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DEVELOPMENT OF ORAL MICROFLORA

Dr PRATHIBHA RANI .S Post Graduate Student

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INTRODUCTION

CONTENTS
Introduction History Prokaryotes and Eukaryotes Bacteria -Cell structure -Bacterial physiology -Bacterial taxonomy and classification Microbes of oral environment Origins Development of oral flora
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CONTENTS
Adhesion of oral flora Growth of oral flora Survival of oral flora Distribution of oral flora Gram positive, gram negative species of oral cavity Fungi and viruses of oral cavity Micro flora of Dental caries, Periodontal disease & Endodontic infections

 History of Microbiology In 1660s Robert Hook Microbiology began in the mouth: Anton-van Leeuwenhoek developed and used the first microscope to examine material collected from teeth, and described motile animalcules.

After gap of two centuries-Pasteur and koch Robert Koch first to prove that the micro organisms caused disease His postulates Specific M.O Should be isolated and grown Should produce disease when inoculated in healthy individuals Re-isolated in pure culture
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COMPARISION OF PROKARYOTES & EUKARYOTES

CHARACTERISTIC Nuclear membrane Cell wall Ribosomes Endoplasmic reticulum Golgi complex Lysosomes Actin filaments Peroxisomes Nucleolus Mitochondria Chromosomes Cilia/flagella Microtubules Prokaryotes Yes Yes Yes No No No No No No No Single No No Eukaryotes Yes No Yes Yes Yes Yes yes yes Yes Yes Multiple Yes
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Yes

Bacterial Structure And Physiology

Shape and size -spherical or rod shaped, clusters/chains Gram stain -Gram positive-Thick amorphous cell wall, retains fixed violet dye. -Gram negative-Layered appearance, stain is washed out.

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 Surface appendages( motility & adhesion) Flagella Fimbriae The cytoplasm Sporulation Bacterial physiology -Nutrition -Energy production
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 Growth of bacteria -Definition -Binary fission

Bacterial growth curve -Lag phase -Logarthmic/Exponential phase -Stationary phase

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Bacterial Taxonomy And Classification

Classification Identification Nomenclature

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THE NORMAL FLORA AND PATHOGENS OF HUMAN BODY

Commensal flora Pathogens Bacterial survival in extremes Thermophilic->45 deg c. Psychrophilic-<20 deg. Halophilic-Salt env. Acidophilic-Acid env. Basophilic-High ammonia.
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ANTIMICROBIAL FACTORS IN HEALTH

Mechanical ;Flushing action of liquids, Peristalsis of gut, skin, cough/sneeze, mucus, shedding of epithelial cells, cilia. Bio-chemical; Anaerobicity/ acidity, sebaceous secretions, sweat,lysozyme,antimicrobial enzyme,digestive enzymes,bile, detergent action,colonisation resistance. Immunological; complement,phagocytosis,inflammation,acute phase response,antibodies,cell mediated responses.
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ORAL CAVITY
The attainment of symbiotic status b/w some organisms with their host has occurred over the long period of evolutionary change and these organisms are referred to as Autochthonous biota Normant,latent,carrier state &masking &unmasking of viruses.

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The Oral Flora In 1979, Bowden & collegues isolated about 21 genera comprising 60 sp. Can be recognized as three types Indigenous floraStreptococcus,Actinomyces & Neisseria
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Supplemental flora;
Eg.Lactobacillus spp found in low levels in plaque, reduced plaque ph- become dominant.

Transient flora;
- Lack of mechanisms to persist Quickly disappear

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Host bacteria inter-relationship

Can be demonstrated in one of the three ways-

Symbiosis-Eg;digestive tracts of ruminants & termites,subsist on diet in large proportion of cellulose, gut bacteria secreting cellulase reduce cellulose to residues absorbed by the host
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Anti-biosis;

Amphibiosis ; E.g.; growth of S .mutans & B. gingivalis do not compromise the survival of the host
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Growth promoting Warmth,moisture, nutrients

Growth limiting Antibacterial,limited nutrients,exfoliation,swall owing.

