By Kate Egge
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• Background information about bad breath and
Periodontitis
• Discuss 3 bacteria that cause oral malodor
• Methods for detecting bacteria and?or Bad breath:
PCR, organoleptic, VSC concentrations, and BANA tests
• Research: that indicates bacteria cause bad breath
• Research: that indicates that bacteria may cause periodontitis
• Conclusions
• References
• Questions
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• Americans spent 625 million dollars on breath fresheners in
2000.
• 85-95% of cases originate in the mouth
• Microorganisms on the posterior dorsal side of tongue is the main source of bad breath
• Area is poorly cleansed by saliva and contains invaginations where bacteria can hide and putrefy postnasal drip
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• Some bad breath is associated with periodontal disease and destruction of gums
• Periodontitis is a common, progressive disease that affects the supporting structures of the teeth, causing loss of attachment to the bone and often resulting in tooth loss
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• Bad breath primarily result of microbial metabolism.
• Organisms feed off of glycoproteins and release offensive chemical odors
• Oral gram-negative organisms such as spirochetes and some bacteroides have the capacity to produce oral malodor through putrification of sulfur containing protein substrates such as cysteine and methionine
• Chemistry
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• Key anaerobic organisms
,
and
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• Gram negative, anaerobic spirochete
• A lot of size variation among the spirochetes
• Form a major phylogenetic lineage of Bacteria
• Protoplasmic cylinder
• Flagella emerge from each pole and are located in the periplasm of the cell (endoflagella)
• Endoflagella and protoplasmic cylinder are surrounded by a flexible membrane called the outer sheath
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Porphyromonas gingivalis
Bacteroides forsythus
• Member of genus Bacteroides
• Gram negative obligate anaerobe
• Ferments sugars to primarily acetate and succinate
• Bacteroides species dominant bacteria in large intestine, found 10^10 – 10^11 cells per gram of human feces
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Porphyromonas gingivalis
• Several clinical studies show that
is a factor in periodontitis
• Study Journal of Clinical Microbiology compared the prevalence of
a group with periodontitis to a different group that was periodontally healthy in
• Results:
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Porphyromonas gingivalis
•
was detected in only 25% of the healthy subjects but was detected in
79% of the periodontitis group
• The odds ratio for being infected with
was 11.2 times greater in the periodontits group than in the healthy group.
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Porphyromonas gingivalis
PCR
• PCR assays were used for detection of
• Target sequence was the ribosomal DNA spacer region between 16S and 23S ribosomal genes
• DNA fragments were separated by agarose gel electrophoresis
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Porphyromonas gingivalis
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• Organoleptic measurements:
• Scale 0=no odor up to 5=foul odor
• Tongue malodor was assessed by scraping the posterior tongue dorsum with gauze and immediately smelling it’s odor.
• Malodor in dental sites was assessed by pushing waxed dental floss between teeth and immediately smelling and recording the odor based on the organoleptic scale
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• High VSC concentrations indicate oral malodor
• Volatile Sulfur Compounds (VSC) are measured with a portable sulfide monitor
• VSC monitor contains a pump which sucks air from a plastic straw placed inside the subjects mouth
• The mouth air is passed through an electrolytic sensor which measures the concentration of volatile sulfurs
• VSC levels are strongly associated with organoleptic scores
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• BANA test
Test kit for bacterial trypsin like activity based on the hydrolysis of benzoyl-DL-arginine-naphthylamide (BANA) used to asses the presence of one or more of
T. denticola , P. gingivalis , B. forsythus
• These bacteria are capable of hydrolyzing the synthetic trypsin substrate BANA- an activity not identified in other oral microbial species
• Conditions favoring a Positive BANA test on the tongue dorsum and tooth sites were associated with oral malodor production
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• Species identified by BANA tests have been tentatively implicated as putative periodontal pathogens
• However, some studies have found that species
Treponema denticola , Porphyromonas gingivalis, and Bacteroides forsythus identified by BANA tests do not cause periodontitis.
• When peridontitis was present 87.5% of tooth sites and 92.5% of tongue sites were BANA (+)
• In healthy individuals 74.4% of tooth sites and
93% of tongue sites were BANA (+)
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• Oral malodor is not directly associated with presence of periodontitis
• Substantial oral malodor may be present in individuals without periodontitis
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• Gram negative anaerobic oral bacteria ( Treponema denticola ,
Porphyromonas gingivalis, and Bacteroides forsythus ) cause bad breath
• Key anaerobic organisms Treponema denticola , Porphyromonas gingivalis, and Bacteroides forsythus degrade glycoproteins proteins and release foul smelling chemicals
• Strong evidence indicates that these bacteria cause bad breath
• Treponema denticola: Anaerobic spirochete, Protoplasmic cylinder and endoflagella
• Porphyromonas gingivalis, and Bacteroides forsythus are both m embers of the genus Bacteroides
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• Bacteria on the posterior dorsal side of tongue is the main source of bad breath
• One study using PCR found that P.gingivalis was 11x more likely to be found in individuals that had periodontitis- P. gingivalis may be a periodontal pathogen
• However, research using BANA tests found that the presence of T. denticola , P. gingivalis, and B. forsythus was NOT associated with periodontitis
• Oral malodor is not directly associated with the presence of periodontitis
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• Rosenberg, Mel. Clinical Assessment of Bad Breath: Current Concepts.
Journal of the American Dental Association 1996; 127: 475-82
• Griffen A, Becker M, Lyons S. Prevalence of Porphyromonas gingivalis and
Periodontal Health Status. Journal of Clinical Microbiology 1998; 36(11)
:3239-42
• Madigan M, Martinko J, Parker J. Biology of Microorganisms. Ninth edition
• A, Bosy, G.V. Kulkarni, M. Rosenberg and C.A.G. McCulloch. Relationship of oral malodor to periodontitis: Evidence of independence in discrete subpopulations. Journal of Periodontology 1994; 65:37-46.
• Rosenberg Mel. The science of Bad Breath. Scientific American 2002; 72-79.
• Bruce P,Boches S, Galvin J. Bacterial Diversity in human subgingival plaque.
Journal of Bacteriology 2001;183(12): 3770-83.
• De Boever E, Uzeda M, Loesche W. Relationship between volatile sulfur compounds,Bana-Hydrolyzing bacteria and gingival health in patients with and without complaints of oral malodor. Journal of Clinical Dentistry 1994;
4(4):114-119
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