Abstract
BackgroundThere is a paucity of data regarding the microbial constituents of tobacco products and their impacts on public health. Moreover, there has been no comparative characterization performed on the bacterial microbiota associated with the addition of menthol, an additive that has been used by tobacco manufacturers for nearly a century. To address this knowledge gap, we conducted bacterial community profiling on tobacco from user- and custom-mentholated/non-mentholated cigarette pairs, as well as a commercially-mentholated product. Total genomic DNA was extracted using a multi-step enzymatic and mechanical lysis protocol followed by PCR amplification of the V3-V4 hypervariable regions of the 16S rRNA gene from five cigarette products (18 cigarettes per product for a total of 90 samples): Camel Crush, user-mentholated Camel Crush, Camel Kings, custom-mentholated Camel Kings, and Newport Menthols. Sequencing was performed on the Illumina MiSeq platform and sequences were processed using the Quantitative Insights Into Microbial Ecology (QIIME) software package.ResultsIn all products, Pseudomonas was the most abundant genera and included Pseudomonas oryzihabitans and Pseudomonas putida, regardless of mentholation status. However, further comparative analysis of the five products revealed significant differences in the bacterial compositions across products. Bacterial community richness was higher among non-mentholated products compared to those that were mentholated, particularly those that were custom-mentholated. In addition, mentholation appeared to be correlated with a reduction in potential human bacterial pathogens and an increase in bacterial species resistant to harsh environmental conditions.ConclusionsTaken together, these data provide preliminary evidence that the mentholation of commercially available cigarettes can impact the bacterial community of these products.
Highlights
There is a paucity of data regarding the microbial constituents of tobacco products and their impacts on public health
Reynolds Tobacco Co., Winston-Salem, NC, USA), where the capsule within the filter was subsequently not crushed during the study; (2) Camel Crush, regular, fresh (CCM), where the capsule was subsequently crushed during the study to release a menthol-containing solution into the cigarette filter (CCM); and (3) a commercially mentholated brand, Newport Menthol Box (NMB) (Lorillard Tobacco Co., Greensboro, NC, USA)
Members from the Pseudomonas genus were comprised of 15 unique Operational taxonomic unit (OTU), with 7 Pseudomonas OTUs shared between all mentholation states (OTU#s 1532, 10, 134, 1868, 1886, 8, and 3)
Summary
There is a paucity of data regarding the microbial constituents of tobacco products and their impacts on public health. There has been no comparative characterization performed on the bacterial microbiota associated with the addition of menthol, an additive that has been used by tobacco manufacturers for nearly a century. To address this knowledge gap, we conducted bacterial community profiling on tobacco from user- and custom-mentholated/non-mentholated cigarette pairs, as well as a commercially-mentholated product. Non-menthol cigarettes do contain low levels of menthol, levels in cigarette products labeled as mentholated are 50–5000 times higher [7]. Several brands of cigarettes (e.g., Camel Crush) have capsules embedded in the filter, which can be “crushed” by the user to release a menthol-containing solution. Young adults, minority groups, adult women, and members of low-income households are the primary consumers of menthol cigarettes [2, 9, 10]
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