Purpose: The purpose of the present study was to develop a novel selective medium (OVSM) and useful identification method for oral Veillonella species, to assess the prevalence of this species using the selective medium, and to investigate whether the monitoring of oral Veillonella species levels is useful as a clinical indicator for the diagnosis of halitosis. Methods: To examine bacterial populations in the oral cavity, a novel selective medium (OVSM) and multiplex PCR method were developed for isolating and identifying oral Veillonella species. OVSM consists of tryptic soy agar, sheep blood, sodium lactate, vancomycin, and oxacillin. Results: Oral Veillonella species grew well on OVSM. Oral Veillonella species accounted for 5.4% and 27.2%, respectively, of all bacteria in the tongue biofilm samples of healthy and halitosis subjects. Oral Veillonella species produced larger amounts of volatile sulfur compounds than other representative oral bacteria. Conclusion: These results indicate that monitoring the levels of these organisms is useful as a clinical indicator for the diagnosis of halitosis.
References
[1]
Marchandin, H., Teyssier, C., Campos, J., Jean-Pierre, H., Roger, F., Gay, B., Carlier, J.P. and Jumas-Bilak, E. (2010) Negativicoccus succinicivorans Gen. Nov., Sp. Nov., Isolated from Human Clinical Samples, Emended Description of the Family Veillonellaceae and Description of Negativicutes Classis Nov., Selenomonadales Ord. Nov. and Acidaminococcaceae Fam. Nov. in the Bacterial Phylum Firmicutes. International Journal of Systematic and Evolutionary Microbiology, 60, 1271-1279. https://doi.org/10.1099/ijs.0.013102-0
[2]
Mays, T.D., Holdeman, L.V., Moore, W.E.C., Rogosa, M. and Johnson, J.L. (1982) Taxonomy of the Genus Veillonella Prévot. International Journal of Systematic and Evolutionary Microbiology, 32, 28-36. https://doi.org/10.1099/00207713-32-1-28
[3]
Foubert, E.L. and Douglas, H.C. (1948) Studies on the Anaerobic Micrococci. II. The Fermentation of Lactate by Micrococcus lactilyticus. Journal of Bacteriology, 56, 35-36. https://doi.org/10.1128/jb.56.1.35-36.1948
[4]
Ng, S.K. and Hamilton, I.R. (1971) Lactate Metabolism by Veillonella parvula. Journal of Bacteriology, 105, 999-1005. https://doi.org/10.1128/jb.105.3.999-1005.1971
[5]
Mashima, I., Theodorea, C.F., Thaweboon, B., Thaweboon, S., Scannapieco, F.A. and Nakazawa, F. (2017) Exploring the Salivary Microbiome of Children Stratified by the Oral Hygiene Index. PLOS ONE, 12, e01852. https://doi.org/10.1371/journal.pone.0185274
[6]
Mashima, I., Theodorea, C.F., Thaweboon, B., Thaweboon, S., Vichayanrat, T., Scannapieco, F.A. and Nakazawa, F. (2019) Characterization of Salivary Microbiome in Healthy Thai Children. Asian Pacific Journal of Tropical Medicine, 12, 163-169. https://doi.org/10.4103/1995-7645.257116
[7]
Marriott, D., Stark, D. and Harkness, J. (2007) Veillonella parvula Discitis and Secondary Bacteremia: A Rare Infection Complicating Endoscopy and Colonoscopy? Journal of Clinical Microbiology, 45, 672-674. https://doi.org/10.1128/JCM.01633-06
[8]
Mashima, I., Miyoshi-Akiyama, T., Tomida, J., Kutsuna, R., Washio, J., Takahashi, N., Nakazawa, F., Sato, T. and Kawamura, Y. (2019) Draft Genome Sequences of Two Veillonella tobetsuensis Clinical Isolates from Intraoperative Bronchial Fluids of Elderly Patients with Pulmonary Carcinoma. Microbiology Resource Announcements, 8, e00397-19. https://doi.org/10.1128/MRA.00397-19
[9]
