Middle age enhances expression of innate immunity genes in a female mouse model of pulmonary fibrosis.

The lungs are highly sensitive to tissue fibrosis, with a clear age-related component. Among the possible triggers of pulmonary fibrosis are repeated inhalations of fine organic particles. How age affects this response, is still far from being fully understood. We examined the impact of middle-age on gene expression in pulmonary fibrosis, using the novel "inhalation challenge set" mouse model. Our results demonstrate that the response of female mice to exposure of Pantoea agglomerans extract primarily involves various immune-related pathways and cell-cell/cell-extracellular matrix interactions. We found that middle-age had a strong effect on the response to the P. agglomerans-induced lung fibrosis, featured by a more rapid response and increased magnitude of expression changes. Genes belonging to innate immunity pathways (such as the TLR signaling and the NK-cell mediated cytotoxicity) were particularly up-regulated in middle-aged animals, suggesting that they may be potential targets for the treatment of pulmonary fibrosis caused by inhalations of organic particles. Our analysis also highlights the relevance of the "inhalation challenge set" mouse model to lung aging and related pathology.

Pantoea agglomerans: a mysterious bacterium of evil and good. Part II--Deleterious effects: Dust-borne endotoxins and allergens--focus on grain dust, other agricultural dusts and wood dust.

Pantoea agglomerans, a Gram-negative bacterium developing in a variety of plants as epiphyte or endophyte is particularly common in grain and grain dust, and has been identified by an interdisciplinary group from Lublin, eastern Poland, as a causative agent of work-related diseases associated with exposure to grain dust and other agricultural dusts. The concentration of P. agglomerans in grain as well as in the settled grain and flour dust was found to be high, ranging from 10(4)-10(8) CFU/g, while in the air polluted with grain or flour dust it ranged from 10(3)-10(5) CFU/m(3) and formed 73.2-96% of the total airborne Gram-negative bacteria. The concentration of P. agglomerans was also relatively high in the air of the facilities processing herbs and other plant materials, while it was lower in animal farms and in wood processing facilities. Pantoea agglomerans produces a biologically-potent endotoxin (cell wall lipopolysaccharide, LPS). The significant part of this endotoxin occurs in dusts in the form of virus-sized globular nanoparticles measuring 10-50 nm that could be described as the 'endotoxin super-macromolecules'. A highly significant relationship was found (R=0.804, P=0.000927) between the concentration of the viable P. agglomerans in the air of various agricultural and wood industry settings and the concentration of bacterial endotoxin in the air, as assessed by the Limulus test. Although this result may be interfered by the presence of endotoxin produced by other Gram-negative species, it unequivocally suggests the primary role of the P. agglomerans endotoxin as an adverse agent in the agricultural working environment, causing toxic pneumonitis (ODTS). Numerous experiments by the inhalation exposure of animals to various extracts of P. agglomerans strains isolated from grain dust, including endotoxin isolated with trichloroacetic acid (LPS-TCA), endotoxin nanoparticles isolated in sucrose gradient (VECN), and mixture of proteins and endotoxin obtained by extraction of bacterial mass in saline (CA-S), showed the ability of these extracts to evoke inflammatory and fibrotic changes in the lungs, to stimulate alveolar macrophages to produce superoxide anion (O2(-)), interleukin-1 (IL-1) and chemotactic factors for other macrophages and neutrophils, and to increase the pulmonary concentrations of toll-like receptors and chemokines. The most potent properties showed the CA-S which may be attributed to the allergenic properties of P. agglomerans proteins enhanced by the presence of the autologous endotoxin. The results of these experiments are in accord with the clinical studies which revealed a high reactivity of the agricultural and grain industry workers to allergenic extracts of P. agglomerans, and the presence in these populations of hypersensitivity pneumonitis and asthma cases caused by this bacterium. P. agglomerans has been also identified as a potential causative agent of allergic dermatitis in farmers and of allergic pulmonary disorders in cattle. In conclusion, similar to the cotton industry, also in the grain industry and in agriculture, Pantoea agglomerans should be regarded as one of the major causative agents of work-related diseases, caused by the adverse effects of protein allergens and endotoxin produced by this bacterium.

Relationship between concentrations of microbiological agents in the air of agricultural settings and occurrence of work-related symptoms in exposed persons.

