Identification of bacterial and fungal components in tobacco and tobacco smoke

The microbiological composition of tobacco products was studied using culture and chemical analysis (of tobacco leaves) or chemical analysis only (tobacco and tobacco smoke). The chemical analyses utilized gas chromatography-tandem mass spectrometry for determining 3-hydroxy fatty acids, muramic acid, and ergosterol as markers of respectively lipopolysaccharide (LPS), peptidoglycan, and fungal biomass. Mesophilic bacteria dominated in both fresh and cured tobacco leaves; a range of additional bacteria and fungi were also found albeit in minor amounts. The peptidoglycan and LPS concentrations were approximately the same in tobacco leaves as in cigarette tobacco. The concentrations of the measured microbial components were much lower in some cigarettes locally produced in China, Korea, and Vietnam than in cigarettes of international brands purchased in the same countries, and the concentrations in the smoke were in general agreement with the concentrations in cigarette tobacco. No differences in microbial load in tobacco of "light" and "full flavor" cigarettes were seen. Storing cigarettes at high humidity resulted in elevated levels of fungi in the cigarette tobacco leading to increased ergosterol concentrations in the smoke. The fact that tobacco smoke is a bioaerosol may help to explain the high prevalence of respiratory disorders among smokers and non-smokers exposed to second hand smoke since the same symptoms are also commonly associated with exposure to bioaerosols

Age influence on mice lung tissue response to <i>Aspergillus fumigatus</i> chronic exposure

[b]Introduction and objective[/b]. Exposure to conidia of [i]Aspergillus fumigatus[/i] was described as a causative factor of a number of the respiratory system diseases, including asthma, chronic eosinophilic pneumonia, hypersensitivity pneumonitis and bronchopulmonary aspergillosis. The study investigates the effects of the repeated exposure to [i]A. fumigatus[/i] in mice pulmonary compartment. Our work tackles two, so far insufficiently addressed, important aspects of interaction between affected organism and[i] A. fumigatus[/i]: 1) recurrent character of exposure (characteristic for pathomechanism of the abovementioned disease states) and 2) impact of aging, potentially important for the differentiation response to an antigen. [b]Materials and methods[/b]. In order to dissect alterations of the immune system involved with both aging and chronic exposure to [i]A. fumigatus[/i], we used 3- and 18-month-old C57BL/6J mice exposed to repeated[i] A. fumigatus[/i] inhalations for 7 and 28 days. Changes in lung tissue were monitored by histological and biochemical evaluation. Concentration of pro- and anti-inflammatory cytokines in lung homogenates was assessed by ELISA tests. [b]Results and conclusions. [/b]Our study demonstrated that chronic inflammation in pulmonary compartment, characterized by the significant increase of proinflammatory cytokines (IL1, IL6, IL10) levels, was the dominant feature of mice response to repeated [i]A. fumigatus[/i] inhalations. The pattern of cytokines' profile in the course of exposure was similar in both age groups, however in old mice the growth of the cytokines' levels was more pronounced (especially in case of IL1)

Evaluation of activities aimed at preventing microbiological risks in dental practice

Background: Microbiological contamination of water in dental unit waterlines (DUWL) creates a risk of cross-infections, and is a source of biological risk factors in the work environment of a dentist. The aim of the study was to evaluate dentists' knowledge on DUWL microbiological contamination and the scope of activities/procedures they undertake to monitor it. Material and Methods: The questionnaire survey was conducted in 2010 among 107 Polish dentists using dental units in everyday clinical practice. Results: It has been found that in their daily practice, dentists do not follow procedures leading to reduction or elimination of microbiological contamination of dental unit reservoir water. They are not aware of microbiological contamination of DUWL that supply working handpieces with water. They are unaware of the principles of dealing with dental water and water supply systems or the health risk posed by microbiological contamination of unit water for a dental team and patients. Conclusions: It is necessary to provide dentists with information on microbiological contamination of water in dental units, on the correct procedures of handling water and waterlines that supply working handpieces with water. Med Pr 2013;64(1):11–1

Levels of bacterial endotoxin in air of animal houses determined with the use of gas chromatography - Mass spectrometry and Limulus test

