Content of clinker and other materials in personal thoracic aerosol samples from cement plants estimated by scanning electron microscopy and energy-dispersive X-ray microanalysis.

OBJECTIVES: To estimate the composition and exposure to clinker and other specific components in personal thoracic dust samples of cement production workers. METHODS: A procedure for the classification of airborne particles in cement production plants was developed based on classification trees. For this purpose, the chemical compositions of 27,217 particles in 29 material samples (clinker, limestone, gypsum, clay, quartz, bauxite, iron source, coal fly ash, and coal) were determined automatically by scanning electron microscopy (SEM) and energy-dispersive X-ray microanalysis (EDX). The concentrations of the major elements in cement (calcium, aluminium, silicon, iron, and sulphur) were used for the classifications. The split criteria of the classification trees obtained in the material samples were used to classify 44,176 particles in 34 personal thoracic aerosol samples. The contents of clinker and other materials were estimated, and the clinker contents were analysed statistically for differences between job types and job tasks. RESULTS: Between 64% and 88% of the particles from material samples were classified as actual materials. The material types with variable composition (clay, coal fly ash, and coal) were classified with the lowest consistency (64% to 67%), while materials with a more limited compositional variation (clinker, gypsum, and quartz) were classified more consistently (76% to 85%). The arithmetic mean (AM) of the clinker content in personal samples was 62.1%, the median was 55.3%, and 95% confidence interval (CI) was 42.6% to 68.1%. No significant differences were observed between job types. However, the clinker content in samples when workers handled materials with high clinker content was significantly higher than when materials with lower clinker content were handled, 85% versus 65% (P = 0.02). The limestone content was AM 14.8%, median 13.2% (95% CI 5.5 to 20.9), whereas the other materials were present with relative abundances of median ≤ 6.4%. DISCUSSION: Automated particle analysis by SEM-EDX followed by classification tree analysis quantified clinker with fairly high consistency when evaluated together with raw materials that are expected to be airborne in cement production plants. The clinker proportions for job types were similar. Tasks a priori ranked by assumed clinker content were significantly different and according to expectations, which supports the validity of the chosen methodology. CONCLUSIONS: The composition of personal samples of mineral aerosols in the cement production industry could be estimated by automated single particle analysis with SEM-EDX and classification by a classification tree procedure. Clinker was the major component in the thoracic aerosol that cement production workers were exposed to. Differences between job types were relatively small and not significant. The clinker content from tasks was in agreement with assumptions.

A first account of the heterotrophic eukaryote Rabdiophrys Rainer from the fossil record and description of a new species from an ancient Eocene Arctic freshwater lake.

Rotosphaerids are unicellular, heterotrophic, eukayotic protists that have filopodia, an exterior covering consisting of highly ornamented siliceous scales, and are classified in the Rotosphaerida within the opistokont lineage. Given their appearance as relatively large spherical cells with protruding filopodia and a silica scale covering, they are often mistaken for centrohelid heliozoans. Even though these organisms are widely distributed in both marine and freshwater environments, many species are rarely reported, and none have been reported from the fossil record. We report extensive remains of a new species of Rabdiophrys, R. giraffensis, from an ancient waterbody that was situated near the Arctic Circle in northern Canada during the Eocene. The new species has both plate and spine scales that are similar in morphology, but significantly larger than its closest modern congeners, R. monopora and R. anulifera. The waterbody in which the new species grew and thrived is inferred to have been a moderately deep, circumneutral pond, with moderate concentrations of nutrients and dissolved humic material.

Kinetics and tissue distribution of bismuth, tin and lead after implantation of miniature shotgun alloy pellets in rats.