For assessment of the dose-response relationship between concentrations of microbial agents in the air of various agricultural settings and occurrence of work-related symptoms in exposed workers, a meta-analysis of the results obtained in 1994-2007 on the territory of eastern Poland was performed. The studies on the airborne concentrations of total culturable microorganisms, mesophilic bacteria, Gram-negative bacteria, thermophilic actinomycetes, fungi, and bacterial endotoxins, as well as on the frequency of work-related respiratory and general symptoms in the exposed workers, were carried out at grain, thyme, valerian, flax, and hop handling on farms, in cow barns, piggeries, horse stables and in a modern hatchery. The airborne concentrations of the total microorganisms were in the range of 9.2-1236.5 × 10(3) CFU/m(3) , of the total mesophilic bacteria 3.5-1225.8 × 10(3) CFU/m(3) , of Gram-negative bacteria 0.0- 46.2 × 10(3) CFU/m(3) , of thermophilic actinomycetes 0.0-7.1 × 10(3) CFU/m(3) , of fungi 2.1-77.9 × 10(3) CFU/m(3) , and of bacterial endotoxin 0.00925-429.55 µg/m(3) . The frequency of work-related symptoms ranged between 21.7-63.8%. In a meta-analysis for assessment of the correlations between the log-transformed concentrations of airborne microbial agents and the occurrence of work-related symptoms, the multiple regression test was applied. Statistically significant correlations were found between the occurrence of work-related symptoms and the concentration of total airborne microorganisms (R=0.748555; P=0.020317), mesophilic bacteria (R=0.7573; P=0.029548), Gram-negative bacteria (R=0.835938; P=0.019129), and endotoxins (R=0.705356; P=0.03378). The correlations between the concentrations of thermophilic actinomycetes and fungi, on one side, and frequency of work-related symptoms on the other, did not attain the threshold of significance (P=0.087049 and P=0.062963, respectively). Results of the meta-analysis confirm harmful health effects of the total airborne microorganisms, total airborne mesophilic bacteria, airborne Gram-negative bacteria, and airborne bacterial endotoxin on the occupationally-exposed agricultural workers, and indicate a need for the establishment of internationally recognized occupational exposure limits for these microbial agents.

Age influence on mice lung tissue response to Aspergillus fumigatus chronic exposure.

INTRODUCTION AND OBJECTIVE: Exposure to conidia of Aspergillus fumigatus 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 A. fumigatus in mice pulmonary compartment. Our work tackles two, so far insufficiently addressed, important aspects of interaction between affected organism and A. fumigatus: 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. MATERIALS AND METHODS: In order to dissect alterations of the immune system involved with both aging and chronic exposure to A. fumigatus, we used 3- and 18-month-old C57BL/6J mice exposed to repeated A. fumigatus 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. RESULTS AND CONCLUSIONS: 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 A. fumigatus 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).

Age influence on hypersensitivity pneumonitis induced in mice by exposure to Pantoea agglomerans.

Hypersensitivity pneumonitis (HP) represents the immunologically mediated lung disease induced by repeated inhalations of a wide variety of certain finely dispersed organic antigens. In susceptible subjects, these inhalations provoke a hypersensitivity reaction characterized by intense inflammation of the terminal bronchioles, the interstitium and the alveolar tree. The inflammation often organizes into granulomas and may progress to pulmonary fibrosis. Our previous work indicated that cell extract of gram-negative bacteria Pantoea agglomerans (SE-PA) causes, in young C57BL/6J mice, pulmonary changes that are very similar to the clinical manifestations of HP in men. The purpose of presented studies was to describe the response of mice immune system while exposed to SE-PA. Particular attention was paid to examine the age influence on SE-PA induced inflammation and fibrosis in lung tissue. We used 3- and 18-month-old C57BL/6J mice. Lung samples were collected from untreated mice and animals exposed to harmful agent for 7 and 28 days. HP development was monitored by histological and biochemical evaluation. Using ELISA tests, we examined concentration of pro- and anti-inflammatory cytokines in lung homogenates. Our study demonstrated again that SE-PA provokes in mice changes typical for the clinical picture of HP, and that successive stages of disease (acute, subacute and chronic) might be obtained by modulation of time exposure. Furthermore, we found that animals' age at the time of sensitization influences the nature of observed changes (cytokine expression pattern) and the final outcome (reaction intensity and scale of fibrosis).