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ORIGINS OF ORAL MICROBIOTA

In utero During birth - Corynebacterium, lactobacilli, coliformis,micrococci,anaerobic strepto, anaerobic cocci, protozoa, yeasts and some viruses. Oral cavity considered sterile until first breath is taken. Pioneer species - St.salivarius Others staphylo, lactobacilli strepto, pneumo, entero, coliforms sarcinae ,hemophili, Candida albicans.

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Established pioneer community By the end of third month Identifiable resident micro-flora At the end of one year sp. of strep, veilonella, staphylococci, neisseria identified

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 Major change at around 6 months Organisms- hard enamel surfaces & dento gingival margin S.mutans & S.sanguis- enamel surfaces. Climax community Around 300 species

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At Birth- Sterile, simple,aerobic Strept,Staphy,Colifor ms, Pioneer sppStrep.salivarius Climax communityStaph.albus,N.spp,Vei lonella spp,Candida.albicans
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 Infancy & Early childhoodS.mitis,S.sanguis,S.mutans,Ne isseria,Actinomyces,Lactobacil li,Rothia.Fusobacterium,Veilon ella. Adolescence-Deep fissures,Large interproximal areas,Deep gingival crevice Climax communityHard tissuesStrep.mutans,Strep.sanguis,A ctinomyces G CPrevotella,Porphyromonas,Ba cteroides,Spirochetes,Leptotric hia,Fusobacterium,Vibrio
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 AdulthoodPeriodontal diseaseSpirochetes,Bacteroides.

PlaqueStrep,S.mutans,S.mitior,S .sanguis,Actinomyces,Gra m+ve,Gram-ve filaments

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 Edentulous adultsSecond childhood-loss S.sanguis,S.mutans,Anae robes,Few Bacteroides,Spirochetes, More Yeasts Denture plaquecomposition similar to plaque on the tooth surface or at the junction of the tooth and the gingiva.
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ORAL FLORAL DEVELOPMENT

Adhesion- cell-substratum
adhesion,homotypic cell to cell & heterotypic cell-cell First phase - Initial reaction -Deposition -Extra cellular carbohydrate influence -Teeth and saliva Homotypic cell adhesion Heterotypic cell adhesion

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Adhesion factors
A range of physicochemical positive and negative forces along with surface features-shape, chemical composition & charge Weak attachment-Strengthen by polymer bridges Fine filaments
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Role of host polymers

Saliva-suspending medium composed of lipids,vitamins,ions,,polysaccharid es & immunoglobulin Acquired pellicle- Epithelial surfaces & teeth, composed of glycoprotein & antibodies.
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Role of bacterial polymers

No. of org.produce extra cellular polysaccharides to adhere to hard surfaces S.mutans- glucosyl transferase and fructosyl transferase Type of polymers with these interactions first- water insoluble - mutan second - water soluble dextran third - unusal soluble fructan Lipoteichoic acid bacterial polymer - wall of +ve organism
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Site specific receptors (adhesins)

Fuzzy coat/ Glycocalyx -St.mitior ,S.salivarius Trypsin sensitive, Polysaccharide & lipoteichoic acid. Fimbriae (pili) Sex pilus - Mating b/w cells Somatic pilus Adhesion Lectins-Receptors on tooth surface.

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Physical Retention
Organisms that lack good adhesive properties - Veilonella, Bacteroids & Spirochetes take refuge in dental plaque

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Growth of oral flora

Temperature- Psychrophils;0-30 dg C -Mesophils;10-45 dg C -Thermophils-25-75 dg Temp. affects-Ph,ion activity, solubility of gases & aggregation of macromolecules Oral cavity-stable heaven

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 Acidity; -Most org require ph close to neutrality -Saliva regulates ph -Ph of plaque falls following consumption of sugar to values - 5 or 4 -Aciduric -Acidogenic -Saliva - neutralising effect -Depth of penetration of saliva into dental plaque is doubtful
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 Oxidation reduction potential (Eh);

1. 2. 3. 4. Obligate anaerobes Facultative bacteria Obligate aerobes Micro aerophilic OxidationReduction potential(Eh) Represented by positive and negative symbol Healthy gingival crevice +73mv Disease - 300mv
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 Carbon di oxide; needed for growth initiation & continued growth Atmospheric co2 is 0.03% Many bacteria require increased levels of co2 E.g. - Actinobacillus & capnocytophaga linked with priodontal diseases. Nutrients; Can be obtained from saliva & GCF Saliva - Provide org sub AA, proteins, sugars, glycoprotein which can promote the growth Conversely no. of components may inhibit the growth E.g. Lysosomes, peroxidases & IgA
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Nutrients;

