1887

Abstract

This study aimed to investigate the effect of smoking on the buccal microbiome and to analyse the descriptive ability of each of the seven hypervariable regions in their 16S rRNA genes.

Microbiome compositions of 40 buccal swab samples collected from smokers ( =20) and non-smokers ( =20) were determined using 16S rRNA sequencing. Seven different 16S rRNA hypervariable regions (V2, V3, V4, V6-7, V8 and V9) in each sample were amplified using the Ion Torrent 16S Metagenomics kit and were sequenced on the Ion S5 instrument.

Seven hypervariable regions in the 16S rRNA gene were successfully sequenced for all samples tested. The data obtained with the V2 region was found to be informative but the consensus data generated according to a number of operational taxonomic unit reads gathered from all seven hypervariable regions gave the most accurate result. At the phylum level, no statistically significant difference was found between smokers and non-smokers whereas relative abundances of , , , and showed significant increases in the smoker group (-adj=0.05). Alpha diversity results did not show a significant difference between the two groups; however, beta diversity analysis indicated that samples of smoker and non-smoker groups had a tendency to be clustered within themselves.

The results of the current study indicate that smoking is a factor influencing buccal microbiome composition. In addition, sequencing of all seven hypervariable regions yielded more accurate results than those with any of the single variable regions.

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2019-08-01
2020-01-28
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References