Exposure to bioaerosols in the selected agricultural facilities of the Ukraine and Poland - a review.

The aim of this work was to review the studies on bioaerosols which were carried out in the years 1972-2009 in following branches of agricultural industry in the Ukraine and Poland: animal farms for cows and pigs, animal feed facilities, production of biofuel from rape, herb farms and herb processing facilities. In all facilities were determined: concentration of dust and microorganisms in the air and species composition of microflora. Moreover, in Polish animal farms, herb farms and herb processing facilities, as well as in the Ukrainian animal feed facilities, was determined the concentration of bacterial endotoxin in the air. Dust concentrations in animal farms located in the Ukraine and Poland ranged from 6-200 mg/m(3) and from 0.25-14.05 mg/m(3), respectively, while in animal feed facilities they ranged from 35-306 mg/m(3) and from 3.8-405 mg/m(3), respectively. Dust concentrations in the facilities producing biofuel from rape in the Ukraine were in the range 3.6-28 mg/m(3), whereas on herb farms and in herb processing facilities in Poland they were in the range 0.8- 1,319.6 mg/m(3), and 2.2-946 mg/m(3), respectively. The determined values exceeded in most cases the maximal acceptable concentration (MAC) which in the Ukraine and Poland is equal to 4 mg/m(3). The concentrations of microorganisms on animal farms located in the Ukraine and Poland ranged from 5.5 x 10(4)- 1.9 x 10(7) cfu/m(3) and from 4.7 x 10(4)-1.5 x 10(6) cfu/m(3), respectively, while in animal feed facilities they ranged from 2.7 x 10(4)-2.6 x 10(9) cfu/m(3) and from 1.7 x 10(3)-2.0 x 10(6) cfu/m(3), respectively. Concentrations of microorganisms in the facilities producing biofuel from rape in the Ukraine were in the range 1.5 x 10(3)-5.7 x 10(6) cfu/m(3), whereas on herb farms and in herb processing facilities in Poland they were in the ranges of 8.8 x 10(2)-8.0 x 10(6) cfu/m(3), and 9.7 x 10(3)-6.3 x 10(5) cfu/m(3), respectively. The determined values exceeded in most cases the maximal acceptable concentration (MAC) which in the Ukraine is equal to 5.0 x 10(4) cfu/m(3). The concentrations of endotoxin in Polish animal houses were in the range 0.00125-75.0 ug/m(3), whereas on herb farms and herb processing facilities they were in the ranges of 0.0045- 2,448.8 ug/m(3) and 0.2-681.0 ug/m(3), respectively, and in the Ukrainian animal feed facilities were within the range 0.008-240.0 ug/m(3). They exceeded in most cases the level of 0.2 ug/m(3) proposed as a threshold. In the air microflora of examined facilities prevailed Gram-positive bacteria (corynebacteria, cocci, spore-forming bacilli, actinomycetes) of which some (Arthrobacter spp., thermophilic actinomycetes) could be a cause of allergic alveolitis (hypersensitivity pneumonitis). Among Gram-negative bacteria isolated from the air of agricultural settings dominated the epiphytic species Pantoea agglomerans, possessing potent allergenic and endotoxic properties. Fungi were abundant in the air of the Ukrainian agricultural settings and comprised species able to produce harmful mycotoxins. In conclusion, the airborne biological factors in stated concentrations may exert harmful effects on the state of the health of exposed workers. Formation of the bioaerosol depends on the specificity of the setting, kind of technological operations, degree of mechanization, properties of processed materials, temperature and humidity, and concentration of dust in the air.

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.

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 filters 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 and 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 Bacillus 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., Alternaria 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 from 0.0041-1562.6 microg/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 from 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, p<0.001). The concentration of peptidoglycan was correlated with the concentration of Gram-positive bacteria, Gram-negative bacteria, and total microorganisms (R>0.9, p<0.001).

A novel inhalation challenge set to study animal model of allergic alveolitis.