GCF contains no. of factors ,IgA ,IgG ,IgM & albumin Hemin & alpha-2 globulin for Bacteroides & Treponema denticola resp. GC harbors a lot of enzymes. Primary feeders Secondary feeders

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SURVIVAL OF THE FLORA

Depends on the ability to withstand the defensive system of the body. Some inhibit phagocyte engulfment while others prevent phagocyte digestion. Can be related to bacterial capsule Polysaccharides surface components such as Hyaluronic acid. Intracellular pathogens- Offer no resistance but after ingestion exist in balance with the phagocyte. Extra cellular pathogens- Destroyed as soon as engulfed.
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 Strepto pyogenes evolved anti phagocytic capsule Bacteroids- factors that inhibit migration & phagocytosis of PMNS. catalase, super oxide dismutase Strepto& Actino Leukocidin Capnocytophaga -changes in PMNS morpho.& impair complement induced PMNS chemo taxis. Bacteroides-non-specific fibrolysin
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ORAL FLORA DISTRIBUTION

Depends on certain factors Salivary secretions, GCF, microbial nutrition & metabolism, oral hygiene practices, systemic diseases ,oral habits ,diet,Eh, microbial interactions ,oral & dental diseases, racial & genetic factors , hormonal secretions, anatomical factors, drugs & anti microbial agents , microbial adhesions, dental treatment.

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ORAL FLORA ENVIRONMENTAL FACTORS

Two specific surfaces, Hard teeth & soft -epithelial surfaces Tongue - specialized surface

1)Lips Transitional zone Skin flora Staphylo.aureus , micro, gram positive rods, Oral flora-gram negative spp.also,Neisseria & Veilonella

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 Cheeks- Three predominant spp strept. mitior, strepto. sanguis, S.salivarius Palate -Similar to that of cheeks, no of strepto ,lacto,& hemophilus Tongue- Due to presence of crypts and papillae-greater bacterial density than elsewhere S.salivarius(50%), S.mitior, S.milleri & S.sanguis,Hemophili, Neisseria, Veilonella, Lactobacilli, Bacteroides, Fusobacterium to certain extent S.mucilagenosus - throat & nasopharynx produce polysaccharide slime.
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Saliva - Mucin an organic content favours the aggregation & adherence of bacteria such S.sanguis & S.mitis Carbohydrate do not influence the bacterial growth 20 AA ,Urea by filtration from blood Ammonia derived from urea-Provide source of nitrogenous sub for bacterial growth Teeth- S.mutans, S.sanguis, S.mitior, S.milleri,Actinomyces.
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 Gingival crevice 1. Differs considerably from saliva 2. Harbors a community of M O 3. Flushing action provides a protective action ( as in periodontal diseases ) 4. Proteins such as albumin, immunoglobulin IgG IgA & IgM as well as complement ,monocyte, lymphocytes of T & B type. 5. As depth increases the Eh & mixture of organisms gets altered. 6. S.mitior, S.sanguis ,S.salivarius ,Entero cocci, gram +ve org, Veilonella & Neisseria,
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MEMBERS OF ORAL FLORA

Gram positive Gram negative Fungi Viruses

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Gram positive Facultative anaerobic cocciG.Enterococcus,Stomatococcus,Streptococcus Obligate anaerobic cocciG.Peptostrepto Regular non-sporulating & facultative anaerob.rodG.Lactobacillus Irregular,non,facultatively anaerobic rodsG.Actinomyces,G.Arachnia,G.Bacterionema,G.Rothia Irregular,non,obligate,anaero,rodsG.Bifidobacterium,G.Eubacterium,G.Propionibacteriu m
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Gram negative Facultative anaerobic cocciG.Neisseria, Obligate anaerobic cocci-G.veilonella F.anaerobic,rodsG.Actinomyces,g.Capnocytophaga,G.Eikenella, G.Hemophilus Obligate anaerobic,straight,curved,helical rodsG.Bacteroides,G.Fusobacterium,G.Leptotrichia,G. Selenomonas,G.Wolinella Spirochetes-G.Treponema Fungi- G.Candida Viruses- Herpes V
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Gram positive bacteria

