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PubMed2006

Inflammatory potency of dust from the indoor environment and correlation to content of NAGase and fungi.

The aim of this study was to analyse the contribution of microbial factors to the inflammatory potency of dust (PD). Floor dust was sampled three times from 12 rooms in two schools. The potency of floor dust was measured as interleukin-8 secretion from the lung epithelial cell line A549 after exposure to dust. Measurements of endotoxin, NAGase activity and cultivable fungi in the dust were made. For endotoxin a difference was found between sampling days and for NAGase a difference was found between the schools. The carpeted staff rooms of the two schools had a significantly higher amount of dust/m(2) and endotoxin/m(2), and the PD/m(2) and NAGase activity/m(2) were also higher than in the classrooms with smooth flooring. The PD/m(2) correlated with all fungal parameters except total cultivable fungi. NAGase/m(2) correlated with PD, endotoxin and cultivable Cladosporium sp. per area, and was nearly significantly correlated to total cultivable fungi and Penicillium spp. Therefore; microbiological activity especially from fungi may contribute to the inflammatory potency of floor dust. Carpet flooring may act as a "sink" for microorganisms resulting in a higher inflammatory potency of floor dust, which may reflect building-related symptoms in occupants.

Acetylglucosaminidase↗
PubMed1997

Exposure levels and determinants of inhalable dust exposure in bakeries.

The study's objectives were to measure full-shift exposure to inhalable dust in bakeries and define the determinants of full-shift exposure. Inhalable dust was measured gravimetrically. Ninety-six bakery workers, employed in seven different bakeries, participated in the study. Two side-by-side full-shift inhalable dust samples were obtained from each study participant on a single occasion. Samples were collected on 18 days selected at random. During the entire sampling period, bakers were observed and information on 14 different tasks was recorded at 15 min intervals. Other production characteristics were also recorded for each sampling day. These task and production variables were used in statistical modelling to identify significant predictors of exposure. The mean full-shift inhalable dust exposure was 8.2 mg/m3 (range: 0.1-110 mg/m3). A regression model explained 79% of the variability in exposure. The model indicated that tasks such as weighing, pouring and operating dough-brakers and reversible sheeters increased the exposure, while packing, catching and decorating decreased the exposure. Bread and bun production lines were associated with increased full-shift inhalable dust exposure, while cake production and substitution of dusting with the use of divider oil were associated with decreased exposure. Production tasks and characteristics are strong predictors of personal full-shift exposures to flour dust among bakers; these can be altered to reduce exposure levels.

Analysis of Variance↗
PubMed1990

Depression of alveolar macrophage hydrogen peroxide and superoxide anion release by mineral dusts: correlation with antimony, lead, and arsenic contents.

Activated rabbit alveolar macrophages were incubated with airborne dusts from four West German sites (1 to 200 micrograms/10(6) cells) and waste incinerator fly ash fractions (50 to 500 micrograms/10(6) cells). Quartz dust DQ 12 (5 to 200 micrograms/10(6) cells) and Fe2O3 (0.05 to 50 micrograms/10(6) cells) were used as control dusts. The zymosan-stimulated hydrogen peroxide and superoxide anion release of the macrophages were not affected significantly by Fe2O3. All other investigated dusts decreased the two cell functions which were correlated negatively with surfaces, particle numbers, and antimony, lead, and arsenic contents of the dusts. The influence of heavy metal antagonisms and dust surfaces on dust toxicity against alveolar macrophages is discussed.

Air Pollutants↗
PubMed1986

Effects of the inhalation of dusts from calcium silicate insulation materials in laboratory rats.

