Halitosis-Associated Bacteria
3455 Bacterial Culture and Broad Range PCR for Halitosis-Associated
Bacteria
Objectives: Oral microorganisms are responsible for a large proportion of the 50-80 million cases of halitosis (oral malodor) Americans experience at some point in their lives. While some of the microorganisms associated with halitosis such as the volatile sulfur compound-producing bacteria, have been identified, the microbiology of halitosis is not well characterized.
This study compared bacterial culture and the broad-range polymerase chain reaction (PCR) in identifying cultivable and non-cultivable
Microorganisms associated with oral malodor. Methods: Samples were obtained from 6 adult subjects with persistent halitosis defined by organoleptic assessment (>2), elevated (>250 ppb) halimeter reading, and by a high Halitox test.
Bacterial samples obtained from the tongue were dispersed in sterile buffer, distributed onto enriched tryptic soy agar and anaerobically incubated at 35oC for 5-7 days. One hundred randomly selected colonies were subcultured and speciated from each sample. 16S rDNA was also amplified by PCR from aliquots of the original sample using primers to highly conserved regions.
The PCR product was cloned in E. coli and 100 randomly selected clones
From each sample were sequenced and identified. Results: Bacterial culture of tongue samples from subjects with halitosis demonstrated (in decreasing order) significant proportions of veillonella, actinomyces,
megasphaera, Streptococcus parasanguinis, Campylobacter concisus, and neisseria as well as unspeciated bacteria. Broad range PCR of the same samples revealed similar species as well as a diverse array of uncultured bacteria including species of atopobium, erysipelothrix, firmicutes, granulicatella, porphyromonas, and prevotella as well as Solobacterium moorei, this later species having recently been implicated in oral malodor.
Conclusions:
Broad range PCR identifies a broader group of halitosis-related bacteria than does traditional bacterial culture including rarely cultured, uncultured, and some, as yet, unidentified bacteria.
M.M. ZAMBON, D. GERBER, V.I. HARASZTHY, and J.J. ZAMBON, State
University of New York - Buffalo, USA