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1.
Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide (LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids (3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS (LPS) results between laboratories for all dust types (p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs (p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes.  相似文献   

2.
Studies on personal dust and endotoxin concentrations among animal farmers have been either small or limited to a few sectors in their investigations. The present study aimed to provide comparable information on the levels and variability of exposure to personal dust and endotoxin in different types of animal farmers. 507 personal inhalable dust samples were collected from 327 farmers employed in 54 pig, 26 dairy, 3 poultry, and 3 mink farms in Denmark. Measurements in pig and dairy farmers were full-shift and performed during summer and winter, while poultry and mink farmers were monitored during 4 well-defined production stages. The collected samples were measured for dust gravimetrically and analyzed for endotoxin by the Limulus amebocyte lysate assay. Simple statistics and random-effect analysis were used to describe the levels and the variability in measured dust and endotoxin exposure concentrations. Measured inhalable dust levels had an overall geometric mean of 2.5 mg m(-3) (range 相似文献   

3.
4.
Endotoxin is a toxic, pro-inflammatory compound that has been detected in indoor air and dust in homes and occupational settings, and also in outdoor air. Data on the outdoor sampling of endotoxin are limited. Currently, little is known about the seasonal variation and influence of temperature on outdoor endotoxin levels. In the present study, we report endotoxin levels in fine fraction particulate matter with a 50% aerodynamic cutoff diameter of 2.5 microm (PM2.5) and describe the seasonal variation of endotoxin in Munich, Germany. In 1999-2000, PM2.5 was collected at forty outdoor monitoring sites across Munich. Approximately four samples were collected at each site for a total of 158 samples. Endotoxin concentrations in the PM2.5 samples were determined using the kinetic chromogenic Limulus Amebocyte Lysate (LAL) assay. The geometric mean endotoxin concentration was 1.07 EU mg PM2.5(-1) (95% C.I.: 0.915-1.251) or 0.015 EU m(-3) of sampled air (95% C.I.: 0.013-0.018). Munich endotoxin levels were significantly related to ambient temperature (p < 0.0001) and percent relative humidity (p < 0.0001). Sampling periods with higher average temperatures yielded higher levels of endotoxin in PM2.5 (r = 0.641), whereas decreases in percent relative humidity were associated with increased endotoxin levels in PM2.5 (r = -0.388). Endotoxin levels were significantly higher during the warmer seasons of spring [means ratio (MR): 2.5-2.7] and summer (MR: 2.1-3.0) than during winter. Although temperature and relative humidity do not explain all of the variability in endotoxin levels, their effects were significant in our data set. Temperature effects and seasonal variation of endotoxin should be considered in future studies of outdoor endotoxin.  相似文献   

5.
Traditional methods of assessing fungal exposure have been confounded by a number of limiting variables. The recent utilization of molecular methods such as internal transcribed spacer (ITS) sequencing of ribosomal RNA genes has provided improved insight into the diversity of fungal bioaerosols in indoor, outdoor and occupational environments. However, ITS analyses may also be confounded by a number of methodological limitations. In this study, we have optimized this technology for use in occupational or environmental studies. Three commonly used DNA extraction methodologies (UltraClean Soil kit, High Pure PCR Template kit, and EluQuik/DNeasy kit) were compared in terms of sensitivity and susceptibility to PCR inhibitors in dust for three common fungal bioaerosols, Aspergillus versicolor, Rhizopus microsporus and Wallemia sebi. Environmental dust samples were then studied using each extraction methodology and results were compared to viable culture data. The extraction methods differed in terms of their ability to efficiently extract DNA from particular species of fungi (e.g. Aspergillus versicolor). In addition, the ability to remove PCR inhibitors from dust samples was most effective using the soil DNA extraction kit. The species composition varied greatly between ITS clone libraries generated with the different DNA extraction kits. However, compared to viable culture data, ITS clone libraries included additional fungal species that are incapable of growth on solid culture medium. Collectively, our data indicated that DNA extraction methodologies used in ITS sequencing studies of occupational or environmental dust samples can greatly influence the fungal species that are detected.  相似文献   