The effects of respirable dust from three commercially produced calcium silicate insulation materials were examined in laboratory rats by long-term inhalation and injection techniques. These calcium silicate products have been used as replacements for asbestos in the insulation of the engine rooms of ships, and the particle size distribution of the dust clouds generated for the experimental study closely matched those found in ships during the installation of this type of material. One year of exposure to a dust cloud of 10 mg/m3 of respirable dust had no discernible effect on the length of survival of treated animals compared to controls. No pulmonary lesions were found that appeared associated with the inhalation of calcium silicate per se, but one sample did contain significant amounts of quartz and this did produce a few small pulmonary nodules. While two small pulmonary neoplasms, one malignant and one benign, were found in dusted animals, neither was the cause of death, and the incidence was not significantly different from the control group where no tumors were found. One peritoneal mesothelioma was found in an animal from one of the inhalation groups, but this was considered to be a spontaneous tumor as none of over 100 animals injected intraperitoneally with 25 mg of calcium silicate developed these tumors. While the white blood cell count of dusted animals, compared to controls, was significantly raised in all treated groups at the end of the dusting period, these figures were within the published normal ranges for laboratory rats. It was concluded that the three tested calcium silicate products were harmless to the rats of this species at the doses tested.

Animals↗
PubMed1986

Pulmonary response to impaired lung clearance in rats following excessive TiO2 dust deposition.

Rats were exposed to TiO2 by the inhalation route at concentrations of 0, 10, 50, or 250 mg/m3 for 6 hr/day, 5 days/week for 2 years. Lung weights of rats at 10 mg/m3 were within normal limits after 2 years exposure. Lung weights increased significantly after 6 months at 50 mg/m3 and after 3 months at 250 mg/m3. After 2 years exposure, TiO2 retention in dried lung was 3.1% (26.5 mg per lung) at 10 mg/m3, 16.9% (124 mg per lung) at 50 mg/m3, and 28% (665 mg per lung) at 250 mg/m3. Lung clearance mechanisms appeared to be overloaded at 250 mg/m3. Dust particles were retained in the lung in a dose-related fashion, but there was no significant difference in lung clearance rate between 10 and 50 mg/m3. Lung response at 10 mg/m3 satisfied the biological criteria for a "nuisance dust," while adverse effects resulting from gradually accumulated particles (8.1%, 67.7 mg per lung) were found after 1 year of exposure to 50 mg/m3. An early pulmonary response indicating an overloaded lung clearance mechanism was manifested by massive accumulation of dust-laden macrophages (dust cells), foamy dust cells, free particles, or cellular debris derived from disintegrated foamy dust cells in the alveoli adjacent to the alveolar ducts. Alveolar proteinosis also appeared to be an important marker of an overloaded lung clearance mechanism and was observed at 50 and 250 mg/m3 after 1 year of exposure. Cholesterol granulomas were developed with degenerative foamy dust cells at 50 and 250 mg/m3 after 1 year of exposure. After 2 years exposure at 250 mg/m3, bronchioalveolar adenomas occurred in the alveoli showing type II pneumocyte hyperplasia, while cystic keratinizing squamous carcinomas were developed from squamous metaplasia of alveoli showing bronchiolarization in the alveolar duct region. Since the lung tumors were a unique type of experimentally induced tumors under exaggerated exposure conditions and have not usually been seen in man or animals, their relevance to man is questionable.

Animals↗
PubMed1997

Identification of historical lead sources in roof dusts and recent lake sediments from an industrialized area: indications from lead isotopes.