Streptococci; pairs/chains, non sporulating, nonmotile, most numerous single group(25-50%) Occur in oral cavity, upper res. tract
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Mutans Group Of Streptrococci

Species S. mutans S. sobrinus S. rattus S. cricetus dg b ab Serogroup cef + + + Mannitol Sorbitol + + + +
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STREPTOCOCCUS MUTANS
First discovered Clarke in 1924 Primary habitat Tooth surface of man Gram positive cocci which occur in pairs or as short or medium length chains-mutans refer to ability to form long chains when grown in sucrose medium. S.mutans carious lesions Can be divided into 8 serotypes a to h Supported on cell wall analysis, poly acrylamide gel, electrophoresis of proteins ,DNA content ,DNA hybridization studies Can synthesise soluble & insoluble extra cellular polymer from sucroses (glucan, mutan,fructan)

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Streptococcus sanguis
Pri. habitat dental plaque Colonization-after eruption of deciduous teeth Can be isolated from blood & heart valves in SBE Spherical, oval cells occur in medium or long chains Extracelluar soluble and insoluble glucans from sucrose Properties common S.mutans Not an instigator of D.C
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Asso. with Aphthous stomatitis.

Streptococcus mitior
Group of alpha hemolytic strep. Spherical or oval cocci found singly/pairs/chains Alpha or beta hemolysis on blood agar Extra cellular polysaccharide very little Found to be adherent to non keratinized cells (tongue, cheek & lips) Role in DC not proved
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Streptococcus milleri
S. angionosus Spherical,/ovoid cells in pairs/chains, long/short chains Hemolysis alpha ,beta, gamma Do not produce polymers from glucose Normal in habitat isolated from plaque, gingival crevice & throat. Cariogenic in gnotobiotic animals Isolated from infections abscesses of mouth, brain, liver appendix & blood stream
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Streptococcus rattus
Gram positive coccus, pair/chains Fermentation occurs with mannitol, sorbitol, raffinose, sucrose, lactose, maltose not with glycerol or xylose Extra cellular glucan from sucrose Less common in humans

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Streptococcus salivarius
Spherical/ovoid cells variable chain length Majority are non hemolytic on blood agar some alpha and beta hemolytic Do not produce sticky dextrans -thus does not colonize plaque/tooth surfaces. One of the first organisms to colonize the infants mouth In habitat-tongue fissures, Isolated from blood-Infective endocarditis No caries inducing properties Regarded as the True commensal

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Streptococcus cricetus
Cocci are gram positive pairs/chains Some produce alpha hemolysis majority are non hemolytic Best grown in reduced atmosphere Fermentation Mannitol sorbitol, rafinose, mannose, sucrose& lactose Originating from serovar type a Can not produce ammonia from arginine Isolated occasionally in man
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Streptococcus sobrinus
Gram positive in pairs/chains of varying length Some alpha hemolysis others non-hemolytic Fermentation of mannitol, insulin &lactose Ammonia is not produced from arginine Habitat- tooth surface induce DC in gnotobiots

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Streptococcus ferus
Gram positive in pairs/chains Can produce extra cellular & intracellular glucans Ferment- mannitol & sorbitol No fermentation of rafinose Not isolated from human mouth

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Streptococcus mitis
Appear spherical/ellipsoidal form long chains Ferment-glucose, sucrose, maltose, lactose No fermentation of mannitol, inulin, glycerol or xylose Isolated from human saliva, respiratory tract & feces. Serologically-similar to S.sanguis DNA hybridization show links with S.mitior

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Actinomyces
Are filamentous bacteria Obligate or facultative anaerobic, capnophilic, grm positive & non acid fast A.viscosus , A.naesludi & A.odontolyticus have limited patogenecity A .israeli & A. bovis implicated in actinomycosis of man & lumpy jaw in cattles
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 A. viscosus - facultative anaerobic, grow in presence of CO2,produces pdl disease, isolated from dental calculus, related to root surface caries A. naeslundii - common in habitat of oral cavity, tonsillar crypts, plaque & calculus, produce pdl disease A. odontolyticus- located carious teeth on blood agar produce green area similar to hemolytic strep

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 A .israeli- normal member of oral flora, tonsillar crypts & dental calculus Actinomycosis a true opportunistic infection invading tissues & regions of body as portals of entry occur A bovis- lumpy jaw of cattles not transmitted to humans
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1.