A novel inhalation challenge set for the study of experimental allergic alveolitis (hypersensitivity pneumonitis) in mice was designed. A finely dispersed aerosol of allergenic extract generated by the commercial ultrasonic nebulizer "TAJFUN MU1" (produced by Medbryt, Warsaw, Poland) was transported to the airtight inhalation chamber. In the chamber were placed 15 perforated containers made of transparent plastic, each containing one mouse. They were coupled in 3 units, each consisted of 5 containers. The constant flow of aerosol through the chamber was assured by commercial vacuum pump "PL 2/3" (AGA LABOR S.C., Warsaw, Poland). The applied set enabled the natural exposure of mice via the inhalation route to known quantities of allergen (usually microbial) suspended in saline, and then dispersed in form of fine aerosol by ultrasonic nebulizer. This method assures the penetration of allergen into the deep parts of lungs, alveoli and bronchioli. The detailed study of histopathological and biochemical changes in the lungs of exposed animals will be the subject of further publications. So far, the retention of endotoxin in the lungs of mice exposed to the extract of a Gram-negative bacterium Pantoea agglomerans and appearance of positive serologic reactions to this extract indicate the effectiveness of the method.

Cathelicidin LL-37, granzymes, TGF-beta1 and cytokines levels in induced sputum from farmers with and without COPD.

The cathelicidin LL-37 is an antimicrobial and lipopolysaccharide neutralizing peptide, possessing pro-inflammatory, tissue repair and remodeling activities. Recent reports indicate that the progression of COPD might be connected with increased levels of LL-37. The numerous experimental data show the potential role of LL-37 in the response to the exposure to organic dust (containing lipopolysaccharide and microorganisms) which is one of the major COPD causative factors. This work strives to further prove the role of LL-37 in the development of COPD. A cross-sectional study was conducted in 30 farmers in the early stages of COPD according to GOLD, 36 healthy farmers and 16 healthy urban dwellers. Collection of induced-sputum samples and lung function testing were conducted before and after work. The quantification of the LL-37 in sputum samples was performed by mass spectrometry and radioisotope techniques. Levels of granzymes A and B, IL-8, IFN-gamma and TGF-beta1 in sputum were measured by ELISA technique. Statistical analysis was conducted by Kruskal-Wallis and Mann-Whitney U tests. Significantly higher levels of LL-37 were observed in sputum samples from farmers with COPD compared to healthy individuals. The concentration of LL-37 in sputum from farmers was significantly higher compared to urban dwellers. The same was true for both granzymes A and B. The results of this study suggest that LL-37 and granzymes A and B may add to the development of COPD. The results suggest also their role in an organism's response to organic dust exposure.

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.

Study on Lyme borreliosis focus in the Lublin region (eastern Poland).

A suburban focus of Lyme borreliosis situated 11 km from the southern border of the city of Lublin (eastern Poland) was characterized. The focus covers an area of circa 100 km(2), surrounding 3 localities inhabited by circa 7,500 people engaged mostly in farming. It was demonstrated that on the area of focus the infection rate of Ixodes ricinus ticks with Borrelia burgdorferi, frequency of serological response of inhabitants to the antigen of Borrelia burgdorferi, and incidence of Lyme borreliosis were significantly (p<0.001) greater compared to the whole territory of Lublin province, and were respectively 13.1 % vs. 4.7 %, 33.0 % vs. 13.7 %, and 0.002 % vs. 0.00075 % .

Efficacy of a novel biofilter in hatchery sanitation: I. Removal of airborne bacteria, dust and endotoxin.

A novel biofilter containing organic, bentonite and halloysite media was applied for elimination of microbial pollutants from the air of an industrial hatchery. The concentrations of total mesophilic bacteria, Gram-negative bacteria, thermophilic actinomycetes, dust and bacterial endotoxin were determined in the air of hatchery during 2 months before installation of the biofilter, and during 6 months after installation of the biofilter, at the inlet and outlet ducts from each medium. Before installation of the biofilter, the concentrations of total mesophilic bacteria, Gram-negative bacteria, thermophilic actinomycetes, dust and endotoxin in the air were within the ranges of 0.97-131.2x10(3) cfu/m3, 0.0-34.4x10(3) cfu/m3, 0.0-0.02x10(3) cfu/m3, 0.37-4.53 mg/m3, and 50.9-520,450.4 ng/m3, respectively. Enterococcus faecalis and Gram-negative bacteria (Acinetobacter spp., Escherichia coli, Enterobacter cloacae, and other species) prevailed among bacterial species recovered from the air of the hatchery. A total of 56 species or genera of bacteria were identified in the air samples taken in the examined hatchery; of these, 11, 11 and 6 species or genera respectively were reported as having allergenic, immunotoxic and/or infectious properties The concentrations of total mesophilic bacteria, Gram-negative bacteria, Enterococcus faecalis and endotoxin found at the inlet duct of the biofilter after its installation were significantly smaller compared to those recorded before its installation (p<0.05). The concentrations of Gram-negative bacteria, Enterococcus faecalis and dust found at the outlet ducts of biofilter after its installation were significantly smaller compared to those recorded at the inlet duct of the biofilter (p<0.01). The concentrations of total meso-philic bacteria were also smaller at the outlet ducts of the biofilter compared to that at the inlet duct; however, the difference was not significant because of the massive growth of Streptomyces species in the biofilter's media which contaminated the outcoming air. In conclusion, the applied biofilter proved to be effective in the elimination of potentially pathogenic bacteria, dust and endotoxin from the air of the hatchery. The efficacy of the biofilter could be improved by the inhibition of the Streptomyces growth in the media of the biofilter.