INTRODUCTION: Shotgun pellets containing bismuth (Bi) as substitute for lead (Pb) are increasingly being used due to environmental concerns. Information on toxicokinetics of Bi is lacking for the assessment of humans accidentally shot by Bi-containing shotgun alloy pellets. METHODS: Male Wistar rats were exposed to miniature alloy pellets containing Bi, tin (Sn) and minor amounts of Pb by implantation in muscle tissues of the hind legs. RESULTS: The concentrations of Bi in whole blood and urine increased up to 53 weeks after implantation. The highest concentrations of Sn in whole blood were observed three weeks after implantation, then declining to background levels 53 weeks after implantation. Lead in whole blood increased up to 13 weeks of exposure, and declined for the remaining observation period. Bismuth and Sn accumulated mainly in kidney, but also in liver, testicle and brain. Analytical field emission scanning electron microscopy of post-implant pellets showed depletion of Pb towards the pellet surface. Oxygen and chlorine accumulated in Sn rich lamellas in areas next to the pellet surface. The distribution of Bi remained visually unaffected as compared to pre-implant pellets. CONCLUSION: The concentration of Bi increased during the whole observation period in blood, urine, kidney, brain, testicle and liver. The decline in the concentrations of Pb and Sn in blood and urine after reaching the peak concentration may be related to alterations in the chemical composition and element distribution of the implanted alloy pellets.

Phase identification of individual crystalline particles by combining EDX and EBSD: application to workplace aerosols.

This paper discusses the combined use of electron backscatter diffraction (EBSD) and energy dispersive X-ray microanalysis (EDX) to identify unknown phases in particulate matter from different workplace aerosols. Particles of α-silicon carbide (α-SiC), manganese oxide (MnO) and α-quartz (α-SiO2) were used to test the method. Phase identification of spherical manganese oxide particles from ferromanganese production, with diameter less than 200 nm, was unambiguous, and phases of both MnO and Mn3O4 were identified in the same agglomerate. The same phases were identified by selected area electron diffraction (SAED) in transmission electron microscopy (TEM). The method was also used to identify the phases of different SiC fibres, and both ß-SiC and α-SiC fibres were found. Our results clearly demonstrate that EBSD combined with EDX can be successfully applied to the characterisation of workplace aerosols. Graphical abstract Secondary electron image of an agglomerate of manganese oxide particles collected at a ferromanganese smelter (a). EDX spectrum of the particle highlighted by an arrow (b). Indexed patterns after dynamic background subtraction from three particles shown with numbers in a

Electron microscopy of particles deposited in the lungs of nickel refinery workers.

The size, morphology, and chemical composition of particles deposited in the lungs of two nickel refinery workers were studied by scanning and transmission electron microscopy. The particles were extracted from the lung tissue by low-temperature ashing or by dissolution in tetramethylammonium hydroxide. The suitability of both sample preparation techniques was checked with reference materials. Both approaches lead to Fe-rich artifact particles. Low-temperature ashing leads to oxidation of small (diameter < 2 µm) metallic Ni and Ni sulfide particles, dissolution in tetramethylammonium hydroxide to removal of sulfate surface layers. Silicates and alumosilicates are the most abundant particle groups in the lungs of both subjects. From the various metal-dominated particle groups, Ni-rich particles are most abundant followed by Fe-rich and Ti-rich particles. Ni appears to be present predominantly as an oxide. Pure Ni metal and Ni sulfides were not observed. The presence of soluble Ni phases was not investigated as they will not be preserved during sample preparation. Based on their spherical morphology, it is estimated that a large fraction of Ni-rich particles (50-60 % by number) as well as Fe-rich and Cu-rich particles (27-45 %) originate from high-temperature processes (smelting, welding). This fraction is much lower for silicates (3-5 %), alumosilicates (1-2 %), and Ti-rich particles (9-11 %). The absence of metallic Ni particles most likely results from low exposure to this species. The absence of Ni sulfides may be either ascribed to low exposure or to fast clearance.

Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy.

Submicronic fungal fragments have been observed in in vitro aerosolization experiments. The occurrence of these particles has therefore been suggested to contribute to respiratory health problems observed in mold-contaminated indoor environments. However, the role of submicronic fragments in exacerbating adverse health effects has remained unclear due to limitations associated with detection methods. In the present study, we report the development of an indirect immunodetection assay that utilizes chicken polyclonal antibodies developed against spores from Aspergillus versicolor and high-resolution field emission scanning electron microscopy (FESEM). Immunolabeling was performed with A. versicolor fragments immobilized and fixed onto poly-l-lysine-coated polycarbonate filters. Ninety percent of submicronic fragments and 1- to 2-µm fragments, compared to 100% of >2-µm fragments generated from pure freeze-dried mycelial fragments of A. versicolor, were positively labeled. In proof-of-concept experiments, air samples collected from moldy indoor environments were evaluated using the immunolabeling technique. Our results indicated that 13% of the total collected particles were derived from fungi. This fraction comprises 79% of the fragments that were detected by immunolabeling and 21% of the spore particles that were morphologically identified. The methods reported in this study enable the enumeration of fungal particles, including submicronic fragments, in a complex heterogeneous environmental sample.