6.
Gas chromatography-mass spectrometry (GC-MS) using a quadrupole instrument and GC-tandem MS (GC-MS-MS) using an ion trap instrument were applied to determine 3-hydroxy fatty acids (3-OH FAs) with 10-18 carbon chain lengths, specific components of the endotoxin (lipopolysaccharide, LPS) of Gram-negative bacteria, in 30 house dust samples. The two methods provided similar detection sensitivity for methyl ester/trimethylsilyl derivatives of the 3-OH FAs and allowed these acids to be distinguished from co-eluting 2-OH FA derivatives. The correlation coefficients between endotoxin activity (Limulus test) and the combined amounts of 3-OH C10, 3-OH C12, and 3-OH C14 were 0.60 and 0.61 when using GC-MS and GC-MS-MS, respectively. The superior selectivity of GC-MS-MS was illustrated in analyses of sub-milligram amounts of dust, where the chromatograms achieved by GC-MS were difficult to interpret due to a high background and several closely eluting compounds. GC-MS-MS is therefore preferable to GC-MS for determining 3-OH FAs in minute (sub-milligram) amounts of dust.  相似文献   

7.
Elevated concentrations of endotoxin in indoor air due to cigarette smoking   总被引:1,自引:0,他引:1  
Exposure to environmental tobacco smoke (ETS) is an important worldwide public health issue. The present study demonstrates that cigarette smoke can be a major source of endotoxin (lipopolysaccharide, LPS) in indoor environments. Gas-chromatography/mass-spectrometry was used to determine 3-hydroxy fatty acids as markers of endotoxin in air-borne house dust in homes of smokers and non-smokers. Air concentrations of endotoxin were 4-63 times higher in rooms of smoking students than in identical rooms of non-smoking students. The fact that cigarette smoke contains large amounts of endotoxin may partly explain the high prevalence of respiratory disorders among smokers and may also draw attention to a hitherto neglected risk factor of ETS.  相似文献   

8.
In this study, air samples were taken using a BioSampler and gelatin filters from six sites in Beijing: office, hospital, student dormitory, train station, subway, and a commercial street. Dust samples were also collected using a surface sampler from the same environments. Limulus amoebocyte lysate (LAL) and Glucatell assays were used to quantify sample endotoxin and (1,3)- ${\rm{\beta}} $ -d-glucan concentration levels, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to measure the dust mite allergens (Der p 1 and Der f 1). Ultrafine particle and lead concentrations in these sampling sites were also measured using P-Trak and atomic absorption spectrometer, respectively. Analysis of variance (ANOVA) and linear regression analysis were used to analyze the concentration data. Higher culturable bacteria (12,639 CFU/m3) and fungi (1,806 CFU/m3) concentrations were observed for the train station and the subway system, respectively. For the rest of sampling sites, their concentrations were comparable to those found in western countries, ranging from 990 to 2,276 CFU/m3 for bacteria, and from 119 to 269 CFU/m3 for fungi. ANOVA analysis indicated that there were statistically significant differences between the culturable bacterial and fungal concentration levels obtained for different sites (p value = 0.0001 and 0.0047). As for dust allergens, endotoxin, and (1,3)-β -d-glucan, their concentrations also seemed to be comparable to those found in the developed countries. Airborne allergen concentrations ranged from 16 to 68 ng/m3. The dust-borne allergen concentration was observed to range from 0.063 to 0.327 ng/mg. As for endotoxin, the highest airborne concentration of 25.24 ng/m3 was observed for the commercial street, and others ranged from 0.0427 to 0.1259 ng/m3. And dust-borne endotoxin concentration ranged from 58.83 to 6,427.4 ng/mg. For (1,3)-β -d-glucan, the airborne concentration ranged from 0.02 to 1.2 ng/m3. Linear regression analyses showed that there existed poor correlations between those in airborne and dust-borne states (R2?=?0.002~0.43). In our study, the lowest ultrafine particle concentration about 5,203 pt/cm3 was observed in office and the highest was observed at the train station, up to 32,783 pt/cm3. Lead concentration was shown to range from 80 to 170 ng/mg with the highest also observed at the train station. The information provided in this work can be used to learn the general situation of relevant health risks in Beijing. And the results here suggested that when characterizing exposure both airborne and dust-borne as well as the environments should be considered.  相似文献   