Efficacy of a novel biofilter in hatchery sanitation: II. Removal of odorogenous pollutants.

The present research assessed the treatment efficiency of odorogenous pollutants in air from a hatchery hall vented on organic and organic-mineral beds of an enclosed-container biofilter. In this study, the following media were used: organic medium containing compost and peat (OM); organic-mineral medium containing bentonite, compost and peat (BM); organic-mineral medium containing halloysite, compost and peat (HM). The concentration of odorogenous gaseous pollutants (sulfur compounds and amines) in the hatching room air and in the air after biotreatment were determined by gas chromatography. In the hatchery hall among the typical odorogenous pollutants, there were determined 2 amines: 2-butanamine and 2-pentanamine, hydrogen sulfide, sulfur dioxide, carbon disulfide, sulfides and mercaptans. Ethyl mercaptan showed the highest levels as its mean concentration in the hatchery hall air exceeded 60 microg/m3 and in single samples even 800 microg/m3. A mean concentration of 2-butanamine and sulfur dioxide in the examined air also appeared to be relatively high--21.405 microg/m3 and 15.279 microg/m3, respectively. In each filter material, the air treatment process ran in a different mode. As the comparison reveals, the mean reduction of odorogenous contaminants recorded in the hall and subjected to biotreatment was satisfying as it surpassed 60% for most established pollutants. These high removal values were confirmed statistically only for single compounds. However, a low removal level was reported for hydrogen sulfide and sulfur dioxide. No reduction was recorded in the bentonite supplemented medium (BM) for sulfur dioxide and methyl mercaptan. In the organic medium (OM) no concentration fall was noted for dipropyl sulfide either. In all the media investigated, the highest removal rate (100%), not confirmed statistically, was observed for carbon disulfide. Very good results were obtained in the medium with a bentonite additive (BM) for both identified amines, whose mean elimination rate exceeded 60% (p

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 microg/m3, 26.2 microg/m3, 3.8 microg/m3, 1.65 microg/m3, and 1.14 microg/m3, while those determined with the GC-MSMS technique were respectively 1.06 microg/m3, 7.91 microg/m3, 0.2 microg/m3, 0.31 microg/m3, and 1.42 microg/m3. The median concentrations of airborne endotoxin determined with LAL test and GC-MSMS technique in hay storage barns were much smaller, 0.09 microg/m3 and 0.03 microg/m3, 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 microg/m3 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 C14-C18 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 C14-C16 chains. In conclusion, endotoxin in the concentrations detected in this study may present a respiratory hazard to both humans and livestock animals.

Health effects of exposure to organic dust in workers of a modern hatchery.

