Oral microbiology- Dr Harshavardhan Patwal
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Transcript of Oral microbiology- Dr Harshavardhan Patwal
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Dr Harshavardhan G Patwal
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INTRODUCTION
HISTORY
NORMAL MICROFLORA OF HUMAN ORAL CAVITY
MICROFLORA OF PERIODONTAL DISEASE
VIRULENCE FACTORS
BACTERIA IN PATHOGENESIS OF PERIODONTAL
DISEASE
MICROBIAL DIAGNOSIS
CONCLUSION
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INTRODUCTION
• FROM AMOEBA TO Aa, Pg
• BACTERIAL PLAQUE- CAUSATIVE OF
PERIODONTAL DIS
• LARGE COMPLEX MIXTURE OF BACTERIAL
SPECIES
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HISTORY• 1683 - ANTON VON LEEWENHOEK
• 1890 - WD MILLER
• 1965 - LOE, THEILADE AND JENSEN
• 1880 TO 1890 – AMOEBAE, SPIROCHETES,
MILLER’S BACTERIA, STREPTOCOCCI
• 1890 – 1930 SPECIFICITY THEORY
• 1930 – 1970 NONSPECIFICTY THEORY
• 1970 ONWARD RETURN OF SPECIFICITY THEORY
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NORMAL FLORA• 300 – 400 SPECIES FOUND IN HUMAN
SUBGINGIVAL PLAQUE
• INDIGENOUS FLORA
• SUPPLEMENTAL FLORA
• TRANSIENT FLORA
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FACTORS INFLUENCING NORMAL FLORA• THE ENVIRONMENT THE INDIVIDUAL IS RAISED
• THE DIET
• ORAL HYGIENE HABITS
• INFECTION POTENTIAL FROM FAMILLY MEMBERS
• INFECTION POTENTIAL FROM PETS
• TOOTH ERUPTION
• PRESENCE OF PERIODONTAL POCKET
• HORMONAL EFFECTS
• PERIODIC EXPOSURE TO VARIOUS TREATMENT
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ACQUISITION OF BACTERIA
• STARTS AT BIRTH
• DERIVED FROM FROM THE IMMEDIATE
ENVIRONMENT
• MODULATED BY HOST FACTORS
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SUBGINGIVAL FLORA ASSOCIATED WITH HEALTH
• STREPTOCOCCUS MITIS
• S SANGUIS
• S EPIDERMIDIS
• ROTHIA DENTOCARIOSA
• ACTINOMYCES VISCOSUS
• ACTINOMYCES NAESLUNDII
• SMALL SPIROCHETES
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RELATIVE PROPORTION OF MICROORGANISMS
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MICROBIAL FLORA OF PERIODONTAL DISEASE
CONCEPTS OR HYPOTHESISES – WALTER LOESCHE 1976
NONSPECIFIC – THE ENTIRE PLAQUE FLORA IS
RESPONSIBLE
SPECIFIC – ONLY CERTAIN PLAQUE IS PATHOGENIC
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RANKING OF SUSPECTED BACTERIAL PATHOGENS
MODERATE• S intermedius• P micros• F nucleatum• E corrodens• Eubacterium
STRONG• P intermedia• C rectus• E nodatum• Treponema sp
VERY STRONG• A actinomycetemcomitans• P gingivalis• B forsythus
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VIRULENCE FACTORS
VIRULENCE - THE QUALITY OF BEING DAMAGING;
THE DISEASE PRODUCING POWER OF A
MICROORGANISM
GLOSSARY OF PERIODONTAL TERMS
1984
OTHER FACTORS :
CONCENTRATION OF MICROORGANISMS
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VIRULENCE FACTORS
1. THE CAPACITY TO COLONIZE
2. THE ABILITY TO EVADE ANTIBACTERIAL HOST DEFENSE
3. THE ABILITY TO PRODUCE SUBSTANCES THAT CAN
DIRECTLY OR INDIRECTLY INITIATE TISSUE
DESTRUCTION
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COLONIZATION
FACTORS INFLUENCING COLONIZATION
1. AVAILABILTY OF SURFACES TO ATTACH
2. ABILITY TO MULTIPLY