Involvement of IL-1 genes in the cellular responses to carbon nanotube exposure.

The interleukin-1 (IL-1) family has been implicated in cellular responses to nanoparticles including carbon nanotubes (CNTs). IL-1α and ß are key proinflammatory cytokines important in inflammatory and oxidative stress responses. The aim of this study was to characterize the role of IL-1 in cellular responses of CNTs in cells from IL-1α/ß wild type (IL1-WT) mice and cells with reduced inflammatory potential from IL-1α/ß deficient (IL1-KO) mice. Two multi-walled CNTs, CNT-1 containing long and thick fibers and CNT-2 containing short and thin fibers, were compared to UICC crocidolite asbestos fibers. Upon CNT exposure toxicity and apoptosis were affected differently in IL1-WT and IL1-KO cells. Upregulation of TNFα and IL-1α mRNA expression in IL1-WT cells was dependent on the type of CNT. On the contrary precursor IL-1α protein was downregulated after 24h. The mitogen-activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK) was activated in IL1-KO cells and regulated by CNTs, whereas no significant changes of extracellular regulated kinase (ERK) were observed when comparing IL1-WT and IL1-KO cells. In summary, the results presented here indicate that IL-1 contributes to the cellular and molecular effects of CNT exposure and that the type of CNT has an important effect on the cellular response.

Profile and Morphology of Fungal Aerosols Characterized by Field Emission Scanning Electron Microscopy (FESEM).

Fungal aerosols consist of spores and fragments with diverse array of morphologies; however, the size, shape, and origin of the constituents require further characterization. In this study, we characterize the profile of aerosols generated from Aspergillus fumigatus, A. versicolor, and Penicillium chrysogenum grown for 8 weeks on gypsum boards. Fungal particles were aerosolized at 12 and 20 L min-1 using the Fungal Spore Source Strength Tester (FSSST) and the Stami particle generator (SPG). Collected particles were analyzed with field emission scanning electron microscopy (FESEM). We observed spore particle fraction consisting of single spores and spore aggregates in four size categories, and a fragment fraction that contained submicronic fragments and three size categories of larger fragments. Single spores dominated the aerosols from A. fumigatus (median: 53%), while the submicronic fragment fraction was the highest in the aerosols collected from A. versicolor (median: 34%) and P. chrysogenum (median: 31%). Morphological characteristics showed near spherical particles that were only single spores, oblong particles that comprise some spore aggregates and fragments (<3.5 µm), and fiber-like particles that regroup chained spore aggregates and fragments (>3.5 µm). Further, the near spherical particles dominated the aerosols from A. fumigatus (median: 53%), while oblong particles were dominant in the aerosols from A. versicolor (68%) and P. chrysogenum (55%). Fiber-like particles represented 21% and 24% of the aerosols from A. versicolor and P. chrysogenum, respectively. This study shows that fungal particles of various size, shape, and origin are aerosolized, and supports the need to include a broader range of particle types in fungal exposure assessment.

Submicronic fungal bioaerosols: high-resolution microscopic characterization and quantification.

Submicronic particles released from fungal cultures have been suggested to be additional sources of personal exposure in mold-contaminated buildings. In vitro generation of these particles has been studied with particle counters, eventually supplemented by autofluorescence, that recognize fragments by size and discriminate biotic from abiotic particles. However, the fungal origin of submicronic particles remains unclear. In this study, submicronic fungal particles derived from Aspergillus fumigatus, A. versicolor, and Penicillium chrysogenum cultures grown on agar and gypsum board were aerosolized and enumerated using field emission scanning electron microscopy (FESEM). A novel bioaerosol generator and a fungal spores source strength tester were compared at 12 and 20 liters min(-1) airflow. The overall median numbers of aerosolized submicronic particles were 2 × 10(5) cm(-2), 2.6 × 10(3) cm(-2), and 0.9 × 10(3) cm(-2) for A. fumigatus, A. versicolor, and P. chrysogenum, respectively. A. fumigatus released significantly (P < 0.001) more particles than A. versicolor and P. chrysogenum. The ratios of submicronic fragments to larger particles, regardless of media type, were 1:3, 5:1, and 1:2 for A. fumigatus, A. versicolor, and P. chrysogenum, respectively. Spore fragments identified by the presence of rodlets amounted to 13%, 2%, and 0% of the submicronic particles released from A. fumigatus, A. versicolor, and P. chrysogenum, respectively. Submicronic particles with and without rodlets were also aerosolized from cultures grown on cellophane-covered media, indirectly confirming their fungal origin. Both hyphae and conidia could fragment into submicronic particles and aerosolize in vitro. These findings further highlight the potential contribution of fungal fragments to personal fungal exposure.

