsen N. Kilian M (2007) 121 . as Aggregatibacter actinomycetemcomitans comb.

122 .

anaerobic. Burdon 1928). P.PORPHYROMONAS GINGIVALIS: It is a Gram negative. non motile. melaninogenicus (Bacterium melaninogenicum. gingivalis is a member of the much investigated “black pigmented Bacteroides” group. Initially they were grouped into a single species. 123 . asaccharolytic rods that usually exhibit coccal to short rod morphologies. B.

forsythus. highly pleomorphic rod. spindle shaped. B. 1979) as a “fusiform” Bacteroides. was first described in 1979 (Tanner et al. B. forsythus was detected more frequently and in higher numbers in active periodontal lesions than inactive lesions (Dzink et al 1988). 124 . anaerobic. This species has been shown to produce trypsin like proteolytic activity (BANA test positive. Loesche et al 1992) and induce apoptotic cell death (Arakawa et al 2000).TANNERELLA FORSYTHIA Third consensus periodontal pathogen. The organism is a Gram negative.

SPIROCHETES These are Gram negative.  Clearly.  The organism has been considered as possible periodontal pathogens since the late 1800s and in the 1980s. 125 . highly motile microorganisms that are common in many periodontal pockets. anaerobic. At least 15 species of subgingival spirochetes have been described.  Difficulty in distinguishing individual species. a spirochete has been implicated as the likely etiologic agent of acute necrotizing ulcerative gingivitis (Listgarten & Socransky 1964). helical shaped.

Yuan et al 2001). These “were the most frequently detected spirochetes in supra and subgingival plaques of periodontitis patients. Pathogen related oral spirochetes” (PROS) were also shown to have the ability to penetrate a tissue barrier in in vitro systems (Riviere et al 1991). denticola was found to be more common in periodontally diseased than healthy sites (Haffajee et al 1998.T. 126 .

Papapanou et al 2000). 127 . round-ended anaerobic rod. It is a Gram negative. and also in certain forms of periodontitis (Herrera et al 2000. short. intermedia is the second black pigmented Bacteroides to receive considerable interest in pathogenesis of chronic periodontitis. They have been shown to be particularly elevated in acute necrotizing ulcerative gingivitis (Loesche et al 1982).PREVOTELLA INTERMEDIA/PREVOTELLA NIGRESCENS P.

It has also been shown to invade oral epithelial cells in vitro (Dorn et al 1998). intermedia that show identical phenotypic traits have been separated into two species. nigrescens (Shah & Gharbia 1992). gingivalis and was shown to induce mixed infections. P. (Hafstrom & Dahlen 1997). Strains of P. 128 . intermedia and P.This species appears to have a number of virulence properties exhibited by P.

a small rod: thus. Acts as “microbial bridge” facilitating coaggregation 129 between early and Late colonizers. . The species can induce apoptotic cell death in mononuclear and polymorphonuclear cells (Jewett et al 2001). and animalis.. The name Fusobacterium has its origin in fusus. which belongs to the family Bacteroidaceae.FUSOBACTERIUM NUCLEATUM F. fusiforme. Gharbia and Shah (1990) divided Fusobacterium species into four subspecies: subspecies nucleatum. (Moore et al 1985). polymorphum. nucleatum is the type species of the genus Fusobacterium. and bacterion. a small spindle-shaped rod. This species is the most common isolate found in cultural studies of subgingival plaque samples comprising app. 7-10% of total isolates. a spindle.

C. 130 . anaerobic. It was first described as a member of the “vibrio corroders”. The organism is unusual in that it utilizes H2 or formate as its energy source. short. motile vibrio. These are the only two oral species known to possess this characteristic (Gillespie et al 1992). actinomycetemcomitans. rectus has been shown to produce a leukotoxin. rectus is a Gram negative.(Rams et al 1993). eventually shown to include members of a new genus Wolinella and Eikenella corrodens.CAMPYLOBACTER RECTUS C. Like A. It was found in higher numbers in disease sites as compared with healthy sites and more in sites exhibiting active periodontal destruction.

