Little cigars and cigarillos harbor diverse bacterial communities that differ between the tobacco and the wrapper

Abstract

Despite their potential importance with regard to infectious and chronic diseases among tobacco users, microbial constituents of tobacco products lack characterization. Specifically, to our knowledge, there are no data describing the bacterial diversity of little cigars or cigarillos. To address this knowledge gap, we tested four brands of little cigars and cigarillos. Tobacco and wrapper subsamples (n = 132) were separately subjected to DNA extraction, followed by PCR amplification of the V3V4 hypervariable region of the 16S rRNA gene, and sequencing using Illumina HiSeq. Sequences were analyzed using QIIME and Phyloseq implemented in R. We identified 2,681 operational taxonomic units across all products. Significant differences in alpha and beta diversity were observed between Swisher Sweets and Cheyenne products. Alpha and beta diversity was also significantly different between tobacco and wrapper subsamples within the same product. Beta diversity analyses of only tobacco samples identified no significant differences in the bacterial microbiota of different lots of the same products; however, the microbiota in the wrapper differed significantly across lots for all brands. Overall, Firmicutes were found to dominate in the wrapper, whereas Proteobacteria were most abundant in the tobacco. At the genus level, Bacillus and Lactobacillus dominated in the wrappers, and Staphylococcus and Pseudomonas dominated in the tobacco. Our findings suggest that the bacterial microbiota of little cigars and cigarillos is diverse and differs significantly between the tobacco and the wrapper, and across brands. Future work is necessary to evaluate the potential public health implications of these findings.

The Bacterial Communities of Little Cigars and Cigarillos Are Dynamic Over Time and Varying Storage Conditions

“…The present study sought to generate a comprehensive longitudinal characterization of the bacterial microbiota present in little cigar products under different storage conditions, using a cultivation-independent next-generation sequencing approach. In a previous study, where the microbiotas of little cigar tobacco and wrapper components were characterized cross-sectionally at baseline for different brands and lots (Chattopadhyay et al, 2019), we showed that little cigar products have a composite microbiota, where tobacco and wrapper components harbor significantly different bacterial microbiotas. The little cigar tobacco and wrapper components were shown to be significantly different in their alpha- and beta- diversity, as well as in their bacterial microbiota composition, for all little cigar brands tested.…”

“…Detailed cross-sectional analysis of the baseline (day 0) microbiota associated with different brands, lots, and components of little cigar products was performed in our previous cross-sectional study (Chattopadhyay et al, 2019). Briefly, the heatmap displayed in Figure 1 summarizes the differences in bacterial community composition between the different brands of little cigars, and between tobacco and wrapper.…”

“…Several recent studies using next-generation sequencing have characterized the bacterial communities in a variety of tobacco products, including smokeless tobacco (Han et al, 2016; Tyx et al, 2016; Smyth et al, 2017), cigarettes (Chopyk et al, 2017a, b) and little cigars (Chattopadhyay et al, 2019). Our group recently showed that cigarette tobacco was dominated by bacteria from the genera Pseudomonas , Pantoea , and Bacillus , many of which contain potentially pathogenic species – such as Pseudomonas putida —that cigarette smokers may be exposed to over time (Chopyk et al, 2017a, b).…”

“…Smokeless tobacco has been shown to be dominated by bacteria from the Firmicutes , Proteobacteria , Actinobacteria , and Bacteroidetes phyla (Han et al, 2016; Tyx et al, 2016; Smyth et al, 2017). In an initial cross-sectional study focused on identifying the microbiota present in little cigars and cigarillos (Chattopadhyay et al, 2019), we have shown that little cigars are composite products in regards to their microbiota, with the tobacco being dominated by bacteria from the genera Pantoea, Pseudomonas and Staphylococcus , and the little cigar wrapper being dominated by bacteria from the Lactobacillus and Bacillus genera.…”

Microbiology of the American Smokeless Tobacco

“…Similarly, other groups have identified a vast diversity of bacterial species present in cigars, cigarettes, and smokeless tobacco products (Chattopadhyay et al 2019 ; Chopyk et al 2017a ; Sapkota et al 2010 ). These studies have shown that several genera, such as Bacillus , Pseudomonas , Pantoea , and Staphylococcus in particular, tend to be present in most or all tobacco products.…”