X-ray fluorescence and stable lead (Pb) isotopic analyses have been undertaken on dusts, known from microscopic investigation to contain significant quantities of industrially- and urban-derived particulate matter, present in the roof cavities of houses in the Illawarra region (N.S.W., Australia), with the objective of examining the historic record of Pb pollution. All investigated houses contained in excess of 250 micrograms g-1 Pb, with dwellings close to a copper smelter, in a large industrial complex including a major steelworks, containing higher (> 2500 micrograms g-1) Pb concentrations. The isotopic composition in the dusts, expressed here as 206Pb/204Pb, is relatively constant at 17.0, irrespective of dwelling age or distance from the industrial complex. Contamination of the dusts by Pb sourced from paint cannot explain the isotopic uniformity of the dust samples. Isotopic modelling indicates that the dusts contain Pb derived from the copper smelter, gasoline-air Pb and a minor contribution from coal-utilising sources. Lead loading was also investigated in the adjacent lagoon, which acts as a natural sink for particulate matter in the Illawarra region. Isotopic data and modelling indicate that one natural and four anthropogenic sources contribute to the Pb burden of this lagoon. The natural source consists of Permian rocks cropping out in the catchment area which have a 206Pb/204Pb of approximately 18.7. The suggested anthropogenic sources are an old disbanded base-metal (Pb) smelter (206Pb/204Pb approximately 16.2-16.3), the copper smelter (206Pb/204Pb approximately 17.9), gasoline-air derived Pb (206Pb/204Pb approximately 16.4-16.5) and industries utilising coal, for example the recently closed thermal coal-fired power station (206Pb/204Pb approximately 18.9). The relative contributions of the base-metal (mainly lead) smelter and gasoline-air Pb in the sediment can only be partly assessed due to the isotopic similarity of these sources. Likewise the natural background and coal source (e.g. power station) contributions can only be estimated from historical data. Age estimations for sediment cores, using 137Cs, provide some control on these assessments. Near surface sediments in the lagoon have a relatively constant 206Pb/204Pb of 17.6-17.7, irrespective of sample location. Isotopic calculations, together with records of particulate matter pollution emissions, indicate a link between the Pb in roof dusts (206Pb/204Pb approximately 17.0) and Pb contamination of the near surface (upper 20 cm) lagoonal sediments via a homogeneous, non-unique source of lead whose isotopic composition closely matches that of the dusts. Over the last 5 decades, atmospheric fallout of Pb-bearing particulate matter appears to have been the dominant pathway for addition of Pb to the lagoon and dwellings in the Illawarra region.

Australia↗
PubMed1998

Lead poisoning of children in Africa, IV: Exposure to dust lead in primary schools in south-central Durban, South Africa.

Twenty primary schools in three areas (Merebank, Austerville and Bluff) of south-central Durban, South Africa were studied to determine if dust lead loading (microgram/m2) is high enough to constitute a risk to children. Atmospheric lead fallout rates in the schools varied from 9 to 264 micrograms/m2 per day and were higher than the values now being reported in the developed countries. Mean outside dust lead loadings were 425 micrograms/m2 in Merebank, 771 micrograms/m2 in Austerville and 1174 micrograms/m2 in Bluff areas, and 25% of the primary schools were found to have mean outside dust lead loading above 1000 micrograms/m2. Mean dust lead loadings inside classrooms were 2-4 times lower than the outside rates, and showed no significant difference among schools in the three areas. Dust lead loadings were correlated with lead concentrations in dust but not with distance from the highway or with atmospheric lead deposition rates. Exposure to dust lead in classrooms and in the school yard is considered to be an important contributor to blood lead levels in, and hence a potential health hazard to, children in many urban areas of the city.

Africa↗
PubMed1996

False-positive skin prick test responses to commercially available dog dander extracts caused by contamination with house dust mite (Dermatophagoides pteronyssinus) allergens.

