A New Field-Compatible Methodology for the Collection and Analysis of Fungal Fragments |
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Authors: | Sung-Chul Seo Sergey A. Grinshpun Yulia Iossifova Detlef Schmechel Carol Y. Rao Tiina Reponen |
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Affiliation: | 1. Center for Health-Related Aerosol Studies, Department of Environmental Health , University of Cincinnati , Cincinnati, Ohio, USA;2. National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention , Morgantown, West Virginia, USA |
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Abstract: | A field-compatible collection system was developed and tested for the collection and analysis of fungal fragments. The new collection system consists of two types of Sharp-Cut cyclone samplers (PM 2.5 and PM 1.0 ) and an after-filter. Fungal particles are collected into three size fractions: (1) spores ( > 2.5 μ m); (2) a fragment-spore mixture (1.0–2.5 μ m); and (3) submicrometer-sized fragments ( < 1.0 μ m). The system was laboratory-tested using polystyrene latex (PSL) particles and particulate matter aerosolized from sporulating Aspergillus versicolor and Stachybotrys chartarum cultures. In addition to the particle count measured with direct-reading instruments, the (1 → 3)- β -D-glucan content in each size fraction was determined with the Limulus Amebocyte Lysate (LAL) assay. Experiments conducted with PSL particles showed that the 50% cut-off values of the two cyclone samplers under the test conditions were 2.25 μ m and 1.05 μ m, respectively. No particle bounce onto the after-filter was observed when the total particle number entering the collection system was kept below 1.6 × 10 8 . The (1 → 3)- β -D-glucan assay of samples aerosolized from both fungal species suggested that surface area is an important factor for determining the (1 → 3)- β -D-glucan content in the entire size-range of particles. In conclusion, the new methodology is a promising tool for separating and analyzing fungal fragment samples. |
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