Air samples were collected on glass fibre filters in 22 animal houses and 3 hay storage barns and examined for the presence of bacterial endotoxin with the Limulus (LAL) test and the gas chromatography - tandem mass spectrometry (GC-MSMS) technique, based on detection of 3-hydroxy fatty acids (3-OH-FAs) as chemical markers of the endotoxin lipopolysaccharide. The median concentrations of airborne endotoxin determined with LAL test in poultry houses, sheep sheds, piggeries, cow barns, and horse stables were respectively 62.49 mu g/m(3), 26.2 mu g/m(3), 3.8 mu g/m(3), 1.65 mu g/m(3), and 1. 14 mu g/m(3), while those determined with the GC-MSMS technique were respectively 1.06 mu g/m(3), 7.91 mu g/m(3), 0.2 mu g/m(3), 0.31 mu g/m(3), and 1.42 mu g/m(3). The median concentrations of airborne endotoxin determined with LAL test and GC-MSMS technique in hay storage barns were much smaller, 0.09 mu g/m(3) and 0.03 mu g/m(3), respectively. The concentrations of airborne endotoxin (LPS) detected with GC-MSMS method in the air of sheep sheds were significantly greater than in all other examined facilities, while those detected in hay storage barns were significantly smaller than in all other examined facilities (p<0.05). The concentrations of airborne endotoxin determined with LAL test and GC-MSMS analysis exceeded in most of animal houses examined (91% by each method) the threshold limit value for airborne endotoxin of 0.1 mu g/m(3) proposed by various authors. A significant correlation (p<0.05) between the concentrations of endotoxin determined with the LAL and GC-MSMS techniques was found in the air samples collected in poultry houses and sheep sheds, but not in other examined facilities. 3-OH FAs with C-14-C-18 chains were predominant in the air of the facilities under study. A significant correlation (p<0.05) was found between the concentrations of endotoxin determined with LAL test and the amounts of 3-OH FAs with C-14-C-16 chains. In conclusion, endotoxin in the concentrations detected in this study may present a respiratory hazard to both humans and livestock animals

Levels Of Bacterial Endotoxin In The Samples Of Settled Dust Collected In Animal Houses

Samples of settled dust were collected in 14 animal houses and examined for the presence of bacterial endotoxin with the Limulus (LAL) test and the gas chromatography - tandem mass spectrometry (GC-MSMS) technique, based oil the detection of 3-hydroxy fatty acids (3-OH-FAs) as chemical markers of the endotoxin lipopolysaccharide. The median concentrations of the endotoxin in dust determined with LAL test in sheep sheds, poultry houses, and horse stables were 15,687.5 mu g/g, 8,081.8 mu g/g, and 79.3 mu g/g, respectively, while those determined with the GC-MSMS technique were 868.0 mu g/g, 580.0 mu g/g, and 496.0 mu g/g, respectively. Statistical comparison of the results yielded with LAL test and GC-MSMS technique revealed a weak correlation between both methods. Fatty acids with 14-16 carbon chains (3-OH-C-14 and 3-OH-C-16) were predominant in the settled dust of the facilities under study. In conclusion, endotoxin in the concentrations detected in this study may present it respiratory hazard to both livestock animals and farm workers. Thus, the prevention measures aiming to lower the exposure to endotoxin in livestock facilities are highly desirable

Seasonal microbiological quality of air in veterinary practices in Poland

Numerous studies focused on the bioaerosols in the areas of industry, agriculture and animal husbandry, concerning both residential and public buildings, have been conducted continuously for many years. The aim of the present work was to determine the concentration and composition of mesophilic bacterial flora in the air of selected medical and veterinary clinics located in the cities and in the countryside. Air sampling was carried out in 2011–2013 in 44 veterinary practices in autumn-winter and spring-summer seasons. The concentration of bacteria ranged from 39 – 5,034 cfu/m 3 , with higher values recorded in offices operating in the cities. In the examined medical and veterinary offices, Gram-positive bacteria comprised the largest group of microorganisms, among which Gram-positive cocci of the genus Staphylococcus prevailed, with the highest average of 1,074.40 cfu/m 3 in urban offices during the autumn season. The smallest group was represented by Gram-negative bacteria, with a concentration of 0.0 – 215 cfu/m 3 . In total, 93 kinds/species of bacteria were identified. A 12-month series of studies showed the highest mean concentrations of microorganisms in autumn for offices located in the city, while the lowest in winter for rural centres. The environment of veterinary offices is a habitat of pathogenic and potentially pathogenic bacteria, which may pose health problems not only for residents, but also for the animals

