Identification of bacterial and fungal components in tobacco and tobacco smoke

Many hundreds of compounds known to contribute to disease development have been identified in tobacco smoke. Both active and second hand smoking causes cancer and a multitude of other diseases such as for example chronic bronchitis and asthma. Three studies [1-3] have revealed that tobacco smoke contains endotoxin (lipopolysaccharide, LPS), a family of inflammatory toxins from Gram-negative bacteria known to cause respiratory disease upon inhalation [4]. Hasday et al. [1] found that the amounts of endotoxin in tobacco were comparable with those of some other agricultural products. While Hasday et al. [1] used a Limulus method for measuring endotoxin, Larsson et al. [2] introduced gas chromatography-tandem mass spectrometry (GC-MSMS) for demonstrating 3-hydroxytetradecanoic acid, a unique LPS constituent [5], in cigarette tobacco and smoke. GC-MSMS is a very specific analysis method for unequivocal identification of LPS. Sebastian et al. [3], using the same GC-MSMS method, demonstrated a linear relationship between the number of cigarettes smoked over a 5-h period indoors and air concentrations of endotoxin. Whether bioactive microbial compounds other than LPS, such as for example peptidoglycan and various fungal components, are present in cigarette smoke is not known from the literature.An integrated method for characterizing microbial composition in environmental samples by GC-MSMS has been developed at our laboratory [6]. The method includes a protocol for preparation and analyzing samples for LPS markers 3-hydroxy fatty acids (3-OH FAs) of 10 – 18 carbon chain lengths, peptidoglycan marker muramic acid (MuAc), and fungal biomass marker ergosterol (Erg). In the present study a modified version [7] of this method was used for 1) measuring 3-OH FAs, MuAc, and Erg in tobacco from cigarettes of international as well as local brands purchased in different countries in Europe and Asia including "light" and "full flavor" cigarettes; 2) analysing the microbiological compo