Oral microbiology- Dr Harshavardhan Patwal
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This document discusses the microflora associated with periodontal disease. It begins with an introduction to bacteria's role in periodontal diseases and provides a history of research in this area from the 17th century to present. It then describes the normal oral microflora and microflora associated with periodontal disease. Key pathogens implicated in periodontal disease include Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. The document also discusses virulence factors of periodontal bacteria and their role in colonization, evading host defenses, and tissue destruction. Microbial diagnosis of periodontal disease is also covered.
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Oral microbiology- Dr Harshavardhan Patwal
- 1. Dr Harshavardhan G Patwal
- 2. INTRODUCTION HISTORY NORMAL MICROFLORA OF HUMAN ORAL CAVITY MICROFLORA OF PERIODONTAL DISEASE VIRULENCE FACTORS BACTERIA IN PATHOGENESIS OF PERIODONTAL DISEASE MICROBIAL DIAGNOSIS CONCLUSION
- 3. INTRODUCTION • FROM AMOEBA TO Aa, Pg • BACTERIAL PLAQUE- CAUSATIVE OF PERIODONTAL DIS • LARGE COMPLEX MIXTURE OF BACTERIAL SPECIES
- 4. 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
- 5. NORMAL FLORA • 300 – 400 SPECIES FOUND IN HUMAN SUBGINGIVAL PLAQUE • INDIGENOUS FLORA • SUPPLEMENTAL FLORA • TRANSIENT FLORA
- 6. 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
- 7. ACQUISITION OF BACTERIA • STARTS AT BIRTH • DERIVED FROM FROM THE IMMEDIATE ENVIRONMENT • MODULATED BY HOST FACTORS
- 8. SUBGINGIVAL FLORA ASSOCIATED WITH HEALTH • STREPTOCOCCUS MITIS • S SANGUIS • S EPIDERMIDIS • ROTHIA DENTOCARIOSA • ACTINOMYCES VISCOSUS • ACTINOMYCES NAESLUNDII • SMALL SPIROCHETES
- 10. MICROBIAL FLORA OF PERIODONTAL DISEASE CONCEPTS OR HYPOTHESISES – WALTER LOESCHE 1976 NONSPECIFIC – THE ENTIRE PLAQUE FLORA IS RESPONSIBLE SPECIFIC – ONLY CERTAIN PLAQUE IS PATHOGENIC
- 11. 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
- 12. 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
- 13. 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
- 14. COLONIZATION FACTORS INFLUENCING COLONIZATION 1. AVAILABILTY OF SURFACES TO ATTACH 2. ABILITY TO MULTIPLY 3. ABILITY TO COMPETE SUCCESSFULLY AGAINST OTHER MICROORGANISMS
- 15. ATTACHMENT SITES FOR ATTACHMENT 1. THE TOOTH 2. THE SULCULAR / POCKET EPTHELIUM 3. OTHER BACTERIAL SPECIES BACTERIAL FACTORS FACILITATING ATTACHMENT 1. ADHESINS 2. COAGGREGATION
- 16. ADHESINS 1. FIMBRIAE a. TYPE --1 b. TYPE –2 c. TYPE –3 d. TYPE – 4 2. CELL ASSOCIATED PROTEINS a. 300-330 kDa PROTEIN b. 70, 90 kDa PROTEIN c. 40 kDa PROTEIN RECEPTORS FOR ADHESINS ON TISSUE SURFACES • GALACTOSYL VESICLES • SIALIC ACID • PROLIN-RICH PROTEINS OR STATHERIN • TYPE-I AND IV COLLAGEN
- 17. FIMBRIAE