Respiratory Disorders In Two Workers Of Customs Depositories Occupationally Exposed To Mouldy Tobacco

Work-related respiratory symptoms, including dyspnoea, cough, fever, tiredness and malaise, were recorded in two customs officers employed in 2 depositories of confiscated cigarettes, of which one showed signs of dampness. Microbiological sampling of the air and the cigarettes stored in a damp depository revealed the presence of potentially pathogenic fungi and bacteria and the biochemical markers of bacterial lipopolysaccharide and fungal biomass. The Penicillium species (P. simplicissimum, P. inflatum, P commune) dominated in the damp depository, while in the other one Aspergillus fumigatus was prevalent. The patients under study did not show a specific sensitization to microbial allergens in the precipitin test, the test for inhibition of leukocyte migration and the bronchial provocation challenge, except for a weak reaction to fungal allergens in the test for inhibition of leukocyte migration. Moreover, one patient responded with subjective symptoms after exposure to inhalation of increased doses of Penicillium simplicissimum antigen. Both cases were diagnosed as a specific form of organic dust toxic syndrome (ODTS). It is hypothesized that the symptoms were evoked most probably by the non-specific action of low molecular fungal metabolites, such as mycotoxins or VOCs (volatile organic compounds), with the possible contribution of bacterial endotoxin. However, as there is no a direct proof to support this presumption, and the effects of nicotine and other tobacco constituents cannot be excluded, further studies are needed to elucidate etiopathogenesis of the disorders associated with the exposure to stored tobacco

Occupational exposure to organic dust, microorganisms, endotoxin and peptidoglycan among plants processing workers in Poland

The objective of present work was to determine and compare the components of bioaerosol in several sectors of plant processing industries. The Study was conducted in 10 Facilities engaged in herb and grain processing, flax threshing, grain storing, baking, and cereals production. The air samples were taken on glass fibre litters with an AS-50 sampler. We determined the concentrations of airborne microorganisms, dust, endotoxin and peptidoglycan. Total concentrations of viable airborne microorganisms ranged from 0.18-861.4 x 10(3) cfu/m(3). The highest levels of microbial contamination of the air were observed at flax farms, in grain elevators and in a herb processing plant. Gram-positive bacteria aid fungi were detected at all sampling sites, and their median concentrations were respectively 18.1 x 10(3) cfu/m(3) and 0.66 x 10(3) cfu/m(3). The concentration of Gram-negative bacteria ranged from 0.0-168.0 x 10(3) cfu/m(3). The concentration of thermophilic actinomycetes ranged from 0.0-1.45 x 10(3) cfu/m(3). Qualitatively Gram-positive bacteria constituted 23-93% of the total microbial Count. The most common species were: Staphylococcus spp., Curtobacterium pusillum, Rhodococcus fascians, Aureobacterium testaceum, Sanguibacter keddieii, Microbacterium spp., and Bocillus spp. Gram-negative bacteria formed 0-48% of the total Count. The species Pantoea agglomerans dominated in all examined air samples. Fungi constituted 2.5-76.9% of the total microbial count. Among them, Penicillium spp., Mucor spp., Alternario spp., Aspergillus niger, and Aspergillus spp. were found. The dust concentration ranged from 0.18-86.9 mg/m(3). The concentration of endotoxin was large and ranged front 0.0041-1562.6 mu g/m(3). Muramic acid, the chemical marker of peptidoglycan, was detected in 9 out of 13 (69.2%) collected samples. The concentration of peptidoglycan ranged front 1.93-416 ng/m(3). A highly significant correlation was found between the individual components of bioaerosol determined in this study. The concentration of endotoxin was correlated with the concentration of Gram-negative bacteria, total microorganisms, and peptidoglycan (R>0.9, p0.9, p<0.001)

Mouse model of hypersensitivity pneumonitis after inhalation exposure to different microbial antigens associated with organic dusts.