9.
The aim of this study was to assess the associations between airborne and dustborne microbial contaminants (endotoxin and β-D-glucan) and estimate the effects of home characteristics on exposure levels of these microbial contaminants. Endotoxin and β-D-glucan concentrations in airborne inhalable particles, airborne PM1 (<1 μm) and vacuumed dust from 184 residential homes were determined using specific Limulus amebocyte assays. Home characteristics were recorded by visual inspection and questionnaires. Linear regression and correlation analyses were performed. Inhalable endotoxin correlated with dust endotoxin (r = 0.34, p < 0.001) and PM1 endotoxin (r = 0.33, p < 0.001). Inhalable β-D-glucan correlated with dust β-D-glucan (r = 0.18, p < 0.01), but not with PM1 β-D-glucan. Significant correlation was also found between PM1 and dust concentrations for endotoxin (r = 0.26, p < 0.001), but not for β-D-glucan. Multivariate regression analyses showed only one significant association between airborne contaminants and environmental characteristics: inhalable β-D-glucan was positively associated with relative humidity with an effect size (change in the dependent variable corresponding to a unit increase in the independent variable) of 2.32 and p < 0.05. In contrast, several associations were found between dust concentrations and environmental characteristics. Dust endotoxin was positively associated with temperature (2.87, p < 0.01) and number of inhabitants (2.76, p < 0.01), whereas dust β-D-glucan was inversely associated with the presence of dogs (-2.24, p < 0.05) and carpet (-3.05, p < 0.01) in the home. In conclusion, dustborne contaminants were more strongly affected by home characteristics than airborne contaminants. Furthermore, even though statistically significant, the correlations between airborne and dustborne contaminants were weak. This indicates that airborne concentrations cannot be reliably predicted based on dustborne concentrations.  相似文献   

10.
A series of perfluorinated compounds (PFCs) including perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) have been recently measured in a variety of environmental samples and biological matrices. In order to better understand the human exposure routes of these chemicals, levels of PFOS, PFOA, perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHS) and perfluorooctane sulfonamide (PFOSA) in house dust samples were investigated. The data revealed a correlation between the concentrations of PFCs and the percentage of carpeting in the house; older houses tended to have less carpeting, hence lower levels of these perfluorinated compounds in their dust.  相似文献   

11.
Airborne particulates have a major influence on the quality of indoor environments, which in turn affects human health. Both allergens and proteases are major etiological factors in allergic disease. Thus, the monitoring of environmental protease particulates is very important for the management of allergic disease. Protease-monitoring systems have been introduced in industry, but have not been applied to domestic settings, probably because of the high cost and many complex steps involved in these systems. In this study, we developed an indoor protease-monitoring system which can easily detect protease particulates without need for pre-treatment of dust samples or complex measuring instruments such as fluorescent plate reader. As a protease substrate, gelatin thin film (GTF) was adopted to evaluate small amounts of house dust particulates. The principle of this assay is based on the hydrolysis of GTF. Protease molecules spread from a particulate to GTF can hydrolyze the gelatin, thereby producing a spot on the GTF. This hydrolyzed spot visualized by staining was much larger than the particulate size, making visible to naked eye. The GTF method facilitated the observation of a subtle increase in protease particulates fallen on the GTF after the turnover of a used bed-quilt. The newly developed GTF method seems to be a suitable tool for protease monitoring at home.  相似文献   

12.
Muramic acid (MA) is a unique amino sugar that is a constituent of the peptidoglycan (PG) present in prokaryotic cell walls. MA can serve as a marker for quantifying bacterial load, e.g. in indoor environments, by using gas chromatography-tandem mass spectrometry (GC-MS-MS). We demonstrated recently that the methyl ester O-methyl acetate (MMA) derivative can be used to detect MA in house dust by ion-trap GC-MS-MS. However, since the MMA derivative is not formed from free MA quantification was not optimal. Here we report 1) significant improvements in sample preparation of the MMA derivative and 2) an evaluation of the performance of derivative, using for comparison the alditol acetate derivative, the gold standard in quantitative trace analysis of MA in complex matrices. The MMA derivative was analysed using an MS instrument with internal ionization and the alditol acetate derivative was analysed using an instrument with external ionization. 13C-labelled cyanobacteria, containing MA in their PG, were used as the internal standard. A linear relationship was found between the two methods in studies on 27 parallel samples of airborne dust from school classes collected on filters. Although the analytical sensitivity of the MMA derivatives was somewhat slightly lower than of the alditol acetate derivative, this may be due to differences in yield of derivative, sample clean-up efficiency, or different performance of the GC columns or MS instruments. However preparation of the MMA derivative is quick and compatible with preparation of methyl esters of 3-hydroxy fatty acids (used as markers of Gram negative endotoxin) allowing the levels of both markers to be determined in the same dust sample. In conclusion, the MMA procedure can be used to determine MA in environmental samples with good reproducibility provided the concentration of the 13C-labelled MA internal standard in the cyanobacteria is first determined with an alternative method.  相似文献   

13.
Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml(-1) in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml(-1) in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml(-1) of four real-life dust samples at p-value >0.05 using 2 x 10(4) spores for conventional PCR and 2 x 10(5) spores for real-time PCR. However, samples containing >0.2 mg ml(-1) real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments.  相似文献   