3. ABILITY TO COMPETE SUCCESSFULLY AGAINST
OTHER MICROORGANISMS
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ATTACHMENT
SITES FOR ATTACHMENT1. THE TOOTH2. THE SULCULAR / POCKET EPTHELIUM3. OTHER BACTERIAL SPECIES
BACTERIAL FACTORS FACILITATING ATTACHMENT4. ADHESINS5. COAGGREGATION
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ADHESINS1. FIMBRIAE
a. TYPE --1b. TYPE –2c. TYPE –3d. TYPE – 4
2. CELL ASSOCIATED PROTEINSa. 300-330 kDa PROTEINb. 70, 90 kDa PROTEINc. 40 kDa PROTEIN
RECEPTORS FOR ADHESINS ON TISSUE SURFACES• GALACTOSYL VESICLES• SIALIC ACID• PROLIN-RICH PROTEINS OR STATHERIN• TYPE-I AND IV COLLAGEN
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FIMBRIAE
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ADHESINS ORGANISM
FIMBRIAE E corrodensA viscosus
PROTEINS
F nucleatum (300-330 kDa protein)T denticolaP gingivalisStreptococcus mitis (70, 90 kDa protein)B loeschei ( fimbriae associated 75 & 45
kDa polypeptide)
EXAMPLES OF ADHESINS
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RECEPTOR ADHESINS SITE ORGANISMS
GALACTOSYL RESIDUE
PROTEINFIMBRIAE
EPITHELIAL CELLSFIBROBLASTSPMLSALIVA COATED SURFACES
E corrodens (PROTEIN)B gingivalis (FIMBRIAE)A viscosus (TYPE 2 FIMBRIAE)A naeslundii (FIMBRIAE)T denticola (PROTEIN)F nucleatum (300-330 kDa Protein)B loeschi (FIMBRIAE)
SIALIC ACID PROTEIN SALIVA COATED SURFACES
S mitis (70, 90 kDa PROTEIN)
PROLIN-RICH PROTEINS OR STATHERIN
FIMBRIAE MINERALIZED SURFACES A viscosus (TYPE 1 FIMBRIAE)
TYPE-I AND IV COLLAGEN
BASEMENT MEMBRANE B gingivalis ( NOT KNOWN)
MANNOSE RESIDUE PROTEIN FIBROBLASTS T denticola (PROTEIN)
SALIVA TREATED HYDROXYAPPATITE
FIMBRIAE MINERALIZED SURFACES A viscosus (TYPE 1 FIMBRIAE)
KNOWN RECEPTORS FOR BACTERIAL ADHESINS
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COAGGREGATION
Sl. No. TARGET SPECIES RECEPTOR ON TARGET SPECIES
SPECIES ADHESINS
1 B gingivalis GALACTOSYL RESIDUE
F nucleatum PROTEIN
2 S sanguis GALACTOSYL RESIDUE
B loeschi FIMBRIAE ASSOCIATED 75 AND 45 kDa POLYPEPTIDE
3 S sanguis GALACTOSYL RESIDUE
A viscosus TYPE 2 FIMBRIAE
4 S sanguis POLYSACCHARIDE WITH REPEATING HEPTASUCCINATE
A viscosus FIMBRIAE
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MULTIPLICATION OF BACTERIA
FACTORS INFLUENCING MULTIPLICATION
1. CREVICE TEMP
2. GCF pH
3. OXIDATION REDUCTION POTENTIAL
4. AVAILABLE NUTRIENTS
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INTERBACTERIAL RELATIONSHIP• FAVORABLE RELATIONSHIP
• FACILATITING ATTACHMENT
• PRODUCING GROWTH FACTORS
– BRANCED CHAIN FATTY ACIDS
– POLYAMINES
– VITAMIN K ANALOGUES
– LACTATE / FORMATE /HYDROGEN
• PRODUCTION OF EXTRACELLULAR ENZYMES THAT
UNCOVER BINDING SITES FOR ONOTHER
ORGANISM
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ANTAGONISTIC RELATIONSHIP
• COMPETITION FOR NUTRIENTS
• COMPETITION FOR SITE
• PRODUCTION OF SUBSTANCES LIMITING /
PREVENTING GROWTH
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INTERRELATIONSHIP FOR NUTRITIONO2
SUGARCO2
SALIVA DIET
GCFSALTS
NH4
AMINO ACIDS
STRPTOCOCCUSACTINOMYCES
PABAS MUTANS
CAPNOCYTOPHAGAE CORRODENSAa
FUSOBACTERIUMEUBACTERIUMBACTEROIDSPEPTOCOCCUSPEPTOSTREPTOCOCCUSA
Thiamin pyroPPutrescineisobutyrate
CO2NH4
T macrodentium
lactate
Veillonella acetate