The aim of this study was to reproduce in laboratory conditions hypersensitivity pneumonitis (HP) pathogenesis in a new animal model predictive of the human response, and to select the microbial antigen associated with organic dust that exerts the strongest pathogenic effect on the respiratory organ. To achieve this goal, mice of the strain C57BL/6J prone to fibrosis were exposed for 1 hour daily up to 28 days to the inhalation of aerosols of each of the 5 microbial components of organic dusts whose conjunction with the occurrence of HP has been confirmed by numerous authors: Pantoea agglomerans saline extract (SE), P. agglomerans microvesicle-bound endotoxin, Saccharopolyspora rectivirgula SE, Aspergillus fumigatus SE, saline extract of dust from a grain sample overgrown with S. rectivirgula and Thermoactinomyces vulgaris, and a saline solvent (PBS) was used as a control. Exposure of the animals to organic dust components was conducted using a novel inhalation challenge set. Lung samples were collected from untreated mice and from mice exposed for 7 and 28 days, and examined by digitalized histopathology and biochemistry for the presence of inflammatory changes and fibrosis. P. agglomerans SE appeared to be the sole antigen which evoked a statistically significant fibrosis and a significant increase of hydroxyproline in the lungs of mice exposed for 28 days to this extract, both compared to the mice untreated and to those exposed to the solvent. P. agglomerans SE also evoked the strongest and statistically significant inflammatory response in the lungs of the mice, both after 7 and 28 days of exposure. After 7 days, significant inflammatory changes were also found in mice exposed to A. fumigatus SE, and after 28 days in mice exposed to all antigens. In conclusion, our results allow us to define a useful animal model of HP which can be a supplement for now commonly used bleomycin model. This model should comprise: present set of instruments for inhalation, mice of the line C57BL/6J and the saline extract of P. agglomerans as the antigen. For a better understanding of the presented results, a detailed study covering immunological investigations, focused on the mechanism of antigen action, are needed

Mouse model of hypersensitivity pneumonitis after inhalation exposure to different microbial antigens associated with organic dusts.

The aim of this study was to reproduce in laboratory conditions hypersensitivity pneumonitis (HP) pathogenesis in a new animal model predictive of the human response, and to select the microbial antigen associated with organic dust that exerts the strongest pathogenic effect on the respiratory organ. To achieve this goal, mice of the strain C57BL/6J prone to fibrosis were exposed for 1 hour daily up to 28 days to the inhalation of aerosols of each of the 5 microbial components of organic dusts whose conjunction with the occurrence of HP has been confirmed by numerous authors: Pantoea agglomerans saline extract (SE), P. agglomerans microvesicle-bound endotoxin, Saccharopolyspora rectivirgula SE, Aspergillus fumigatus SE, saline extract of dust from a grain sample overgrown with S. rectivirgula and Thermoactinomyces vulgaris, and a saline solvent (PBS) was used as a control. Exposure of the animals to organic dust components was conducted using a novel inhalation challenge set. Lung samples were collected from untreated mice and from mice exposed for 7 and 28 days, and examined by digitalized histopathology and biochemistry for the presence of inflammatory changes and fibrosis. P. agglomerans SE appeared to be the sole antigen which evoked a statistically significant fibrosis and a significant increase of hydroxyproline in the lungs of mice exposed for 28 days to this extract, both compared to the mice untreated and to those exposed to the solvent. P. agglomerans SE also evoked the strongest and statistically significant inflammatory response in the lungs of the mice, both after 7 and 28 days of exposure. After 7 days, significant inflammatory changes were also found in mice exposed to A. fumigatus SE, and after 28 days in mice exposed to all antigens. In conclusion, our results allow us to define a useful animal model of HP which can be a supplement for now commonly used bleomycin model. This model should comprise: present set of instruments for inhalation, mice of the line C57BL/6J and the saline extract of P. agglomerans as the antigen. For a better understanding of the presented results, a detailed study covering immunological investigations, focused on the mechanism of antigen action, are needed