- 18. ADHESINS ORGANISM FIMBRIAE E corrodens A viscosus PROTEINS F nucleatum (300-330 kDa protein) T denticola P gingivalis Streptococcus mitis (70, 90 kDa protein) B loeschei ( fimbriae associated 75 & 45 kDa polypeptide) EXAMPLES OF ADHESINS
- 19. RECEPTOR ADHESINS SITE ORGANISMS GALACTOSYL RESIDUE PROTEIN FIMBRIAE EPITHELIAL CELLS FIBROBLASTS PML SALIVA 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
- 20. 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
- 21. MULTIPLICATION OF BACTERIA FACTORS INFLUENCING MULTIPLICATION 1. CREVICE TEMP 2. GCF pH 3. OXIDATION REDUCTION POTENTIAL 4. AVAILABLE NUTRIENTS
- 22. 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
- 23. ANTAGONISTIC RELATIONSHIP • COMPETITION FOR NUTRIENTS • COMPETITION FOR SITE • PRODUCTION OF SUBSTANCES LIMITING / PREVENTING GROWTH
- 24. INTERRELATIONSHIP FOR NUTRITION O2 SUGAR CO2 SALIVA DIET GCF SALTS NH4 AMINO ACIDS STRPTOCOCCUS ACTINOMYCES PABA S MUTANS CAPNOCYTOPHAGA E CORRODENS Aa FUSOBACTERIUM EUBACTERIUM BACTEROIDS PEPTOCOCCUS PEPTOSTREPTOCOCCUSA Thiamin pyroP Putrescine isobutyrate CO2 NH4 T macrodentium lactate Veillonella acetate Eubacterium H2 Campylobacter Wolinella formate P melaninogenicus P gingivalis Hemin
- 25. OVERCOMING HOST DEFENSE Sl. No DEFENSE MECHANISM VIRULENCE FACTOR SPECIES 1 SPECIFIC Ab IgA, IgG PROTEASE B gingivalis B intermedia B melaninogenicus Capnocytophaga 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 4 LYMPHOCYTE LEUKOTOXIN – LETHAL Aa LEUKOTOXIN – NONLETHAL SUPPRESION Aa SUPPRESSED IMMUNE RESPONSE B intermedia INHIBIT RESPONSE TO MITOGENS AND ANTIGENS B intermedia B endodontalis T denticola Aa 5 ANTIBIOTIC THERAPY CONJUGAL TRANSFER OF RESISTANCE PLASMID T denticola B intermedius
- 26. FACTORS CAUSING TISSUE DAMAGE Sl. No. ENZYMES SPECIES 1 COLLAGENASE P gingivalis Aa 2 PROTEASE P gingivalis P intermedia 3 KERATINASE P gingivalis T denticola 4 ARYL SULFATASE W recta 5 NEURAMINADASE P gingivalis B forsythus P melaninogenicus 6 FIBRONECTIN DEGRADING ENZYMES P gingivalis P intermedia 7 PHOSPHLIPASE P intermedia P melaninogenicus
- 27. 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 gingivalis T denticola FIBROBLASTS 5 SONIC EXTRACTS Aa P gingivalis EPITHELIAL CELLS ENDOTHELIAL CELLS
- 28. Environmental and acquired risk factor Hostimmuno-inflammatory response Ct&bonemetabolism Clinicalsignsofdisease initiation&progression Genetic risk factor Cytokines MMPs Ab PMN s Ag LPS Other virulence factors Microbial challenge MECHANISM OF PERIODONTAL DISEASE PRODUCTION
- 31. 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
- 32. MICROBIAL DIAGNOSIS OF PD • MICROSCOPIC ASSAYS • CULTURE ASSAYS • IMMUNOLOGIC ASSAYS – ELISA – IMMUNOFLUORESCENCE • DNA PROBE ASSAY • ENZYME-BASED ASSAYS
- 33. 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
- 34. CONCLUSION • COMPLEXITY IS THE PRINCIPAL PROBLEM • REMARKABLE RESEARCH EFFORTS HAVE RESULTED IN UNDERSTANDING THE MICROBIAL ETIOLOGY • MORE WORK REQUIRED TO DEVELOP DIAGNOSTIC TESTS