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Journal of Published by the American Society for Microbiology Clinical Microbiology
Copyright 2003, American Society for Microbiology
J Clin Microbial. 2003 February; 41 (2):558－563
Diversity of Bacterial Populations on the Tongue Dorsa of Patients with Halitosis and Healthy Patients
C.E.Kazor, P.M.Mitchell, A.M.Lee, L.N.Stokes, W.J.Loesche, F.E.Dewhirst, and B.J.Paster
|Department of Microbiology and Immunology, School of Medicine, University of Michigan , Ann Arbor , Michigan
Department of Molecular Genetics, The Forsyth Institute
Department of Oral and Developmental Biology, Harvard School of Dental
Medecine.Boston.MassachusettsReceived August 5, 2002 ; Revised November
9, 2002 ; Accepted November 21, 2002 .
The primary purpose of the present study was
to compare the microbial profiles of the tongue dorsa of healthy
subjects and subjects with halitosis by using culture-independent
molecular methods. Our overall goal was to determine the bacterial
diversity on the surface of the tongue dorsum as part of our ongoing
efforts to identify all cultivable and not-yet-cultivated species of the
oral cavity. Tongue dorsum scrapings were analyzed from healthy
subjects with no complaints of halitosis and subjects with halitosis,
defined as an organoleptic score of 2 or more and volatile sulfur
compound levels greater than 200 ppb. 16S rRNA genes from DNA isolated
from tongue dorsum scrapings were amplified by PCR with universally
conserved bacterial primers and cloned into Escherichia coli. Typically,
50 to 100 clones were analyzed from each subject. Fifty-one strains
isolated from the tongue dorsa of healthy subjects were also analyzed.
Partial sequences of approximately 500 bases of cloned inserts from the
16S rRNA genes of isolates were compared with sequences of known species
or phylotypes to determine species identity or closest relatives.
Nearly complete sequences of about 1,500 bases were obtained for
potentially novel species or phylotypes. In an analysis of approximately
750 clones, 92 different bacterial species were identified. About half
of the clones were identified as phylotypes, of which 29 were novel to
the tongue microbiota. Fifty-one of the 92 species or phylotypes were
detected in more than one subject. Those species most associated with
healthy subjects were Streptococcus salivarius , Rothia mucilaginosa , and an uncharacterized species of Eubacterium (strain FTB41). Streptococcus salivarius
was the predominant species in healthy subjects,as it represented 12 to
40% of the total clones analyzed from each healthy subject. Overall,
the predominant microbiota on the tongue dorsa of healthy subjects was
different from that on the tongue dorsa of subjects with halitosis.
Those species most associated with halitosis were Atopobium parvulum, a phylotype(clone BS095) of Dialister, Eubacterium sulci, a phylotype(clone DR034) of the uncultivated phylum TM7, Solobacterium moorei , and a phylotype(clone BW009) of Streptcoccus.
On the basis of our ongoing efforts to obtain full 16S rRNA sequences
for all cultivable and not-yet-cultivated species that colonize the oral
cavity, there are now over 600 species.