corrodens has been shown to stimulate the production of matrix metalloproteinase (Dahan et al 2001) and IL-6 and IL-8 (Yumoto et al 1999).EIKENELLA CORRODENS E. capnophilic. asacharolytic. small rod with blunt ends. In tissue culture system. E. This species was found more frequently in sites of periodontal destruction as compared with healthy sites and higher levels in active sites (Tanner et al 1987). regular. corrodens is a Gram negative. 131 .

anaerobic. small. 132 . 2000). P. Riggio et al 2001). micros is a Gram positive. Two genotypes can be distinguished with the smooth genotype being more frequently associated with periodontitis lesions than the rough genotype (Kremer et al. micros was found to be in higher numbers at sites of periodontal destruction as compared with healthy sites (Papapanou et al 2000.PEPTOSTREPTOCOCCUS MICROS P. asaccharolytic coccus.

tumbling motility and. 133 . are Gram negative.SELENOMONAS SPECIES The selenomonas spp. saccharolytic rods and may be recognized by their curved shape. curved. by the presence of a tuft of flagella inserted in the concave side. Moore et al (1987) described six genetically and phenotypically distinct groups isolated from oral cavity and found at a higher proportion of shallow sites (PD>4mm) in chronic periodontitis. in good preparations.

strictly anaerobic. nodatum. Eubacterium brachy and Eubacterium timidum are Gram positive. E. Some of these species elicited elevated antibody responses in subjects with destructive periodontitis. small somewhat pleomorphic rods. (Moore et al 1985).EUBACTERIUM SPECIES Certain Eubacterium species have been suggested as possible periodontal pathogens due to their increased levels in disease sites. (Martin et al 1988) 134 .


with approximately 104 to 106 bacteria.GINGIVITIS  The microbial load at diseased sites is greater. Equal proportions of gram positive (56%) and gram negative (44%) species. which uses the steroid as growth factors 136 .  The microbiota of chronic gingivitis (plaque induced) consist of app.  Pregnancy associated gingivitis is accompanied by increases in steroid hormones in crevicular fluid and dramatic increases in levels of P. intermedia. as well as facultative (59%) and anaerobic (41%) microorganisms.

P. Certain gram-negative bacteria with pronounced virulence properties have been strongly implicated as etiologic agents (e.CHRONIC PERIODONTITIS Cultivation of plaque microorganisms from sites of chronic periodontitis reveals high percentages of anaerobic (90%) and gram negative (75%) bacterial species. 137 .g. gingivalis and Tannerella forsythensis).

the most numerous isolates are several species from the genera Eubacterium. naeslundii. and in still others P. Actinobacillus actinomycetemcomitans plays a causative role in LAP. gingivalis may be etiologically more important. nucleatum. A. especially in cases in which patients harbor highly leukotoxic strains of the organism.LOCALIZED AGGRESSIVE PERIODONTITIS The microflora of subgingival biofilms from patients with LAP is similar to that of patients with chronic periodontitis and is predominantly composed of gram negative. capnophilic. and anaerobic rods. However. F. some populations of patients with LAP do not harbor A. and Veillonella parvula. rectus. C. actinomycetemcomitans. 138 . On a percentage basis.

gingivalis.GENERALIZED AGGRESSIVE PERIODONTITIS The subgingival flora in patients with generalized aggressive periodontitis resembles that in other forms of periodontitis. actinomycetemcomitans. and Campylobacter species. T. The predominant subgingival bacteria in patients with generalized aggressive periodontitis are P. forsythensis A. 139 .

gingivalis and F. nucleatum accounting for 7-8% and 3.4%. but it is clear that they constitute a very large and diverse group. 140 .NECROTIZING ULCERATIVE GINGIVITIS/PERIODONTITIS The majority of the spirochetes (treponemes) associated with necrotizing ulcerative gingivitis are uncultivable. More than 50% of the isolated species were strict anaerobes with P. respectively.

and Candida albicans have also been detected. gingivalis (50. P.1%). P. and T. intermedia (62. coagulase-negative staphylococci. 141 . nucleatum (70.5%). forsythensis (47. An average of approximately 70% of the cultivable flora in exudates from periodontal abscesses are gram-negative and about 50% are anaerobic rods. micros (70. Enteric bacteria. P.6%).0%).PERIODONTAL ABSCESSES The bacteria isolated from abscesses are similar to those associated with chronic and aggressive forms of periodontitis.8%). Periodontal abscesses revealed a high prevalence of the following putative pathogens: F.

