AAAR 33rd Annual Conference
October 20 - October 24, 2014
Rosen Shingle Creek
Orlando, Florida, USA
Abstract View
Analysis of Atmospheric Biological Particles with High-Resolution Microscopy Techniques
VERA SAMBUROVA, Alison Murray, Anna Gannet Hallar, Xufei Yang, Barbara Zielinska, Desert Research Institute
Abstract Number: 358 Working Group: Bioaerosols
Abstract Atmospheric biological particles (bacteria, fungal spores, plant pollen, etc.) and their fragments are known to comprise a significant portion of the ambient particulate matter. They play an important role in atmospheric processes and can cause adverse health effects, leading to chronic respiratory diseases. For example, pollen and plant debris may trigger allergic reactions in people, while bacterial endotoxins induce inflammatory responses. Biological particles are released in very large quantities into the atmosphere and may be transported over long distances. Therefore, there is an urgent need for bioaerosol characterization and quantitative analysis. In the present study atmospheric biological particles were collected at a high-elevation site and characterized with electron and epifluorescence microscopy techniques.
Airborne biological species were sampled at the Storm Peak Laboratory (SPL) on Mt. Werner, Steamboat, CO (3210 m AMSL). First, the shape and morphology of the collected bioaerosols were examined with a high-resolution electron microscopy (EM). Hitachi TM 1000 microscope was applied to take EM images directly from the collected filter punches. The rest of the aerosol filter sections were used to enumerate the biological particles. For this purpose, the samples were flushed with a sterile water to release particles from the filters and then the eluent was filtered on to 0.2 micron black polycarbonate filters that have low fluorescent background. The filters were stained with fluorescent dyes (e.g. 4-6- diamidino-2-phenylindole, DAPI and acridine orange, AO) to mark macromolecules (DNA or protein) in microbial, spore, and pollen grains. Following mounting on microscope slides, the filters were examined under the epifluorescence microscope (1000X magnification) and cell fractions were enumerated. Microbial abundance was back-calculated to cells per cubic meter of air. The microscopy results were compared with the data obtained from the culture-based analysis (rRNA genes sequencing technique).