BACKGROUND: In an outpatient population, a high frequency of positive skin prick test responses to dog dander was found in the absence of detectable IgE to dog dander in the RAST. The majority of these patients were sensitized to house dust mites (Dermatophagoides pteronyssinus) and had no obvious dog-related allergic symptoms. These findings prompted us to investigate whether dog dander skin test preparations are contaminated with house dust mite allergens in amounts sufficient to cause false-positive skin prick test responses in patients sensitized to house dust mites. METHODS: Antigen detection assays with monoclonal and polyclonal antibodies were used to determine concentrations of the major allergen Can f 1 from dog dander and the major allergens Der p 1 and Der p 2 from house dust mites in five commercially available dog dander skin prick test preparations (A to E). RESULTS: Can f 1 concentrations varied for the different extracts (A: 170 micrograms/ml, B: 11.1 micrograms/ml, C: 13.3 micrograms/ml, D: 3.8 micrograms/ml, and E: 59.4 micrograms/ml). Der p 1 was detectable in all extracts (A: 33.4 ng/ml, B:5.1 ng/ml, C:29.6 ng/ml, D: 0.4 ng/ml, and E: 1.9 ng/ml), and Der p 2 was detectable in some of the commercially available dog dander skin prick test preparations tested (A: 31.3 ng/ml, B: 3.0 ng/ml, and C: 7.5 ng/ml). The median house dust mite threshold in the skin prick test was found to be 5.8 ng/ml, of Der p 1 (range, 3.5 to 20.8 ng/ml) in nine patients tested. CONCLUSION: Contamination of commercially available dog dander skin prick test preparations with the major allergens (Der p 1 and Der p 2) of the house dust mite (D. pteronyssinus) was demonstrated. These contaminations cause false-positive responses to skin prick tests with dog dander in patients sensitized to house dust mite.

Allergens↗
PubMed1985

Short term in vivo method for prediction of the fibrogenic effect of different mineral dusts.

The effect of intratracheal introduction of different metal and mineral dusts and the change in activity of pulmonary acidic phosphatase have been studied as a function of time (72 h, 2 weeks, 1, 12, 20 months). The activity and localization of acid phosphatase were compared with the degree of pulmonary damage caused by dusts. The degree of fibrosis was determined on the basis of the composition of cells and fibres, according to Belt and King's classification. Due to the membrane-damaging effect of DQ 12 silica and mixed dusts (enargite and porphyry rock dusts) an increase in acid phosphatase activity of macrophages could be observed at the end of the first month. At the same time non-fibrogenic or only mild fibrogenic dusts (bentonite, corundum, scarnic rock dust) caused a decrease or disappearance of tissue acid phosphatase activity. It has been stated that there is a very close correlation between the change in pulmonary acidic phosphatase activity and the progression of pulmonary fibrosis due to exposure to mineral dusts. The above investigations have been most useful in predicting the subsequent effect of rock patterns, emphasizing at the same time the importance of in vivo long term experiments.

Acid Phosphatase↗
PubMed1998

Low dose effects of fibrous and non-fibrous mineral dusts on the proliferation of mammalian cells in vitro.

This report presents preliminary results of studies on the growth stimulating properties of mineral dusts. The studies were carried out with Syrian hamster diploid embryonic fibroblasts and Chinese hamster fibroblasts, line B14F28. Toxicity testing was done by the determination of the plating efficiency as a measure of growth and viability. The following dusts were used: the modified UICC asbestos fibres amosite, (AFF) crocidolite (KFF), chrysotile (KFF): in addition glass fibre (GFF), corundum and quartz DQ12. Concentrations > 2 microg/cm2 of AFF, CFF, KFF and GFF depressed the plating efficiency of B14F28 cells in a dose-dependent manner, but all of the fibres and corundum increased colony sizes at concentrations of 0.16-0.33 microg/cm2, in the case of corundum, AFF, KFF, and CFF also at up to 0.66 or 1.32 microg/cm2. DQ12 did not enhance colony growth. The stimulation of proliferation could be demonstrated both in terms of colony size (diameter) and cell numbers. The factor(s) responsible for proliferation stimulation reside in the supernatant, since the medium of dust-treated cell cultures was able to stimulate colony growth after removal of the dusts by filtration. The results indicate the induction of growth factors (cytokines) by low concentrations of the mineral dusts. Experiments concerning the effect of dusts on embryonic golden hamster fibroblasts yielded similar results. The plating efficiency was inhibited by concentrations of GFF and CFF > 0.25 microg cm2 and by AFF, KFF, GFF and corundum at concentrations > 5 microg/cm2, but colony counts were significantly increased by AFF, KFF and corundum at concentrations of 0.25-3 microg/cm2. This biological reaction which was observed in different cell types appears to be especially relevant in the context of environmental exposure where low dust concentrations prevail.