Eubacterium
H2
CampylobacterWolinella
formate
P melaninogenicusP gingivalis
Hemin
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OVERCOMING HOST DEFENSESl. No DEFENSE
MECHANISMVIRULENCE FACTOR SPECIES
1 SPECIFIC AbIgA, IgG PROTEASE B gingivalis
B intermediaB melaninogenicusCapnocytophaga
2 SALIVARY MACROMOLECULES
AFFINITY FOR HIDDEN RECEPTORS (CRYTITOPES)
A viscosus
3 PNL
LEUKOTOXIN Aa
CAPSULE B gingivalis
INHIBITION OF DEGRANULATION T denticola
INHIBITION OF SUPEROXIDE PRODUCTION Treponema
4LYMPHOCYTE
LEUKOTOXIN – LETHAL Aa
LEUKOTOXIN – NONLETHAL SUPPRESION Aa
SUPPRESSED IMMUNE RESPONSE B intermedia
INHIBIT RESPONSE TO MITOGENS AND ANTIGENS
B intermediaB endodontalisT denticolaAa
5 ANTIBIOTIC THERAPY
CONJUGAL TRANSFER OF RESISTANCE PLASMID
T denticolaB intermedius
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FACTORS CAUSING TISSUE DAMAGESl. No. ENZYMES SPECIES
1 COLLAGENASE P gingivalisAa
2 PROTEASE P gingivalisP intermedia
3 KERATINASE P gingivalisT denticola
4 ARYL SULFATASE W recta
5 NEURAMINADASE P gingivalisB forsythusP melaninogenicus
6 FIBRONECTIN DEGRADING ENZYMES
P gingivalisP intermedia
7 PHOSPHLIPASE P intermediaP melaninogenicus
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BACTERIAL PREPARATION INHIBITING MAMMALIAN CELL GROWTH
Sl. No. BACTERIAL PREPARATIONS ORGANISM AFECTED CELLS
1 CULTURE SUPERNATANTS P gingivalis FIBROBLASTS
2 LPS Aa FIBROBLASTS
3 CAPSULE Aa FIBROBLASTS
4 SONIC EXTRACTS P gingivalisT denticola
FIBROBLASTS
5 SONIC EXTRACTS AaP gingivalis
EPITHELIAL CELLSENDOTHELIAL CELLS
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Environmental and acquired risk factor
Host im
muno-inflam
matory
response
Ct &
bone metabolism
Clinical signs of disease
initiation & progression
Genetic risk factor
Cytokines
MMPs
Ab
PMN s
Ag
LPS
Othervirulence factors
Microbial challenge
MECHANISM OF PERIODONTAL DISEASE PRODUCTION
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BACTERIA ASOCIATED WITH SPECIFIC PERIODONTAL DISEASE
DISEASE ORGANISM
ADULT PERIODONTITIS P gingivlis, P intermedia, B forsythus, C rectus, E corrodens, F nucleatum,
REFRACTORY PERIODONTITIS B forsythus, P gingivlis, C rectus, P intermedia
LJP Aa, Capnocytophaga
PRAGNENCY GINGIVITIS P intermedia
ANUG P intermedia, Intermediate-sized spirochetes
PERIODONTITIS IN JUVENILE DIABETICS
Capnocytophaga, Aa
PERIODONTAL ABSCESSES P gingivalis, Fusobacterium, Capnocytophaga, Vibrio sp
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MICROBIAL DIAGNOSIS OF PD
• MICROSCOPIC ASSAYS
• CULTURE ASSAYS
• IMMUNOLOGIC ASSAYS– ELISA
– IMMUNOFLUORESCENCE
• DNA PROBE ASSAY
• ENZYME-BASED ASSAYS
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AVAILABLE DIAGNOSTIC AIDS• CULTURE
Aa P gingivalis
P intermedia Capnocytophaga
Wolinella Sp Eikenlla Sp
Peptostrptococcus micros
• DNA PROBE
Aa B forsythus
P gingivalis C rectus
• BANA
T denticola B forsythus
P gingivalis
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CONCLUSION• COMPLEXITY IS THE PRINCIPAL PROBLEM
• REMARKABLE RESEARCH EFFORTS HAVE
RESULTED IN UNDERSTANDING THE MICROBIAL
ETIOLOGY
• MORE WORK REQUIRED TO DEVELOP
DIAGNOSTIC TESTS
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