microbiology of dental caries introduction infection: disease: asymptomatic carriage: ...
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BACETRAIL PATHOGENESIS
What is virulence?
The ability of a bacterium to cause infection.
Virulence factors: Two types: Those that promote bacterial colonization and invasion
of the host tissue Those that cause damage of the host tissue.
Research in the past four decades have accumulated information which led to identification of possible
pathogens of human dental caries.
Q. How a cause and effect relationship is established between bacterium and the disease?
A. Koch’s Postulate (1800s).
KOCH’S POSTULATES The bacterium should be found in people with the
disease The bacterium should be isolated from the lesions
of infected person Pure culture, inoculated into a susceptible
individuals or animals should produce the disease Same bacterium should be re-isolated from
intentionally infected animals or humans.
LIMITATIONS OF KOCH’S POSTULATES
Virulence is within the bacterium and is independent of the host
Isolation and growth of bacterium is necessary: Yet, some pathogens not yet cultured
Nos. 2 & 4: assume that all members of the same species are virulent
No. 3: Ethics with human subjects, Yet some pathogens from humans can not cause the same effect in animals.
WHAT IS THE ALTERNATIVE?
MOLECULAR POSTULTES1. Gene should be found in the bacterial strain.
2. Disturbing the virulent gene should reduce its virulence.
3. Bacterial virulent gene should be expressed in the animal or human at sometime during the infectious process
4. Abs to gene product should be protective or should elicit protective immunity (cell-mediated).
Q. Why Did it Take Long Time for Caries Microbiology?
Complex ecology of the oral cavity.300 – 400 species are indigenous oral flora.History:
Miller (1880): Little knowledge about which bacteria. Clarke (1924): First who associate bacteria with dental caries
o First to isolate MS from human dental carieso First to produce caries in extracted teeth.
Orland (1955): Used animals to induce dental caries using MS.
DENTAL CARIES MICROBIOLOGY RESEARCH
1960s : germ-free animals 1960s and 70s: importance of glucan (glucanase):
Clinical trials using glucan hydrolyase rinsesGlucan is plaque enhancerProblem with isolation: Number, media ….etc.Specific plaque theory
MS identified as an associated bacteria with caries.
MUTANS STRPETOCOCCI (MS) TYPES: (Coykendall, 1989)
S. anginosus : important in purulent infections S. bovis : found in patients with colon cancer S. mitis : similar to sanguis but doesn’t ferment any sugar S. mutans : seven species S. salivarius : in saliva, rare in infections S. sanguis : causes endocarditis S. vestbularis : new species from oral cavity.
STREPTOCOCCUS MUTANS
Species Serotype Arg Raf Mel H2O2 Aero Baci Source
S mutans c, e, f - + + - + - Human
S rattus b + + + - + - Rats
S cricetus a - + + - - + Rats
S sobrinus d, g - - - + + - Human
S ferus c - - - - - + Rats
S macacae c - + - - - + Monkey
S downei h - - - - - + Monkey
WHY S. mutans SUCCEED?
Three factors: Ability to adhere to other bacteria and tooth surface Ability to rapidly metabolize nutrients (CHO) Ability to tolerate acidic environment.
ADHERENCE OF S. mutans Saliva:
Lysozyme IgA: (IgA protease), (IgA deficiency)
Bacterial proteins: Ag I/II family: Adhere to saliva proteins Adhesin Fimbrial adhesion: Adhere to saliva pellicle glucan binding (GBP)
CHO METABOLISM BY S.mutans
CHO must be transported across the membrane (Sugars must be phosphorylated):
Multiple Sugar Metabolism (MSM) System:
Transport via the Phosphoenolpyruvate (PEP):
Sugar Phosphotransferase System (PTS):
CHO Metabolism
PEP + CHO PTS Pyruvate +P-CHO
S.mutans enolase: Fluoride inhibits it. S.mutans store polysaccharides .. Why?
S. mutans ACID TOLERANCE
Through cell membrane, extrusion of protons: Membrane ATPase hydrolyze ATP molecules Hydrolysis of one ATP, results in extrusion of three
protons This results in elevation of cytoplasmic pH. When pH decreases, ATPase activity increases 4-folds.
COLONIZATION OF S. mutans
Based on ability of S. mutans to synthesize insoluble glucan.
S. mutans have 3 genes: gtfB encodes GTF-I enzyme: insoluble glucan gtfC encodes GTF-SI enzyme: insoluble glucan gtfD encodes GTF-S enzyme: soluble glucan
RESEARCH USING GTFs
Purified S.mutans GTFs were used for caries immunization in rodents.
(Smith et al., 1979).
Implantation of S. mutans defective in IS glucan synthesis into rats resulted in reduced smooth surface caries induction.
(Munro et al., 1991).
Strain Gtase Adherence%
MT8148 I,SI/S 72.8 2.6
B29 /SI/S 16.3 1.0
B29 I/SI/S 46.9 5.9
B58 I/ /S 9.6 1.0
B58 I/SI/S 69.9 1.8
B32 / /S 1.4 0.4
(Fujiwara et al., 1996)
ACQUISITION OF S. mutans Sterile mouth at birth
S. sanguis and S. mutans colonize teeth
Number of bacteria increases in the presence of: Sucrose Caries Teeth
ACQUISITION OF S. mutans
Birth 5 YearFirst Tooth 19 336.8 +/- 1.4 mo.
26
MS
N=38
Caufield et al., J Dent Res. 72:37-45, 1993.
ACQUISITION OF S. mutans
Important facts: Difficult to change S. mutans strain(s) High number of S.mutans strains and isolates. One (or more) strain (isolates) is/are present in the
mouth.
TRANSMISSION OF CARIOGENIC FLORA
Mothers to children: DNA technique Method of transmission
Spouses: Different bacteria studied
Replacement Therapy
IF WE UNDERSTAND THE DENTAL CARIES
MICROBIOLOGY WELL..
WE WILL TREAT PATIENTS
DIFFERENTLY !!!!
HOW?????????????????????????????