Periasamy, S. and Kolenbrander, P.E. (2010) Central Role of the Early Colonizer Veillonella Sp. in Establishing Multispecies Biofilm Communities with Initial, Middle, and Late Colonizers of Enamel. Journal of Bacteriology, 192, 2965-2972. https://doi.org/10.1128/JB.01631-09
[10]
Kanasi, E., Dewhirst, F.E., Chalmers, N.I., Kent, R., Moore, A., Hughes, C.V., Pradhan, N., Loo, C.Y. and Tanner, C.R. (2010) Clonal Analysis of the Microbiota of Severe Early Childhood Caries. Caries Research, 44, 485-497. https://doi.org/10.1159/000320158
[11]
Khemaleelakul, S., Baumgartner, J.C. and Pruksakorn, S. (2009) Identification of Bacteria in Acute Endodontic Infections and Their Antimicrobial Susceptibility. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology, 94, 746-755. https://doi.org/10.1067/moe.2002.129535
[12]
Baumgartener, J.C. and Falkler, W.A. (1991) Bacteria in the Apical 5 mm of Infected Root Canals. Journal of Endodontics, 17, 380-383. https://doi.org/10.1016/S0099-2399(06)81989-8
[13]
Peters, L.B., Wesselink, P.R., Buijs, J.F. and van Winkelhoff, A.J. (2001) Viable Bacteria in Root Dentinal Tubules of Teeth with Apical Periodontitis. Journal of Endodontics, 27, 76-81. https://doi.org/10.1097/00004770-200102000-00002
[14]
Heller, D., Silva-Boghossian, C.M., do Souto, R.M. and Colombo, A.P. (2012) Subgingival Microbial Profiles of Generalized Aggressive and Chronic Periodontal Diseases. Archives of Oral Biology, 57, 973-980. https://doi.org/10.1016/j.archoralbio.2012.02.003
[15]
Rogosa, M. (1965) The Genus Veillonella IV. Serological Groupings, and Genus and Species Emendations. Journal of Bacteriology, 90, 704-709. https://doi.org/10.1128/jb.90.3.704-709.1965
[16]
Byun, R., Carlier, J.P., Jacques, N.A., Marchandin, H. and Hunter, N. (2007) Veillonella denticariosi Sp. Nov., Isolated from Human Carious Dentine. International Journal of Systematic and Evolutionary Microbiology, 57, 2844-2848. https://doi.org/10.1099/ijs.0.65096-0
[17]
Arif, N., Do, T., Byun, R., Sheehy, E., Clark, D., Gilbert, S.C. and Beighton, D. (2008) Veillonella rogosae Sp. Nov., an Anaerobic, Gram-Negative Coccus Isolated from Dental Plaque. International Journal of Systematic and Evolutionary Microbiology, 58, 581-584. https://doi.org/10.1099/ijs.0.65093-0
[18]
Mashima, I., Kamaguchi, A., Miyakawa, H. and Nakazawa, F. (2013) Veillonella tobetsuensis Sp. Nov., an Anaerobic, Gram-Negative Coccus Isolated from Human Tongue Biofilms. International Journal of Systematic and Evolutionary Microbiology, 63, 1443-1449. https://doi.org/10.1099/ijs.0.042515-0
[19]
Mashima, I., Liao, Y.C., Miyakawa, H., Theodorea, C.F., Thawboon, B., Thawboon, S., Scannapieco, F.A. and Nakazawa, F. (2018) Veillonella infantium Sp. Nov., an Anaerobic, Gram-Stain-Negative Coccus Isolated from Tongue Biofilm of a Thai Child. International Journal of Systematic and Evolutionary Microbiology, 68, 1101-1106. https://doi.org/10.1099/ijsem.0.002632
[20]
Kazor, C.E., Mitchell, P.M., Lee, A.M., Stokes, L.N., Loesche, W.J., Dewhirst, F.E. and Paster, B.J. (2003) Diversity of Bacterial Populations on the Tongue Dorsa of Patients with Halitosis and Healthy Patients. Journal of Clinical Microbiology, 41, 558-563. https://doi.org/10.1128/JCM.41.2.558-563.2003
[21]
Persson, S., Edlund, M.B., Claesson, R. and Carlsson, J. (1990) The Formation of Hydrogen Sulfide and Methyl Mercaptan by Oral Bacteria. Oral Microbiology and Immunology, 5, 195-201. https://doi.org/10.1111/j.1399-302X.1990.tb00645.x
[22]