The aim of the presented study was to determine the health status of workers occupationally exposed to moderate amounts of organic dust, employed in a modern hatchery with an efficient ventilation system. A group of 32 hatchery workers was examined. As a reference group, 50 urban dwellers not exposed to any kind of organic dust were examined. All people were interviewed for the presence of work-related symptoms and subjected to physical and spirometric examinations. Blood sera were examined for the presence of precipitins against 13 antigens associated with organic dust, and for the presence of total and chicken-specific No significant differences were found between the spirometric values in the group of hatchery workers and the reference group. Positive precipitin reactions were noted mostly with the antigens of Gram-negative bacteria associated with organic dust. The frequencies of positive reactions to antigens of Escherichia coli and Acinetobacter baumannii in hatchery workers were significantly greater compared to the reference group (p<0.05). Precipitin reactions to Gram-positive non-branching bacteria, actinomycetes, fungi and birdactions to antigens of Escherichia coli and Acinetobacter baumannii in hatchery workers were significantly greater compared to the reference group (p<0.05). Precipitin reactions to Gram-positive non-branching bacteria, actinomycetes, fungi and bird protein, were rare or absent. The mean concentration of total IgE in sera of hatchery workers was nearly 3 times greater compared to the reference group, and the difference proved to be statistically significant (p<0.05). No specific IgE antibodies against chicken feathers were detected in the blood of hatchery workers and referents. In conclusion, the examined hatchery workers showed a moderate frequency of work-related symptoms, no decline in lung function and low reactivity to most microbial and bird protein allergens. These results suggest that the effects of exposure to organic dust in workers of modern hatcheries with an efficient ventilation system are less compared to the workers of poultry farms, such as broiler or egg laying houses.

Respiratory effects of exposure to dust from herbs.

A group of 150 people occupationally exposed to dust from herbs were examined. As a reference group, 50 urban dwellers, not exposed to any kind of organic dust were examined. People were subjected to routine physical examination and to lung function examinations with the LUNGTEST 500 spirometer (MES, Kraków, Poland). The spirometric values of the forced expiratory volume in one second (FEV (1)), vital capacity (VC) and FEV(1)/VC were recorded before and after work. Physical chest examination revealed pathological crepitations in 10 people (6.7 %). The mean baseline spirometric values in the study and reference groups did not show significant differences compared to the normal values. In the herb workers exposed to organic dust the post-shift decrease of all analysed spirometric values was noted. The post-shift decrease of some spirometric values (VC, VC % of normal values) was highly significant (p < 0.01). There was evidenced of a significant positive correlation between the age of examined people and decrease of VC and FEV(1) values. In 12 exposed workers the decrease of FEV(1) or FEV(1) % of normal values higher than 15 % was noted. 50 % of these workers cultivated thyme (Thymus vulgaris L.). This may suggest that dust from herbs, especially thyme dust, may cause acute airway obstruction. In the group showing significant decrease of FEV(1)/FEV(1) % of normal values ( > 15 %) the frequency of reported respiratory work-related symptoms (83.3 %) was higher than in the rest of exposed group (61.5 %). In conclusion, occupational exposure to dust from herbs may cause harmful effects on the respiratory system among herb processing workers. This indicates the need for use of prophylactic measures in this professional group, the more so as number of people occupationally exposed to dust from herbs is growing.

Exposure to airborne microorganisms, dust and endotoxin during processing of valerian roots on farms.

The aim of this study was to determine the levels of microorganisms, dust and endotoxin in the air during various stages of valerian (Valeriana officinalis) roots processing by herb farmers and to examine the species composition of airborne microflora. Air samples were collected on glass fibre filters by use of personal samplers on 15 farms owned by valerian cultivating farmers, located in Lublin province (eastern Poland). The concentrations of total viable microorganisms (bacteria + fungi) in the air showed a marked variability and were within a range of 0.95-7,966.6 x 10(3) cfu/m (3). Though median was relatively low (10.75 x 10(3) cfu/m (3)), on 4 farms the concentrations exceeded the level of 10(5) cfu/m (3) and on 1 farm the level of 10(6) cfu/m (3). During the processing of valerian roots, distinct changes could be observed in the composition of airborne microflora. In the first stages of processing, the freshly dug and washed roots until shaking in the drying room, the most numerous were Gram-negative bacteria of the family Pseudomonadaceae (mostly Stenotrophomonas maltophilia, Pseudomonas chlororaphis and Pseudomonas fluorescens). After drying, the dominant organisms were thermo-resistant endospore-forming bacilli (Bacillus spp.) and fungi, among which prevailed Aspergillus fumigatus. Altogether, 29 species or genera of bacteria and 19 species or genera of fungi were identified in the farm air during valerian processing, of these, 10 and 12 species or genera respectively were reported as having allergenic and/or immunotoxic properties. The concentrations of airborne dust and endotoxin on the examined farms were very large and ranged from 10.0-776.7 mg/m (3), and from 0.15-24,448.2 microg/m (3), respectively (medians 198.3 mg/m (3) and 40.48 microg/m (3)). In conclusion, farmers cultivating valerian could be exposed during processing of valerian roots to large concentrations of airborne microorganisms, dust and endotoxin posing a risk of work-related respiratory disease.