Propionibacteria;
Anaerobic gram positive rods which produce a high peak of propionic acid in glucose broth.

2.

Isolated from dental plaque& oral cavity.Primary habitat-skin.

3.

P.acnes isolated from oral cavity in association with dental plaque,carious dentine & necrotic pulp tissue.

4.

Association DC,pdl diseases not proven

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 Bacterionema B .matruchotii; identified in oral cavity gram positive, on acid fast, on motile, facultative Cells - pleomorphic whip-handle cells Ferment carbohydrates to acetic, propionic & lactic acid acts as an important focus for the calcification of plaque

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Lactobacilli
Currently 43 species of this genus have been listed long, slender, bent rods chain formation observed most are non motile& non sporulating gram positive, later become gram negative -age They are both acidogenic and aciduric Homo fermentative -lactic acid Hetero fermentative when less of lactic acid is produced. In oral cavity L. acidophilus & L. salivarius homo fermentors break down glucose via emb meyerhof pathway & high lactic acid production
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 Other two sp L .fermentum, L .breve's Hetero fermentors degrading glucose by a 6- phospho gluconate pathway producing lactic acid,ethanol or acetic acid Other two species L. casei & L .plantarum use both pathways called facultative hetero fermentors.

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GRAM NEGATIVE BACTERIA Neisseria-Gram negative aerobic/ facultatively anaerobic cocci ,isolated from the tongue, lips & cheek, plaque & saliva. N. sicca & N. catarrhalis in oral cavity. Infection of oral mucous membrane

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 Veilonella-strictly anaerobic cocci comprise 5-10% from saliva & tongue surfaces. V. parvula &V .alcaseus. Lack glucokinase & fructokinase..Use pyruvate, lactate, maltate, fumarate or oxaloacetate for their growth Cause few infections & Is protective to the oral cavity.
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 Hemophili - aerobic/ fac. anaerobic

gram negative rods , isolated from oral cavity. Enter oral cavity during infancy but have low toxicity. Opportunistic cause endogenous infections, otitis media & infective endocarditis H .segnis common mem of dental plaque.
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Eikenella corrodens - Due to its property to pit/corrode the surface of agar plates . Small, non sporulating, non encapsulated, non motile micro aerophilic, gram negative. Isolated from oral cavity, abscesses of various parts of the body.
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 Bacteroids -30 species, occur in large bowel & faces. In oral cavity divided into pigmented & non-pigmented. B. melaninogenicus is further subdivided isolated from periodontal pockets.
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 Fusobacteria Anaerobic, sporulating, gm ve rods, with fusiform shape. paired rods giving elongated cigar app. first noted in ulcerative gingivitis in 1880,later linked to the presence of spirochetes in vincets infection. F. nucleatum -oral cavity its increase associ.with periodontitis
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 Wolinella - gm ve curved anaerobic rods with single polar flagellum. W.succinogenes-abdominal inf. Dental abscess . W .recta in gingival crevice. Capnocytophaga-gm ve fusiform rods, capnophilic, Possible role in periodontitis & juvenile periodontitis

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FUNGI
Candida only fungal spp accepted as members of the oral commensal flora Candidiasis local/systemic Yeast like fungus In oral cavity no. of sp isolatedC.albicans, C.tropicalis, C .krusei, C .parapsilosis, C guilliermondii & C. glabrata C .albicans-Acute/Chronic
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candidiasis

 Acute-pseudo membranous ,atrophic candidiasis Chronic-Atrophic ,hyper plastic, muco cutaneous Acute pseudo membranous seen in young childrenconsist of dead mucosal cells with fungal hyphae. Acute atropic Imbalance of normal flora due to use of antibiotics