Animals↗
PubMed2003

Organic dust activates NF-kappaB in lung epithelial cells.

Exposure in swine confinement facilities induces airway inflammation in healthy subjects. The aim of the present study was to elucidate the role of nuclear factor (NF)-kappaB in the inflammatory response induced by organic dust. A human lung epithelial carcinoma cell line (A549) was transfected with reporter genes of the human IL-6 promoter or the NF-kappaB binding site fused to the luciferase reporter gene and stimulated with dust from a swine confinement building. Cytokine release in cell culture supernatants and luciferase activity was measured. The dust-induced the activities of the IL-6 promoter reporter gene and the NF-kappaB reporter gene in parallel with an increase in IL-6 and IL-8 release. The addition of pyrrolidinedithiocarbamate, a chemical NF-kappaB blocking agent, inhibited IL-6 and IL-8 secretion as well as the NF-kappaB reporter gene activity. Increasing the amount of IkappaB alpha led to inhibition of organic dust-induced IL-6 promoter and NF-kappaB reporter gene activities. Fluticasone inhibited the organic dust-induced NF-kappaB activation and IL-6 and IL-8 secretion. Finally, swine dust incubation of A549 cells resulted in a NF-kappaB DNA binding, which is composed of the NF-kappaB1 and RelA proteins. In conclusion, by interference at various levels we have shown that NF-kappaB plays a key role in the inflammatory response to organic dust.

Androstadienes↗
PubMed2001

Allergenic materials in the house dust of allergy clinic patients.

INTRODUCTION: Environmental agents including animal, fungal, tree, and weed antigens are known to cause allergic rhinitis and asthma. The following study was performed to measure the antigen concentration of several of these in house dust of children seen in an allergy clinic. Comparisons are made between household allergen levels of children seen for asthma and children seen for other reasons. METHODS: Dust samples were solicited from patients in a pediatric allergy specialty clinic and other individuals associated with the clinic. Persons submitting dust were asked to complete a questionnaire describing their house. Samples were extracted, centrifuged, and filtered for sterility. Samples were stored in 50% glycerol at -20 degrees C. Specific antigens for Alternaria, Cladosporium, Aspergillus, Candida, Dermatophagoides farinae, cat, dog, oak, fescue, ragweed, plantain, and cockroach were measured using inhibition assays developed with whole antigen extract. Allergens Der p1, Der f 1, Alt a 1, and Alt a 70 kD were measured using double monoclonal antibody assays. RESULTS: Significant concentrations of whole antigen from cat, dog, oak, Alternaria, and Cladosporium were detected. Between 0.1 and 18 microg of Der f1 and Der p1 per gram of dust were also measured. Alt a 1 and Alt a 70 kD levels varied between 3.0 and 1000 U/g of dust. Significant positive correlations were observed in levels of dust mite and Alternaria allergen for patients with an evaluation of asthma. CONCLUSIONS: We found measurable levels of fungal antigens (Alternaria, Cladosporium), mite antigens, and animal antigens (dog and cat) in the majority of dust samples in this self-selected set of allergy clinic patients. Specific allergens Alt a 1, Alt a 70kD, and Der p 1 were significantly higher in the homes of asthmatic patients when compared with patients seen for reasons other than asthma. These studies support the hypothesis that fungal allergen exposure is an important component in the pathogenesis of the clinical condition known as asthma.

Air Pollution, Indoor↗
PubMed1997

Identification of IgE-binding components in occupational asthma caused by corn dust.

