Characterization of Bacterial Communities in Selected Smokeless Tobacco Products Using 16S rDNA Analysis

PLoS One. 2016 Jan 19;11(1):e0146939. doi: 10.1371/journal.pone.0146939. eCollection 2016.

Abstract

The bacterial communities present in smokeless tobacco (ST) products have not previously reported. In this study, we used Next Generation Sequencing to study the bacteria present in U.S.-made dry snuff, moist snuff and Sudanese toombak. Sample diversity and taxonomic abundances were investigated in these products. A total of 33 bacterial families from four phyla, Actinobacteria, Firmicutes, Proteobacteria and Bacteroidetes, were identified. U.S.-produced dry snuff products contained a diverse distribution of all four phyla. Moist snuff products were dominated by Firmicutes. Toombak samples contained mainly Actinobacteria and Firmicutes (Aerococcaceae, Enterococcaceae, and Staphylococcaceae). The program PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) was used to impute the prevalence of genes encoding selected bacterial toxins, antibiotic resistance genes and other pro-inflammatory molecules. PICRUSt also predicted the presence of specific nitrate reductase genes, whose products can contribute to the formation of carcinogenic nitrosamines. Characterization of microbial community abundances and their associated genomes gives us an indication of the presence or absence of pathways of interest and can be used as a foundation for further investigation into the unique microbiological and chemical environments of smokeless tobacco products.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteria / classification*
  • Bacteria / genetics
  • Bacterial Toxins / genetics
  • DNA, Bacterial / analysis*
  • DNA, Ribosomal / analysis*
  • Drug Resistance, Bacterial
  • High-Throughput Nucleotide Sequencing
  • Metagenome
  • Phylogeny
  • RNA, Ribosomal, 16S / analysis*
  • Software
  • Tobacco, Smokeless / classification
  • Tobacco, Smokeless / microbiology*
  • United States

Substances

  • Bacterial Toxins
  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S

Grants and funding

Battelle Analytical Services, through a contract with U.S. Centers for Disease control and Prevention, provided support in the form of salaries for authors (LMK), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.