Mentholation triggers brand-specific shifts in the bacterial microbiota of commercial cigarette products

et al. 2021

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Previous studies have characterized bacterial communities in menthol versus non-menthol cigarettes. However, these studies evaluated commercial cigarettes, for which levels of chemical constituents are largely unknown, and therefore, could not assess the impact of varying nicotine and menthol concentrations on tobacco bacterial communities. To address this knowledge gap, we performed time-series experiments using SPECTRUM research cigarettes with varying nicotine and menthol levels. Cigarettes were incubated under three storage conditions for 14 days. Cigarette tobacco was then sub-sampled (n = 288), DNA extracted, and subjected to PCR amplification of the V3V4 region of the 16S rRNA gene, followed by Illumina HiSeq sequencing. Sequences were analyzed using QIIME and R. Incubation under varying conditions did not affect bacterial diversity. However, significant differences in bacterial communities were observed across varying nicotine concentrations in menthol and non-menthol products. For example, Pseudomonas spp. was negatively correlated with nicotine concentrations in menthol cigarettes. A significantly higher relative abundance of P. veronii and P. viridiflava was observed in menthols versus nonmenthols, while a significantly higher relative abundance of Bacillus foraminis and B. coagulans was found in non-menthols versus menthols. Additional bacteria (e.g., Staphylococcus spp., Jeotgalicoccus psychrophilus, and B. flexus) significantly changed in relative abundance between days 0 and 14. Our findings demonstrate that nicotine and menthol levels have a significant impact on the relative abundance of potential bacterial pathogens present in cigarettes. Future work is needed to demonstrate whether these tobacco-associated bacteria could be transferred to users while smoking, ultimately contributing to adverse respiratory impacts. Key points• Varying nicotine levels changes bacterial composition of research cigarettes.• Mentholation affects the tobacco bacterial microbiome.• SPECTRUM research cigarettes are dominated by Pseudomonas and Bacillus.

Section: Discussionsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Nicotine concentration and mentholation affect bacterial community diversity in SPECTRUM research cigarettes

et al. 2021

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“…Beyond human and plant bacterial pathogens, we also observed bacterial species that can tolerate heavy metals such as arsenic and degrade polycyclic hydrocarbons, such as nicotine and toluene, which are present in mainstream smoke. Similar bacterial genera were also previously observed in commercial cigarettes studied by our group (Malayil et al 2020 ). For example, B. flexus and P. veronii , which were observed in both hookah brands (Figs.…”

Section: Discussionsupporting

confidence: 90%

“…Total DNA was extracted under sterile laboratory conditions from 0.2 g of all hookah tobacco samples using previously published methods (Chopyk et al 2017a(Chopyk et al , 2017bChattopadhyay et al 2019;Smyth et al 2019;Malayil et al 2020). Additionally, negative extraction controls were included at every step of sample processing to ensure no exogenous DNA contaminated the samples.…”

Section: Dna Extraction 16s Rrna Gene Pcr Amplification and Sequencingmentioning

confidence: 99%

“…Nevertheless, existing studies on the negative health effects associated with hookahs have mostly focused on microbial loads in the various physical components of the water pipe. However, the microbial communities that may be present in the hookah tobacco itself have not been fully characterized, even though multiple studies have identified and characterized a plethora of microorganisms in other tobacco products (e.g., cigarettes, little cigars, cigarillos, and smokeless tobacco), including Actinomycetes , Acinetobacter , Bacillus , Burkholderia , Clostridium , Klebsiella , Pseudomonas , Serratia , Campylobacter , Enterococcus , Proteus , and Staphylococcus (Sapkota et al 2009 ; Tyx et al 2016 ; Han et al 2016 ; Chopyk et al 2017a , 2017b ; Smyth et al 2017 , 2019 ; Chattopadhyay et al 2019 ; Malayil et al 2020 ). To address this knowledge gap, we performed a time series experiment to characterize the bacterial communities present in hookah tobacco from two top brands: Al Fakher and Fumari.…”

Section: Introductionmentioning

confidence: 99%

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Bacterial communities of hookah tobacco products are diverse and differ across brands and flavors

et al. 2022

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Young adults are increasingly using non-cigarette products, such as hookahs, since they are perceived as healthier alternatives to cigarette smoking. However, hookah users are exposed to not only carcinogenic compounds but also microorganisms that may play an active role in the development of both infectious and chronic diseases among users. Nevertheless, existing hookah research in this area has focused only on microorganisms that may be transferred to users through the smoking apparatus and not on bacterial communities associated with hookah tobacco. To address this knowledge gap, we conducted time-series experiments on commercially available hookah brands (Al Fakher (flavors: two apple, mint, and watermelon) and Fumari (flavors: white gummy bear, ambrosia, and mint chocolate chill)) stored under three different temperature and relative humidity conditions over 14 days. To characterize bacterial communities, the total DNA was extracted on days 0, 5, 9, and 14, PCR-amplified for the V3V4 region of the bacterial 16S rRNA gene, sequenced on the Illumina HiSeq platform, and analyzed using R. Diversity (alpha and beta) analyses revealed that the microbiotas of Fumari and Al Fakher products differed significantly and that flavor had a significant effect on the hookah microbiota. Overall, Pseudomonas, Bacillus, Sphingomonas, and Methylobacterium were the predominant bacterial taxa across all products. Additionally, we observed compositional differences between hookah brands across the 14-day incubation. These data suggest that the bacterial communities of hookah tobacco are diverse and differ across brands and flavors, which may have critical implications regarding exposures to specific bacteria among hookah users. Key points • Commercial hookah products harbor diverse bacterial communities. • Brands and flavors impact the diversity of these communities. • Research on their viability and transmission to users’ respiratory tracts is needed. Graphical abstract

A roadmap from unknowns to knowns: Advancing our understanding of the microbiomes of commercially available tobacco products

et al. 2021

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Tobacco smoking is still the leading cause of preventable diseases and death in the USA and throughout the globe. Under Section 904(a)(3) of the US Federal Food, Drug, and Cosmetic Act, tobacco manufacturing companies need to report on quantities of harmful and potentially harmful constituents (HPHCs) in all tobacco products. While the extensive HPHC list of 2012 includes 93 chemicals, which are categorized as carcinogenic, respiratory, cardiovascular, or reproductive toxicants or addictive compounds, it fails to include microorganisms (bacteria and fungi) that have been shown to contribute to adverse health outcomes among tobacco users. Nevertheless, over the last 50 years, researchers have studied microorganisms in a variety of tobacco products using both culture-based and culture-independent techniques. In this mini-review, we provide an overview of this body of research, detailing the bacterial and fungal microbiomes residing in commercial tobacco products. Overall, studies have characterized over 89 unique bacterial genera and 19 fungal genera in cigarettes, cigars, cigarillos, hookah, and smokeless tobacco. The most predominant bacterial genera are Bacillus , Pseudomonas , and Staphylococcus . Fungal genera identified have included Aspergillus , Penicillium , Mucor , Alternaria , Cladosporium , Streptomyces , and Candida , to name a few . While some of the identified microorganisms are known human pathogens, others are potential opportunistic pathogens. Given the vast array of microorganisms that are present across diverse types of tobacco products, future research should be focused on the viability of these microorganisms, as well as their ability to transfer to the user’s respiratory tract, potentially contributing to adverse health outcomes. Key points • Commercial tobacco products harbor diverse bacterial and fungal communities. • Some of these microorganisms are known or opportunistic human pathogens. • Research on their viability and transmission to users’ respiratory tracts is needed.