14.
This study was carried out to measure and compare the concentration of bacterial endotoxin in a variety of samples from drinking tap and bottled water available in Kuwait by using the Limulus Amoebocyte lysate test. A total of 29 samples were tested. Samples were collected from a variety of locations throughout the six governorates of Kuwait and 23 brands of local and imported bottled water samples were collected from the local market. The concentration of bacterial endotoxin was measured by using the standard Limulus Amoebocyte lysate test, gel clot method. This study showed that measured endotoxin concentrations in tap drinking water varied from 2.4 to 33.8?EU/ml with the average endotoxin concentration of 14.2?EU/ml. While the results of endotoxin concentrations in the bottled water were <0.03 to 20.1?EU/ml with an average of 1.96?EU/ml. The average concentration of endotoxin in bottled water is 13.5 % of the average concentration of endotoxin in tap drinking water. This experimental investigation has proved that drinking bottled water has less endotoxin as compared to tap water in Kuwait. It is also demonstrated that the endotoxin concentration did not exceed the acceptable level in drinking tap water.  相似文献   

15.
Symptoms such as shortness of breath and cough have been noted in woodworking facilities even where wood dust itself is well-controlled. Suspicion has fallen on other possible contaminants in the workplace atmosphere, including bacterial endotoxin. A few studies have indicated potentially high endotoxin exposure with exposure to fresh wood in sawmills and in the production of fiberboard and chipboard, but fewer studies have been carried out on exposure to endotoxin in dry wood work, for example in joineries. A study of the endotoxin content of airborne wood dust samples from US woodworking facilities is presented, from the re-analysis of samples which previously had been taken to establish mass collection relationships between the IOM sampler, the closed-face 37 mm plastic cassette (CFC) sampler and the Button sampler. Endotoxin was strongly correlated with total dust, but the endotoxin content of a few fresh wood samples was found to be up to ten times higher per unit of wood dust than for dried-wood samples, and this difference was significant. No long-term time-weighted average sample exceeded the recommended limit value of 50 EU m(-3) (EU, endotoxin units)used in the Netherlands, although a number of the IOM samples came close (seven samples or 44% exceeded 20 EU m(-3)) and one short-term (48 minute) sample registered a high value of 73 EU m(-3). The geometric mean concentration from the IOM samples (11 EU m(-3)) is within the range of geometric means found from Australian joineries (3.7-60, combined: 24 EU m(-3)). In contrast, the corresponding values from the CFC (3.6 EU m(-3)), and the Button sampler (2.1 EU m(-3)) were much lower and no samples exceeded 20 EU m(-3). Endotoxin is likely only to be a significant problem in working with dried woods when associated with very high dust levels, where the wood dust itself is likely to be a cause for concern. The results from the few samples in this study where fresh wood was being worked were similar to results from other studies involving fresh woods. The agreement between these studies is encouraging given the difficulties of endotoxin analysis and the wide variation often expected between different laboratories.  相似文献   

16.
Allergy to rodent proteins is common among laboratory animal workers. Sensitive methods to measure exposure to these allergens have been developed. These assays are, however, expensive, time-consuming, and require a laboratory facility and methodological expertise. A simple method to screen for allergen spread, or to test whether hygiene standards are maintained, would be useful. Lateral flow immunoassays (LFIAs) are especially suited for field settings; the tests are simple and results are visible within minutes. LFIAs were developed for detection of the rodent urinary allergens Mus m 1 and Rat n 1. Pilot studies were performed in animal facilities in three countries using both extracts from airborne dust samples and samples collected by wiping surfaces. For comparison and determination of sensitivity, the concentrations of rodent urinary allergens in the samples were also measured using enzyme immunoassays (EIAs). The LFIAs for rat and mouse urinary allergens had a detection limit of 31 pg allergen per mL in a buffer system with purified allergen standards. Results of environmental dust extracts tested in LFIAs correlated well with levels obtained using EIAs. Spread of rodent allergens, or non-adherence to hygiene around laboratory animal facilities, may aggravate rodent allergy. Using a simple, sensitive one-step assay, allergens can be detected to prevent allergen exposure. The results reveal that the rapid assays are suited for on-site demonstration of exposure to rodent allergens, and thus, useful in occupational hygiene practice.  相似文献   

17.
House dust has been identified as a major exposure medium for lead (Pb) in children. High levels of Pb in soil and house dust have been recorded at the Bunker Hill Superfund Site (BHSS) in northern Idaho, an historic mining and smelting district. Soil and dust remediation at the site was required; however, regional background soil and dust Pb levels had not been well characterized. The objective of this survey was to determine background house dust Pb levels and to compare those levels with concentrations, and dust and Pb loading rates measured at the BHSS. Soil and house dust samples were collected in five towns demographically similar to the BHSS but unaffected by the mining industry. The background concentrations and loading rates were significantly lower than those observed at the site. House age was a significant factor affecting background soil and house dust Pb concentrations and loading rates.  相似文献   