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 Chronic atropic swollen inflamed mucosa in denture wearers linked with angular chelitis Chronic hyperplastic- firm white, persistent plaques on the lips, tongue & cheeks Oral candidiasis remain localized or get extended to include esophagitis Systemic candidiasis- C.albicans, C.glabrata & C.tropicalis three target organs ,eyes, kidneys & skin.
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VIRUSES
Herpes virus hominis (Herpes simplex)

RNA group
Paramyxo v
Rubeola(measles) Mumpsvirus

DNA group
Herpes v
Herpes simplex Varicella- zooster Cytomegalo v Epstein barr

Virus division on the basis of Picorna v viral nucleic acid, the size of virus, shape, presence of an envelope, chemical & physical nature of the virus & site of assembly
Poliovirus Coxsackievirus Aphthovirus

Pox v
(molluscum contagiosum)

Toga v
Rubellavirus

Papovirus
Papillomavirus(warts )
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Clearly, the normal bacterial flora of the oral cavity benefit from their associations with their host. Are there benefits as well to the host?

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DENTAL PLAQUE
Definition- Soft deposit that forms the biofilm adhering to the tooth surface or other hard surfaces in the oral cavity,including removable & fixed restorations.

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DENTAL PLAQUE
Classification- Supragingival / Subgingival - Adherent/Non-adherent - Cariogenic / Periodontopathic Supragingival - adherent & gram positive org.Cariogenic Subgingival less adherent & gm ve org -Periodontopathic
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DENTAL PLAQUE
Composition

Micro organisms 70-80% 1Gm 500 distinct microbial spp

Organic & inorganic components 20-30%

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FORMATION, STRUCTURE AND ORGANISATION OF DENTAL PLAQUE

Acquiring the pellicle. Transport of microorganisms. Long range physicochemical interactions. Short range interactions. Co aggregation of microorganisms. Multiplication of attached organisms.
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DENTAL PLAQUE
Pellicle formation- Distinct organic structure on surface of teeth prior to colonisation by bacetria,consists of glycoproteins derived from saliva Thickness-100nm-2hrs -500-1000nm-24hrs

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 Early plaque principally composed of cocci Salivary pellicle-Specific attachment site Increase in size by multiplication than by apposition Surface of early plaque-rod shaped bacteria-next phase filamentous bacteria

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 Streptococci and Actinomyces are early colonizers S.sanguis is the first to be discovered

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Secondary colonisersPrevotella,Capnocytop haga spp,F.nucleatum,P.gin givalis Coaggregation

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One day old plaque

Non-bacterial components
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Developed Supragingival plaque

Long Standing supra gingival plaque

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MICROBIAL COMPOSITION OF DENTAL PLAQUE IN HEALTH

Bacterium
Streptococcus Actinomyces Lactobacilli Veilonella Fusobacterium Spirochetes Gm ve Anaerobes

Fissures
++++ +++ +/++ +/-

Approximal
+++ ++++ +/+++ + +

Gingival Crevice
+++ +++ +/++ ++ + ++
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HYPOTHESIS RELATING PLAQUE TO CARIES Specific plaque hypothesis: only a very limited number of species are involved in disease. -specific pathogens. Non-specific plaque hypothesis: plaque is a microbial community. Ecological plaque hypothesis: shifts in the balance of the resident plaque microflora.
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ECOLOGIC PLAQUE HYPOTHESIS

EXCESS SUGAR

NEUTRAL pH

S. sanguis S.oralis

REMINERALISATION

STRESS

ENVIRONMENTAL CHANGE

ECOLOGICAL SHIFT

DISEASE

ACID PRODUCTION

LOW pH

MS LACTOBACILLI

DEMINERALISATION

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BACTERIA AND DENTAL CARIES

Dental caries- Slow decomposition of teeth resulting from loss of hydroxy apatite crystals
Plaque

Host

Substrate

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PATHOGENIC DETERMINANTS OF CARIOGENIC BACTERIA

Rapid transport of dietary sugars: Mutans streptococci possess more than one sugar transport system. Rapid rates of glycolysis (acidogenicity): can result in a terminal pH of below 4.5 in only a few minutes. Tolerance of, and growth at, low pH (aciduricity): the growth of many of the bacteria found on sound enamel (e.g.. Strep.