Yaegaki, K. and Sanada, K. (1992) Biochemical and Clinical Factors Influencing Oral Malodor in Periodontal Patients. Journal of Periodontology, 63, 783-789. https://doi.org/10.1902/jop.1992.63.9.783
[23]
Faveri, M., Feres, M., Shibli, J.A., Hayacibara, R.F., Hayacibara, M.M. and de Figueiredo, L.C. (2006) Microbiota of the Dorsum of the Tongue after Plaque Accumulation; an Experimental Study in Humans. Journal of Periodontology, 77, 1539-1546. https://doi.org/10.1902/jop.2006.050366
[24]
Fukamachi, H., Nakano, Y., Okano, S., Shibata, Y., Abiko, Y. and Yamashita, Y. (2005) High Production of Methyl Mercaptan by L-Methionine-Deamino-Mercaptomethane Lyase from Treponema denticola. Biochemical and Biophysical Research Communications, 331, 127-131. https://doi.org/10.1016/j.bbrc.2005.03.139
[25]
Washio, J., Sato, T., Koseki, T. and Takahashi, N. (2005) Hydrogen Sulfide-Producing Bacteria in Tongue Biofilm and Their Relationship with Oral Malodour. Journal of Medical Microbiology, 54, 889-895. https://doi.org/10.1099/jmm.0.46118-0
[26]
Beighton, D., Clark, D., Hanakuka, B., Gilbert, S. and Do, T. (2008) The Predominant Cultivable Veillonella Spp. of the Tongue of Healthy Adults Identified Using rpoB Sequencing. Oral microbiology Immunology, 23, 344-347. https://doi.org/10.1111/j.1399-302X.2007.00424.x
[27]
Mashima, I., Theodorea, C.F., Thaweboon, B., Thaweboon, S. and Nakazawa, F. (2016) Identification of Veillonella Species in the Tongue Biofilm by Using a Novel One-Step Polymerase Chain Reaction Method. PLOS ONE, 11, e0157516. https://doi.org/10.1371/journal.pone.0157516
[28]
Djais, A.A., Theodorea, C.F., Mashima, I., Otomo, M., Saitoh, M. and Nakazawa, F. (2019) Identification and Phylogenetic Analysis of Oral Veillonella Species Isolated from the Saliva of Japanese Children. F1000Research, 8, 616. https://doi.org/10.12688/f1000research.18506.5
[29]
Rogosa, M., Fitzgerald, R.J., Mackintosh, M.E. and Beaman, J. (1958) Improved Medium for Selective Isolation of Veillonella. Journal of Bacteriology, 76, 455-456. https://doi.org/10.1128/jb.76.4.455-456.1958
[30]
Marchandin, H., Teyssier, C., Siméon, D.B., Jean, P.H., Carriere, C. and Jumas, B.E. (2003) Intrachromosomal Heterogeneity between the Four 16S rRNA Gene Copies in the Genus Veillonella: Implications or Phylogeny and Taxonomy. Microbiology, 149, 1493-1501. https://doi.org/10.1099/mic.0.26132-0
[31]
Tsuzukibashi, O., Uchibori, S., Kobayashi, T., Umezawa, K., Mashimo, C., Nambu, T., Saito, M., Hashizume-Takizawa, T. and Ochiai, T. (2017) Isolation and Identification Methods of Rothia Species in Oral Cavities. Journal of Microbiological Methods, 134, 21-26. https://doi.org/10.1016/j.mimet.2017.01.005
[32]
Krespi, Y.P., Shrime, M.G. and Kacker, A. (2006) The Relationship between Oral Malodor and Volatile Sulfur Compound-Producing Bacteria. Otolaryngology, 135, 671-676. https://doi.org/10.1016/j.otohns.2005.09.036
[33]
Hughes, F.J. and McNab, R. (2010) Oral Malodour—A Review. Archives of Oral Biology, 53, S1-S7. https://doi.org/10.1016/S0003-9969(08)70002-5
[34]
Morita, M. and Wang, H.L. (2001) Association between Oral Malodor and Adult Periodontitis: A Review. Journal of Clinical Periodontology, 28, 813-819. https://doi.org/10.1034/j.1600-051x.2001.028009813.x
[35]
Hiranmayi, K.V., Sirisha, K., Ramoji, R.M.V. and Sudhakar, P. (2017) Novel Pathogens in Periodontal Microbiology. Journal of Pharmacy and Bioallied Sciences, 9, 155-163. https://doi.org/10.4103/jpbs.JPBS_288_16