18.
Recent studies have demonstrated the utility of ultrasonic extraction (UE), followed by portable anodic stripping voltammetry (ASV), for the on-site determination of lead in environmental and industrial hygiene samples. The aim of this work was to conduct an interlaboratory evaluation of the UE-ASV procedure, with a goal of establishing estimates of method performance based on results from collaborative interlaboratory analysis. In this investigation, performance evaluation materials (PEMs) with characterized lead concentrations were used for interlaboratory testing of the UE-ASV procedure. The UE-ASV protocol examined has been promulgated in the form of two separate national voluntary consensus standards (one for UE and another for electroanalysis, which includes ASV). The PEMs consisted of characterized and homogenized paints, soils, and dusts (the last of which were spiked onto wipes meeting national voluntary consensus standard specifications), and air filter samples (mixed cellulose ester membrane) generated using characterized paints within an aerosol chamber. The lead concentrations within the PEMs were chosen so as to bracket pertinent action levels for lead in the various sample matrices. The interlaboratory evaluation was conducted so as to comply with an applicable national voluntary consensus standard that can be used to estimate the interlaboratory precision of a given analytical test method. Based on the analytical results reported by the participating laboratories, relative standard deviations (RSDs) for repeatability and reproducibility were computed for three different lead contents of the four PEMs. RSDs for repeatability were 0.019-0.100 for paints; 0.030-0.151 for soils; 0.085-0.134 for dust wipes; and 0.095-0.137 for air filters. RSDs for reproducibility were 0.127-0.213 for paints; 0.062-0.162 for soils; 0.085-0.134 for dust wipes; and 0.114-0.220 for air filters. With the exception of one of the air filter samples and one of the paint samples, the precision estimates were within the +/- 20% precision requirement specified in the US Environmental Protection Agency National Lead Laboratory Accreditation Program (NLLAP). The results of this investigation illustrate that the UE-ASV procedure is an effective method for the quantitative measurement of lead in the matrices evaluated in this study.  相似文献   

19.
All major opencast mining activities produce dust. The major operations that produce dust are drilling, blasting, loading, unloading, and transporting. Dust not only deteriorates the environmental air quality in and around the mining site but also creates serious health hazards. Therefore, assessment of dust levels that arise from various opencast mining operations is required to prevent and minimize the health risks. To achieve this objective, an opencast coal mining area was selected to generate site-specific emission data and collect respirable dust measurement samples. The study covered various mining activities in different locations including overburden loading, stock yard, coal loading, drilling, and coal handling plant. The dust levels were examined to assess miners' exposure to respirable dust in each of the opencast mining areas from 1994 to 2005. The data obtained from the dust measurement studies were evaluated by using analysis of variance (ANOVA) and the Tukey-Kramer procedure. The analyses were performed by using Minitab 14 statistical software. It was concluded that, drilling operations produce higher dust concentration levels and thus, drill operators may have higher incidence of respiratory disorders related to exposure to dust in their work environment.  相似文献   

20.
The aim of this work is to characterize the mineral phases present in the atmosphere at three locations in northern Oman. Samples of atmospheric particles were collected using a high volume sampler. Three indoor and one outdoor location were chosen in this investigation. Sampling locations included a residential house located nearby the cement plant, a residential house located nearby a refinery plant, and a residential house located at Al-Suwayq residential area. Indoor air was sampled from these three houses. Moreover, for the Al-Suwayq residential house, sampling was also taken outside the house for comparison. The dust samples were analysed for their microanalysis characterization and their mineral contents as well. The microanalysis enabled us to identify the metals present in the particles. Furthermore, the mineralogical analysis of the sample filters showed the presence of quartz as the principal phase inside the house of Al-Suwayq, whereas quartz, dolomite, and gypsum were common phases outside the house. In the residential house nearby the cement plant, it was found that calcite, quartz, dolomite and goethite were the principal phases whereas the particles collected from the house nearby the refinery composed primarily of dolomite and calcite. The airborne dust collected at the refinery and Al-Suwayq were probably sourced in the natural environment and mobilised by natural processes. However, at the cement factory the crushing and grinding of limestone during the industrial process has contributed significantly to the airborne dust load. Generally, the information obtained in this study will be invaluable as no data for the mineral content of atmospheric dust existed in the Oman.  相似文献   

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