Sanguis) is inhibited at pH <5.5, whereas this is optimal for

cariogenic species.
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 Extracelluar polysaccharide synthesis (EPS): these polymers help make up the plaque matrix. Glucosyltransferases (GTF's) convert sucrose to soluble and insoluble glucans, that help consolidate bacterial attachment. Fructosyltransferases (FTF's) convert sucrose to fructans; these polymers are labile and can be used by plaque bacteria as an energy source.
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Intracellular polysaccharide synthesis (IPS): can be used during starvation conditions and catabolised to acid when dietary sugars are not available. mutans streptococci but not lactobacilli produce EPS.

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PATHOGENIC PROPERTIES OF CARIOGENIC BACTERIA

Rapidly transport fermentable sugars when in competition with other bacteria Conversion of such sugars to acid. The production of extra-cellular and intra- cellular polysaccharides. The ability to maintain sugar metabolism under extreme environmental conditions.
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103

Prior to 1890 it was believed that mineral acids that formed from food residues were responsible for tooth decay and there was no connection with bacteria.

1890 - W.D. Miller formed the chemo parasitic theory of caries .Microorganisms attach to the tooth and produce organic acids which dissolve the enamel.

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A diet that could produce caries in normal animals could not cause caries in germfree animals. caries was proven to be caused by bacteria.

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Evidence of Bacterial Role:

Germ free animals do not develop caries. Antibiotics fed to animals are effective in reducing the caries rate. Totally unerupted and unexposed teeth do not develop caries. Oral Bacteria can demineralize enamel and dentin and produce caries like lesions. Microorganisms have been histologically demonstrated invading carious enamel and dentin.

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1924 - Clarke isolated a streptococcus from approx. 72% of carious lesions. These formed rods in old cultures and therefore he called it S. mutans.

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 1933 - Tomato juice agar was used to culture oral lactobacilli. Higher numbers were found in mouths with caries Similar results with other media selective for the lactobacilli gave rise to the belief that lactobacilli were the cause of tooth decay.
108

Gnotobiotic when inoculated with enterococci, could get caries.

Therefore, not just lactobacilli but other organisms were also implicated!
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 1967 & 1968 - Carlsson & Edwardson worked on the taxonomy of the various cariogenic streptococci

They named the cariogenic streptococci "Streptococcus mutans

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The characteristics which distinguished the S. mutans from all other streptococci were their ability to: -ferment mannitol and sorbitol -produce insoluble glucan from sucrose

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Great Lake Study

In the 1970s, DRG, in Great Lakes, Illinois Naval Training Center studied S. mutans-free and caries-free individuals. -60% of them became infected -Disproved genetic basis for caries resistance - Therefore caries was shown to be a communicable disease for humans
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WOMB TO TOMB STUDY

Carlsson & collegues-Two phased study

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WINDOW OF INFECTIVITY OF S. mutans

-Carfield(1993). -6-12yrs
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 Caries can be divided into Enamel, Dentinal & Root surface (cemental) caries Enamel caries further divided into Smooth Surface Pit & Fissure
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 Smooth Surface most consistent are gram positive facultative cocci, S.mutans & S.salivarius. S.mutans primary etiologic agent Role of S.salivarius in caries production is not well known
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Normal Buccal-Lingual smooth surface caries - S.

mutans group
Normal Interproximal Smooth Surface caries - S.

mutans and lactobacilli

Rampant Smooth Surface caries- S. mutans group

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 Pit & Fissure caries- Wide

variety is found because of the varied environment present S.mutans & Lactobacilli are suspected etiologic agents. Anaerobic rods, Bifidobacterium, Eubacterium, & Propionbacterium identified. Actinomyces,& Bacillus sp.- invasive front of the deep dentinal lesions
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Root surface lesion High no. of Actinomyces sp. including A .viscosus, A.naeslundii & A. odontolyticus.