Other species such as Pseudomonas aeruginosa. forsythia. are also frequently detected around implants. 142 . micros. P. Species such as A. Candida albicans and staphylococci. and Capnocytophaga are often isolated from failing sites. gingivalis. actinomycetemcomitans.PERIIMPLANTITIS Studies have shown that microbiota associated with periimplantitis are comparable to that of periodontitis (high proportion of anaerobic gram negative rods. Fusobacterium. and spirochetes). enterobacteriaceae. rectus. motile organisms. P. C. T.

• Plaque samples were removed from the diseased site. • These vaccines were administered systemically or locally in the periodontal tissues. 143 . autogenous vaccines. and other oral organisms. These included vaccines prepared from pure cultures of streptococci. either in the local periodontal lesion or systemically. and/or by immersion in iodine or formalin solutions. while others using the same techniques were more sceptical.VACCINES • Three types of vaccines were employed for the control of periodontal diseases. • Autogenous vaccines were prepared from the dental plaque of patients with destructive periodontal diseases. then re-injected into the same patient. and stock vaccines such as Van Cott’s vaccine. • Proponents of all three techniques claimed great efficacy for the vaccination methods employed. “sterilized” by heat. Goldenberg’s vaccine or Inava Endocorps vaccine.

SPECIFIC BACTERIAL BEHAVIOUR IN BIOFILM: ANTIBIOTIC RESISTANCE • Microorganisms in biofilm are 1000 to 1500 times more resistant to antibiotics than in their planktonic stage • The mechanism of this increased resistance differs from species to species. from antibiotic to antibiotic. and for biofilm growing in different habitats RESISTANCE OF BACTERIA TO ANTIBIOTICS IS AFFECTED BY THEIR • Nutritional status • Growth rate • Temparature • pH • Prior exposure to subeffective concentration of anti microbial agents 144 .

• Also slower growth of bacterial species in biofilm is another important mechanism of antibiotic resistance • Biofilm matrix although not significant barrier in itself to diffusion of antibiotics but have certain properties to resist diffusion • • Biofilm act as ion-exchange resin removing Some antibiotics such as Macrolide which are antibiotics from solution positive charged but hydrophobic are unaffected by this process 145 .

resistant pump that can extrude antimicrobials from the cell 146 .• Also extracellular enzymes such as β lactamases. formaldehyde lyase and formaldehyde dehydrogenase may become trapped and concentrated in the extracellular matrix thus inactivating some antibiotics(especially positive charged hydrophilic antibiotics) • “Super-resistant” bacteria have been identified within a biofilm and these cells have multidrug.

147 .

Full mouth disinfection has been introduced by Leuven group in the 1990s • This strategy attempts to eradicate. and saliva) 148 . but also from all their intraoral habitats(mucous membrane. one stage. or atleast suppress.TRANSLOCATION AND MECHANICAL DEBRIDEMENT • To reduce chance of intraoral transmission. periopathogens in short time not only from the periodontal pockets. tongue.

Periodontology 2000. Vol. 78-1 6. 1994. Microbial etiology of Periodontitis. Blackwell munksgaard. Periodontology 2000. Haffajee & sigmund. 9th edition 3. Textbook of Microbiology. 16–22 5. 4th edition.REFERENCES 1. 1). . Microbial etiological agents of destructive periodontal diseases. 4. Anne. Clinical Periodontology and Implant dentistry. D. Jan Lindhe. 14–26.S. PD Marsh: Plaque as a biofilm: pharmacological principles of drug delivery and action in the sub. 5. Socransky.and supragingival environment. Carranza’s clinical periodontology. Vol. 2004.Anantnarayan 2. Oral Diseases (2003) 9 (Suppl. Tatsuj Nishihara & Takeyoshi koseki. 36.