  1. Peterson J, Garges S, Giovanni M, McInnes P, Wang L et al. The NIH human microbiome project. Genome Res 2009;19:2317–2323
    [Google Scholar]
  2. Dewhirst FE, Chen T, Izard J, Paster BJ, Tanner AC et al. The human oral microbiome. J Bacteriol 2010;192:5002–5017 [CrossRef]
    [Google Scholar]
  3. Jia G, Zhi A, Lai PFH, Wang G, Xia Y et al. The oral microbiota – a mechanistic role for systemic diseases. Br Dent J 2018;224:447–455 [CrossRef]
    [Google Scholar]
  4. Bradshaw DJ, Marsh PD. Analysis of pH-driven disruption of oral microbial communities in vitro. Caries Res 1998;32:456–462 [CrossRef]
    [Google Scholar]
  5. Sachdeo A, Haffajee AD, Socransky SS. Biofilms in the edentulous oral cavity. J Prosthodont 2008;17:348–356 [CrossRef]
    [Google Scholar]
  6. Sampaio-Maia B, Caldas IM, Pereira ML, Perez-Mongiovi D, Araujo R. The oral microbiome in health and its implication in oral and systemic diseases. Adv Appl Microbiol 2016;97:171–210
    [Google Scholar]
  7. Papaioannou W, Gizani S, Haffajee AD, Quirynen M, Mamai-Homata E et al. The microbiota on different oral surfaces in healthy children. Oral Microbiol Immunol 2009;24:183–189 [CrossRef]
    [Google Scholar]
  8. Marsh PD, Martin MV, Lewis MAO, Williams D. Oral Microbiology, 5th ed. Edinburgh: Elsevier; 2009
    [Google Scholar]
  9. Mattila KJ, Pussinen PJ, Paju S. Dental infections and cardiovascular diseases: a review. J Periodontol 2005;76:2085–2088 [CrossRef]
    [Google Scholar]
  10. Wu J, Peters BA, Dominianni C, Zhang Y, Pei Z et al. Cigarette smoking and the oral microbiome in a large study of American adults. ISME J 2016;10:2435–2446 [CrossRef]
    [Google Scholar]
  11. Mason MR, Preshaw PM, Nagaraja HN, Dabdoub SM, Rahman A et al. The subgingival microbiome of clinically healthy current and never smokers. ISME J 2015;9:268–272 [CrossRef]
    [Google Scholar]
  12. Sapkota AR, Berger S, Vogel TM. Human pathogens abundant in the bacterial metagenome of cigarettes. Environ Health Perspect 2010;118:351–356 [CrossRef]
    [Google Scholar]
  13. Kanwar A, Sah K, Grover N, Chandra S, Singh R. Long-term effect of tobacco on resting whole mouth salivary flow rate and pH: an institutional based comparative study. European J Gen Dent 2013;2:296–299
    [Google Scholar]
  14. 16S metagenomic sequencing library preparation-preparing 16S ribosomal RNA gene amplicons for the Illumina MiSeq system. https://support.illumina.com/documents/documentation/chemistry_documentation/16s/16s-metagenomic-library-prep-guide-15044223-b.pdf
  15. Chakravorty S, Helb D, Burday M, Connell N, Alland D. A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria. J Microbiol Meth 2007;69:330–339 [CrossRef]
    [Google Scholar]
  16. Graspeuntner S, Loeper N, Künzel S, Baines JF, Rupp J. Selection of validated hypervariable regions is crucial in 16S-based microbiota studies of the female genital tract. Sci Rep 2018;8:9678 [CrossRef]
    [Google Scholar]
  17. Thermo scientific GeneJET genomic DNA purification kit. 2016;https://www.thermofisher.com/order/catalog/product/K0721
  18. Chaitali Parikh EB, Stephen J. Rapid 16S next generation sequencing for bacterial identification in polymicrobial samples: Thermo Fisher Scientific. 2018;https://www.slideshare.net/ThermoFisher/c-parikh-asm2015posterfinal
  19. Ion Reporter™ 5.0 software: Thermo Fisher Scientific. https://assets.thermofisher.com/TFS-Assets/LSG/manuals/IonReporter_ v50_Help.pdf
  20. Saeb ATM, Al-Rubeaan KA, Aldosary K, Udaya Raja GK, Mani B et al. Relative reduction of biological and phylogenetic diversity of the oral microbiota of diabetes and pre-diabetes patients. Microb Pathog 2019;128:215–229 [CrossRef]
    [Google Scholar]
  21. Zhang J, Sun QL, Zeng ZG, Chen S, Sun L. Microbial diversity in the deep-sea sediments of Iheya North and Iheya ridge, Okinawa Trough. Microbiol Res 2015;177:43–52 [CrossRef]
    [Google Scholar]
  22. Zarco MF, Vess TJ, Ginsburg GS. The oral microbiome in health and disease and the potential impact on personalized dental medicine. Oral Dis 2012;18:109–120 [CrossRef]
    [Google Scholar]
  23. Lazarevic V, Whiteson K, Hernandez D, Francois P, Schrenzel J. Study of inter- and intra-individual variations in the salivary microbiota. BMC Genomics 2010;11:523 [CrossRef]
    [Google Scholar]
  24. Chao A. Non-parametric estimation of the classes in a population. Scand J Stat 1984;11:265–270
    [Google Scholar]
  25. Morris EK, Caruso T, Buscot F, Fischer M, Hancock C et al. Choosing and using diversity indices: insights for ecological applications from the German biodiversity exploratories. Ecol Evol 2014;4:3514–3524 [CrossRef]
    [Google Scholar]
  26. Doel JJ, Benjamin N, Hector MP, Rogers M, Allaker RP. Evaluation of bacterial nitrate reduction in the human oral cavity. Eur J Oral Sci 2005;113:14–19 [CrossRef]
    [Google Scholar]
  27. Liddle L, Burleigh MC, Monaghan C, Muggeridge DJ, Sculthorpe N et al. Variability in nitrate-reducing oral bacteria and nitric oxide metabolites in biological fluids following dietary nitrate administration: an assessment of the critical difference. Nitric Oxide 2019;83:1–10 [CrossRef]
    [Google Scholar]
  28. Karasneh JA, Al Habashneh RA, Marzouka NAS, Thornhill MH. Effect of cigarette smoking on subgingival bacteria in healthy subjects and patients with chronic periodontitis. BMC Oral Health 2017;17:64 [CrossRef]
    [Google Scholar]
  29. Kato I, Vasquez AA, Moyerbrailean G, Land S, Sun J et al. Oral microbiome and history of smoking and colorectal cancer. J Epidemiol Res 2016;2:92–101 [CrossRef]
    [Google Scholar]
  30. Washio J, Sato T, Koseki T, Takahashi N. Hydrogen sulfide-producing bacteria in tongue biofilm and their relationship with oral malodour. J Med Microbiol 2005;54:889–895 [CrossRef]
    [Google Scholar]
  31. Crielaard W, Zaura E, Schuller AA, Huse SM, Montijn RC et al. Exploring the oral microbiota of children at various developmental stages of their dentition in the relation to their oral health. BMC Med Genomics 2011;4:22 [CrossRef]
    [Google Scholar]
  32. Tanner AC, Kent RL Jr, Holgerson PL, Hughes CV, Loo CY et al. Microbiota of severe early childhood caries before and after therapy. J Dent Res 2011;90:1298–1305 [CrossRef]
    [Google Scholar]
  33. Kato I, Vasquez A, Moyerbrailean G, Land S, Djuric Z et al. Nutritional correlates of human oral microbiome. J Am Coll Nutr 2017;36:88–98 [CrossRef]
    [Google Scholar]
  34. Belstrøm D, Holmstrup P, Nielsen CH, Kirkby N, Twetman S et al. Bacterial profiles of saliva in relation to diet, lifestyle factors, and socioeconomic status. J Oral Microbiol 2014;6:23609 [CrossRef]
    [Google Scholar]
  35. Yamashita Y, Takeshita T. The oral microbiome and human health. J Oral Sci 2017;59:201–206 [CrossRef]
    [Google Scholar]
  36. Belstrøm D, Holmstrup P, Bardow A, Kokaras A, Fiehn NE et al. Temporal stability of the salivary microbiota in oral health. Plos One 2016;11:e0147472–e [CrossRef]
    [Google Scholar]
  37. Utter DR, Mark Welch JL, Borisy GG. Individuality, stability, and variability of the plaque microbiome. Front Microbiol 2016;7:564 [CrossRef]
    [Google Scholar]
  38. Rasiah IA, Wong L, Anderson SA, Sissons CH. Variation in bacterial DGGE patterns from human saliva: over time, between individuals and in corresponding dental plaque microcosms. Arch Oral Biol 2005;50:779–787 [CrossRef]
    [Google Scholar]
  39. Zaura E, Nicu EA, Krom BP, Keijser BJF. Acquiring and maintaining a normal oral microbiome: current perspective. Front Cell Infect Microbiol 2014;4:85 [CrossRef]
    [Google Scholar]
  40. Moon JH, Lee JH, Lee JY. Subgingival microbiome in smokers and non-smokers in Korean chronic periodontitis patients. Mol Oral Microbiol 2015;30:227–241 [CrossRef]
    [Google Scholar]
  41. Brook I. The impact of smoking on oral and nasopharyngeal bacterial flora. J Dent Res 2011;90:704–710 [CrossRef]
    [Google Scholar]
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