Characterisation of workplace aerosols in the manganese alloy production industry by electron microscopy.

Workplace aerosols in a combined FeMn and SiMn alloy smelter were studied by scanning and transmission electron microscopy. Special emphasis was placed on the characterisation of individual particles with diameters below 500 nm and on identification of the different manganese phases present in the workroom air. In high-carbon FeMn production, the submicron size fraction is dominated by MnO particles forming chain-like or compact agglomerates. Minor amounts of MnO(2), Mn(3)O(4), Mn(2)O(3) and Fe(3)O(4) are also observed. During production of SiMn, the submicron size fraction consists predominantly of MnSi particles, but small amounts of Mn(3)Si, Mn(6)Si and Mn(5)Si(2) are also found. Workplace aerosols from the manganese oxide refinement (MOR) process consist mostly of Mn oxides. Minor amounts of carbonaceous particles occurring as sheets, ribbons and as hollow carbon structures are observed along the whole production line. Carbonaceous particles are either amorphous or consist of poorly crystallised graphite. Particles with fibre morphology were encountered at all sampling locations but most prominently during tapping of FeMn with fibre concentrations between 0.1 and 0.7 per cm(3). The pronounced differences in particle composition along the production line clearly show that workers are exposed to a variety of Mn-containing species. MnO particles have a higher solubility than MnSi particles and are thus more bioaccessible, suggesting a higher risk of adverse health effects in the FeMn production than in the SiMn production.

Characterization of individual aerosol particles in workroom air of aluminium smelter potrooms.

Aerosol particles with aerodynamic diameters between 0.18 and 10 microm were collected in the workroom air of two aluminium smelter potrooms with different production processes (Soderberg and Prebake processes). Size, morphology and chemical composition of more than 2000 individual particles were determined by high resolution scanning electron microscopy and energy-dispersive X-ray microanalysis. Based on chemical composition and morphology, particles were classified into different groups. Particle groups with a relative abundance above 1%(by number) include aluminium oxides, cryolite, aluminium oxides-cryolite mixtures, soot, silicates and sea salt. In both production halls, mixtures of aluminium oxides and cryolite are the dominant particle group. Many particles have fluoride-containing surface coatings or show agglomerations of nanometer-sized fluoride-containing particles on their surface. The phase composition of approximately 100 particles was studied by transmission electron microscopy. According to selected area electron diffraction, sodium beta-alumina (NaAl(11)O(17)) is the dominant aluminium oxide and cryolite (Na(3)AlF(6)) the only sodium aluminium fluoride present. Implications of our findings for assessment of adverse health effects are discussed.

The heterogeneous composition of working place aerosols in a nickel refinery: a transmission and scanning electron microscope study.

Size, morphology and chemical composition of individual aerosol particles collected in a nickel refinery were analyzed by scanning electron microscopy and energy-dispersive X-ray microanalysis (EDX). The phase composition was determined by selected area electron diffraction and EDX in a transmission electron microscope. Most particles are heterogeneous on a nanometer scale and consist of various phases. Nickel phases observed in the roasting and anode casting departments include metallic nickel, bunsenite (NiO), trevorite (Ni,Cu)Fe2O4, heazlewoodite Ni3S2, godlevskite (Ni,Cu)9S8, orthorhombic NiSO4 and an amorphous Ni,Cu.Al,Pb sulfate of variable composition. Additional phases encountered include corundum (Al2O3), murdochite (PbCu6O8), hexagonal Na2SO4, anhydrite (CaSO4), graphite (C) and amorphous carbon. The implications of the occurrence of the different Ni phases and their nanometer size for the study of adverse health effects are explored.