119

Rampant Caries- Any tooth surface. characterized by a rapid dissolution of the tooth It is not due to different organisms

The rapid progression is due to the frequency of ingestion of sucrose or other fermentable sugar!
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Endodontic infections
Vital dental pulp-Sterile Endodontics disease of pulp & periapical tissues W.D.Miller- bacterial involvement Object of many errors Evidence experiments on germ free rats.
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Bacterial Ecology Of The Infected Root Canal

Microbial composition determined by a) the root by which bacteria gain access to the root canal & b) the no. & quality of ecological factors

Bacteria are in the dynamic state influenced by the 1)interactions bet. bacteria & 2) interaction with the host

Root canal is much less complex than the Subgingival flora

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 Experiments Initially facultative species of Streptococcus, Enterococcus ,Lactobacillus, Corynebacterium & Coliform rods, anaerobic organisms, Peptostreptococcus, Propionibacterium, Eubacterium,Prevotella & Fusobacterium. Proportion of anaerobic bacteria increased with time outnumbering the facultative species. Greater anaerobes in the apical region than main canal.

123

 Apical region Bacteria are fastidious have strict nutritional req. Serum nutrients-Apical region Proteolytic bacteria Prevotella, Porphyromonas, Peptstrepto, Fusobacterium main group Staphylococci,Strept. ,Lacto, & Neisseria opt main canal.

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Main canal- metabolic activity is very low.

Accidental entry of instruments- periapex blood entry can serve as nutrient for bacteria this flares up the host response and Acute apical periodontitis results.
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Periapical abscess- 75% anaerobes.

3 to 4 bacterial isolates-Porphyromonas, & Prevotella are the common isolates. Peptostrepto.-Second most frequent. Viridans strept.-Third most common. Veilonella, Eubacterium, Actinomyces, Lactobacillus & Fusobacterium are also isolated.
126

Periodontal infections
Periodontal disease- No. of distinct clinical entities
affecting Periodontium Chronic bacterial infections Nature of disease-bacterial host interactions Host response-protective, destructive, or both

127

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 Identification of bacterial etiology1. Large no bacteria ass. with disease 2. Elimination reverses the disease 3. Elevated host responses assoc. With disease 4. Pathogenicity similar to periodontal disease occurs with implantation into germ free animals 5. Bacteria possess potentially pathogenic mediators

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Gingival health - Harbours microflora in health which is simple & sparse ,flora similar to early stages of plaque formation Gram positive- majority- constitute two thirdsFilamentous forms are also seen Disease- are a result of one or two mechanisms direct effect include invasion, exotoxins, cell constituents & enzymes, indirect response are immunologic & host responses
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Localized juvenile periodontitis

Clinical-12-20 yrs No/little gingival inflammation Marked, localized alv. bone loss perm 1st molar & incisors Microbiology-cultural studiesgram negative rods, Actinobabacillus, Actinomycetemcomitans.
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 Generalized juvenile periodontitis

Clinically-similar to LJP Microbial findings- B.gingivalis most common 1320%,Eikenella.corrodens,B.intermedius, Capnocytophaga, Neisseria. A. actinomycetemcomitans in low no.

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Pre-pubertal periodontitis
Clinically- A rare form of priodontal disease occurs during or immediately after the eruption of the primary teeth. Females. Generalized & localized forms Microbiolgic findings- Fusobacterium, Wolinella, Bacteroides & Capnocytophaga commonly found
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ANUG (Acute Necrotizing Ulcerative Gingivitis)

Clinically-Marginal gingiva Young people under stress & immunocompromised pts. Necrotic lesions of one or more interdental papilla & progress to maximal extent associ.with pain, bleeding, fetor exore & fever Cancrum oris (NOMA) Considerably beyond gingiva & can give rise to life threatening infections.
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ANUG Microbiology - Harbour high no. of Spirochetes & P.intermedia Early microscopic examination identified high levels of Fusobacteria (gram negative rods)

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Four Histopathological zones1)Bacterial zone containing variety of bacteria similar to sub gingival flora 2)Neutrophil zone, rich in leukocytes 3)Necrotic zone-Spirochetes & rod shaped bacteria 4)Zone of spirochetal infiltration-inter.& large spirochetes b/w normal collagen
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REFERENCES -Oral Microbiology and ImmunologyNewman & Neisengard -Cariology-Newbrun -Contemporary oral microbiology & immunology-Jogren Slots -J.Nihon Univ.Sch.Dent.Vol.36,No.1,133,1994 -Incipient & hidden caries oct 2005 49:4 -Text book of microbiology-Ananth

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