Health effects of exposure to herb dust in valerian growing farmers.

The aim of the present study was to determine the health status of farmers cultivating valerian (Valeriana officinalis L.) and occupationally exposed to dust from this plant. A group of 75 valerian growing farmers were examined. As a reference group, 50 urban dwellers, not exposed to any kind of organic dust were examined. All people were interviewed for the presence of work-related symptoms and subjected to physical and spirometric examinations. Skin prick tests were conducted with 4 microbial antigens associated with organic dust and 3 herbal extracts, precipitin tests with 12 microbial antigens and 4 herbal extracts and tests for specific inhibition of leukocyte migration with 4 microbial antigens. 30.7 % of the valerian farmers reported occurrence of work-related symptoms. No significant differences were found between the spirometric values in the group of valerian farmers and the reference group. Valerian farmers showed a low frequency of positive skin reactions to all tested antigens (0-4.0 %), not significantly greater compared to reference group. The frequency of positive precipitin reactions to the antigen of Gram-negative bacterium Pantoea agglomerans was very high in valerian farmers (45.5 %) with 3-fold concentrated sera and significantly greater compared to the reference group (p < 0.001). The positive precipitin response of valerian farmers to other microbial and herbal antigens was much lower or absent and did not show any difference compared to reference group. In the test for specific inhibition of leukocyte migration, the highest frequencies of positive reactions in valerian farmers were noted with Pantoea agglomerans and Saccharopolyspora rectivirgula (15.0 % each), in both cases significantly greater compared to reference group (p < 0.05). In conclusion, the farmers growing valerian showed a moderate frequency of work-related symptoms and low reactivity to most microbial and herbal allergens. They exhibited an increased immunologic response to Gram-negative bacterium Pantoea agglomerans which appears to be the most important risk factor associated with valerian dust.

Exposure of Indian agricultural workers to airborne microorganisms, dust and endotoxin during handling of various plant products.

Microbiological air sampling with the use of personal sampler was performed in 14 small agricultural facilities located in Aurangabad (Southern India), including 5 farms (at harvesting and threshing), 6 godowns (small food storing and processing settings) and 3 grain or cotton mills. In 12 facilities, including harvesting on farms, godowns and mills, concentrations of airborne microorganisms were relatively low, ranging from 1.5 x 10(3) - 8.2 x 10(4) cfu/m(3). The striking rise in the concentrations of airborne microbes, by 2-4 orders of magnitude, was noted on 2 farms during threshing of maize (4.2 x 10(6) cfu/m(3)) and pearl millet (1.1 x 10(7) cfu/m(3)). This was due to release of large quantities of bacteria. During threshing of maize, the most common microorganisms were thermophilic actinomycetes (40 % of the total count) and mesophilic actinomycetes of the genus Streptomyces (39 %), while during threshing of pearl millet the most abundant were corynebacteria (68 %) and Gram-negative bacteria (22 %). Mesophilic bacteria formed 16.7-100 % of the total airborne microflora and were prevalent in 11 out of 14 facilities. Thermophilic actinomycetes and fungi formed respectively 0-43.1 % and 0-83.3 % of the total count and were prevalent in 1 and 2 facilities. The concentrations of airborne dust and endotoxin recorded in the examined agricultural facilities, in most cases were very large and ranged from 2.5-257.5 mg/m(3), and from 0.0625-125.0 microg/m(3), respectively. In 10 out of 14 facilities, the concentrations of dust were of the order 10(1)-10(2) mg/m(3), ranging from 25.0-257.5 mg/m(3). Similarly, in 7 out of 14 facilities the concentrations of endotoxin were of the order 10(1)-10(2) microg/m(3), ranging from 31.25-125.0 microg/m(3). In conclusion, Indian agricultural workers could be exposed during handling of various plant materials to airborne biological hazards posing a risk of work-related respiratory disease. Of these, the most important are: bacterial endotoxin and allergenic species of bacteria and fungi. The greatest risk occurs at threshing of pearl millet and maize which requests an application of the appropriate prevention measures, such as wearing respirators by the workers and using modern threshing machines reducing the dustiness.