Chopyk Jessica, Chattopadhyay Suhana, Kulkarni Prachi, Smyth Eoghan M, Hittle Lauren E, Paulson Joseph N, Pop Mihai, Buehler Stephanie S, Clark Pamela I, Mongodin Emmanuel F, Sapkota Amy R
Maryland Institute for Applied Environmental Health, School of Public Health, University of Maryland College Park, MD, USA.
Maryland Institute for Applied Environmental Health, School of Public Health, University of MarylandCollege Park, MD, USA; Institute for Genome Sciences and Department of Microbiology and Immunology, School of Medicine, University of MarylandBaltimore, MD, USA.
Front Microbiol. 2017 Mar 7;8:358. doi: 10.3389/fmicb.2017.00358. eCollection 2017.
Tobacco products, specifically cigarettes, are home to microbial ecosystems that may play an important role in the generation of carcinogenic tobacco-specific nitrosamines (TSNAs), as well as the onset of multiple adverse human health effects associated with the use of these products. Therefore, we conducted time-series experiments with five commercially available brands of cigarettes that were either commercially mentholated, custom-mentholated, user-mentholated, or non-mentholated. To mimic user storage conditions, the cigarettes were incubated for 14 days under three different temperatures and relative humidities (i.e., pocket, refrigerator, and room). Overall, 360 samples were collected over the course of 2 weeks and total DNA was extracted, PCR amplified for the V3V4 hypervariable region of the 16S rRNA gene and sequenced using Illumina MiSeq. A subset of samples ( = 32) was also analyzed via liquid chromatography with tandem mass spectrometry for two TSNAs: '-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). Comparative analyses of the five tobacco brands revealed bacterial communities dominated by , , and , with relatively stable in abundance regardless of storage condition. In addition, core bacterial operational taxonomic units (OTUs) were identified in all samples and included sp., sp., unknown , sp., sp., , and . Additional OTUs were identified that significantly changed in relative abundance between day 0 and day 14, influenced by brand and storage condition. In addition, small but statistically significant increases in NNN levels were observed in user- and commercially mentholated brands between day 0 and day 14 at pocket conditions. These data suggest that manufacturing and user manipulations, such as mentholation and storage conditions, may directly impact the microbiome of cigarette tobacco as well as the levels of carcinogens.
烟草制品，尤其是香烟，是微生物生态系统的宿主，这些微生物生态系统可能在致癌性烟草特有亚硝胺（TSNAs）的产生以及与使用这些产品相关的多种不良人类健康影响的发生中发挥重要作用。因此，我们对五个市售品牌的香烟进行了时间序列实验，这些香烟分别是商业薄荷醇化的、定制薄荷醇化的、用户薄荷醇化的或非薄荷醇化的。为了模拟用户储存条件，将香烟在三种不同温度和相对湿度（即口袋、冰箱和房间）下孵育14天。总体而言，在2周的时间内收集了360个样本，提取了总DNA，对16S rRNA基因的V3V4高变区进行PCR扩增，并使用Illumina MiSeq进行测序。还通过液相色谱-串联质谱法对一部分样本（n = 32）进行了两种TSNAs的分析：N'-亚硝基降烟碱（NNN）和4-（甲基亚硝胺基）-1-（3-吡啶基）-1-丁酮（NNK）。对这五个烟草品牌的比较分析表明，细菌群落以芽孢杆菌属、葡萄球菌属和假单胞菌属为主，无论储存条件如何，芽孢杆菌属的丰度相对稳定。此外，在所有样本中都鉴定出了核心细菌操作分类单元（OTUs），包括芽孢杆菌属、葡萄球菌属、未知菌属、假单胞菌属、寡养单胞菌属、不动杆菌属和嗜麦芽窄食单胞菌属。还鉴定出了其他OTUs，其相对丰度在第0天和第14天之间因品牌和储存条件而有显著变化。此外，在口袋条件下，在第0天和第14天之间，用户薄荷醇化品牌和商业薄荷醇化品牌中的NNN水平有小幅但具有统计学意义的升高。这些数据表明，制造和用户操作，如薄荷醇化和储存条件，可能直接影响卷烟烟草的微生物群以及致癌物水平。