BACKGROUND: There have been some reports of corn dust-induced occupational asthma that suggest nonimmunologic mechanism. In this paper, we present a case of occupational asthma and rhinitis caused by corn dust where bronchoconstriction was induced by an IgE-mediated reaction. METHODS AND RESULTS: The patient had positive responses to weed and grass pollens as well as corn dust extracts. The bronchoprovocation test elicited an early asthmatic response to corn dust extracts. Serum-specific IgE and IgG4 antibodies to corn dust extracts were detected by enzyme-linked immunosorbent assay (ELISA). In order to identify further the allergenic component of the extracts, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotting studies were performed. Ten IgE-binding components (9 to 140 kD) were detected within the corn dust extracts. CONCLUSION: These results suggest that inhalation of corn dust can cause IgE-mediated bronchoconstriction in an exposed worker.

Adult↗
PubMed1996

Microflora and acarofauna of bed dust from homes in Upper Silesia, Poland.

BACKGROUND: Pyroglyphid mites are considered a major cause of house dust allergy. The occurrence and possible pathogenic role of other biologic components of house dust, in particular bacteria, has received less attention. OBJECTIVE: The aim of this study was to examine bacteria present in the samples of house dust from beds, in comparison to fungi and mites recovery. METHODS: Samples of bed dust were collected from 40 homes in Upper Silesia (Poland). Of these, 19 came from the homes of people with asthma caused by house dust and 21 from the homes of people without allergy. The concentrations of bacteria, fungi, mites, and endotoxin and species composition of microflora and acarofauna were determined. RESULTS: The overall mean concentrations of mesophilic bacteria, thermophilic bacteria and fungi, including yeasts, were, respectively, 1.6 x 10(6), 1.7 x 10(3), and 1.6 x 10(4) CFU/g. Samples contained an average of 8.4 mites/g and the ten samples assayed for bacterial endotoxin averaged 80.4 ng/mg. A total of 55 species of bacteria, 40 of fungi and 13 of mites were found. Gram-positive cocci (mostly Staphylococcus spp.) were the predominant mesophilic bacteria, followed by corynebacteria and Bacillus spp. Thermophilic bacteria were represented only by actinomycetes, with Thermoactinomyces vulgaris predominant. The most numerous fungi were Penicillium spp. and Aspergillus spp. followed by yeasts. The most abundant mites were Dermatophagoides spp. which formed > 85% of the total count. There were no significant differences between the homes of allergic and nonallergic people in the concentrations of total bacteria, fungi, and mites. Bacillus, Aspergillus and total filamentous fungi (molds), but not yeasts, were significantly more numerous in the homes of people with asthma caused by house dust. CONCLUSION: The results suggest that some species of bacteria and filamentous fungi should be considered potential causes of house dust allergy.

Animals↗
PubMedDate not supplied

Factors that influence the formation and stability of hydrated ferrous sulfate in coal dusts. Possible relation to the emphysema of coal miners.

Epidemiological studies have shown that a causal relationship may exist between coal dust exposure and emphysema in coal miners. Emphysema can be considered as one of the human pathologies associated with oxidative stress, resulting from oxidant-induced alpha 1-antitrypsin (alpha 1-AT) inactivation and uncontrolled proteolysis of lung tissue. We have previously reported that certain coal dusts contained hydrated ferrous sulfate (FeSO4) that inactivated alpha 1-AT [Huang et al. (1993) Chem. Res. Toxicol. 6, 452-458]. In the present study, we have shown that the FeSO4 originated from oxidation of pyrite (FeS2), which is a typical contaminant of coal dusts. The relative humidity and microenvironment around individual pyrite particles influence the formation of FeSO4 in the coal. However, the subsequent human exposure to coal dust containing FeSO4 depends on the stability of the formed FeSO4. We found that pH played the most important role in stabilizing the FeSO4, such that a final pH < 4.5 after oxidation of pyrite stabilized FeSO4, whereas at high pH the conversion of reactive Fe2+ to Fe3+ was immediate. Sulfuric acid (H2SO4), which is also produced by the oxidation of pyrite, can lower the pH, but it can also be neutralized by other minerals in coal dusts, such as calcite (CaCO3). The stability of FeSO4 in coal dust can also be influenced by the length of exposure to air. Our studies demonstrated that coal samples differed in their capacity to stabilize FeSO4. This current study strengthens our previous reported hypothesis that emphysema, which occurs irregularly in coal miners, could be directly related to exposure to coal dust containing FeSO4.

