Abstract
Smokeless tobacco products contain numerous chemical compounds, including known human carcinogens. Other smokeless tobacco constituents, including bacteria, may also contribute to adverse health effects among smokeless tobacco users. However, there is a lack of data regarding the microbial constituents of smokeless tobacco. Our goal was to characterize the bacterial microbiota of different smokeless tobacco products and evaluate differences across product types and brands. DNA was extracted from 15 brands of smokeless tobacco products (including dry snuff, moist snuff, snus, and Swedish snus) and 6 handmade products (e.g., toombak) using an enzymatic and mechanical lysis approach. Bacterial community profiling was performed using PCR amplification of the V1–V2 hypervariable region of the 16S rRNA gene, followed by 454 pyrosequencing of the resulting amplicons and sequence analysis using the QIIME package. Total viable counts were also determined to estimate the number of viable bacteria present in each product. Average total viable counts ranged from 0 to 9.35 × 107 CFU g−1. Analysis of the 16S rRNA gene sequences revealed high bacterial diversity across the majority of products tested: dry snuff products where characterized by the highest diversity indices compared to other products. The most dominant bacterial phyla across all products were Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. Significant differences in both bacterial community composition and in silico predicted gene content were observed between smokeless tobacco product types and between brands of specific smokeless tobacco products. These data are useful in order to comprehensively address potential health risks associated with the use of smokeless tobacco products.
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Acknowledgements
We would like to thank Ghazi Zaatari, M.D. (Department of Pathology and Laboratory Medicine, American University of Beirut; Beirut, Lebanon) for providing the toombak samples and alkalinizing agent and Dr. Angel J. Rivera for his helpful review and input on this manuscript.
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This study was funded by a contract (200-2012-M-51035) from the Centers for Disease Control and Prevention (CDC) and by grant number P50CA180523 from the National Cancer Institute and FDA Center for Tobacco Products (CTP) awarded to the University of Maryland. The content is solely the responsibility of the authors and does not necessarily represent the official views of CDC, NIH, or the Food and Drug Administration.
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Smyth, E.M., Kulkarni, P., Claye, E. et al. Smokeless tobacco products harbor diverse bacterial microbiota that differ across products and brands. Appl Microbiol Biotechnol 101, 5391–5403 (2017). https://doi.org/10.1007/s00253-017-8282-9
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DOI: https://doi.org/10.1007/s00253-017-8282-9