Coal↗
PubMed1999

Low-temperature crystallization of silicate dust in circumstellar disks.

Silicate dust in the interstellar medium is observed to be amorphous, yet silicate dust in comets and interplanetary dust particles is sometimes partially crystalline. The dust in disks that are thought to be forming planets around some young stars also appears to be partially crystalline. These observations suggest that as the dust goes from the precursor clouds to a planetary system, it must undergo some processing, but the nature and extent of this processing remain unknown. Here we report observations of highly crystalline silicate dust in the disks surrounding binary red-giant stars. The dust was created in amorphous form in the outer atmospheres of the red giants, and therefore must be processed in the disks to become crystalline. The temperatures in these disks are too low for the grains to anneal; therefore, some low-temperature process must be responsible. As the physical properties of the disks around young stars and red giants are similar, our results suggest that low-temperature crystallization of silicate grains also can occur in protoplanetary systems.

Cold Temperature↗
PubMed1989

Adjuvant activity of respirable iron ore dust.

The influence of the respirable fraction of an iron ore dust on immunity has been studied. The iron ore dust enhanced the specific immune responses to both sheep erythrocytes and a bacterial lipopolysaccharide. The IgG class of antibody was enhanced significantly and its pattern of enhancement suggested that the iron ore dust was functioning as an adjuvant. In studies to test this possibility further, the pattern of antibody development over both the primary and the secondary immune response to sheep erythrocytes closely matched that of Freund's incomplete adjuvant and low levels of silica. The splenocytes, from animals that had received iron ore dust for various lengths of time, exhibited enhanced polyclonal mitogenic activity for phytohaemagglutinin, in vitro. These properties were seen when the dust had been inhaled or injected into the pleural cavity and persisted for over 1 year following the injection or inhalation of the dust. Attempts to show the induction of interleukin-1 by the macrophages from mice implanted intraperitoneally with the iron ore dust were unsuccessful.

Adjuvants, Immunologic↗
PubMedDate not supplied

Quantification of Stachybotrys chartarum conidia in indoor dust using real time, fluorescent probe-based detection of PCR products.

Analyses of fungal spores or conidia in indoor dust samples can be useful for determining the contamination status of building interiors and in signaling instances where potentially harmful exposures of building occupants to these organisms may exist. A recently developed method for the quantification of Stachybotrys chartarum conidia, using real-time, fluorescence probe--based detection of PCR products (TaqMan system) was employed to analyze indoor dust samples for this toxigenic fungal species. Dust samples ofup to 10 mg were found to be amenable to DNA extraction and analysis. Quantitative estimates of S. chartarum conidia in composite dust samples, containing a four-log range of these cells, were within 25 -- 104% of the expected quantities in 95% of analyses performed by the method. Calibrator samples containing known numbers of S. chartarum conidia were used as standards for quantification. Conidia of an arbitrarily selected strain of Geotrichum candidum were added in equal numbers to both dust and calibrator samples before DNA extraction. Partial corrections for reductions in overall DNA yields from the dust samples compared to the calibrator samples were obtained by comparative analyses of rDNA sequence yields from these reference conidia in the two types of samples. Dust samples from two contaminated homes were determined to contain greater than 10(3) S. chartarum conidia per milligram in collection areas near the sites of contamination and greater than 10(2) conidia per milligram in several areas removed from these sites in analyses performed by the method. These measurements were within the predicted range of agreement with results obtained by direct microscopic enumeration of presumptive Stachybotrys conidia in the same samples.

Air Pollution, Indoor↗