Indirect immunodetection of fungal fragments by field emission scanning electron microscopy

Details

• Personal Author:
  Afanou KA ; Eduard W ; Green, Brett J. ; Hjeljord L ; Nayak, Ajay P. ; Skaar I ; Skogstad A ; Straumfors A ; Tronsmo A
• Description:
  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-microm fragments, compared to 100% of >2-microm 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. [Description provided by NIOSH]
• Subjects:
  Aerosols Air Pollutants, Occupational Air Pollution, Indoor Environmental Exposure Environmental Health Fungi Lung Lung Diseases Morbidity Occupational Health Particulate Matter Respiratory System Respiratory Tract Diseases Risk Assessment Risk Factors Safety

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• Keywords:
  Airborne-particles Environmental-exposure Exposure-levels Fungal-infections Indoor-air-pollution Indoor-environmental-quality Morbidity-rates Particulates Pulmonary-disorders Pulmonary-system Pulmonary-system-disorders Respiratory-infections Respiratory-system-disorders Risk-factors

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• ISSN:
  0099-2240
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  81
• Issue:
  17
• NIOSHTIC Number:
  nn:20046670
• Citation:
  Appl Environ Microbiol 2015 Sep; 81(17):5794-5803
• Contact Point Address:
  Eduard, Wijnand, National Institute of Occupational Health, Department of Chemical and Biological Work Environment, Oslo, Norway
• Email:
  wijnand.eduard@stami.no
• Federal Fiscal Year:
  2015
• NORA Priority Area:
  Services
• Peer Reviewed:
  True
• Source Full Name:
  Applied and Environmental Microbiology
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:b160c526d6fed3891e0aa5565f167afc3bafc7834194cf47f4e50ba10c31c20e4a77f87e00c30cc3be9d5bd68bc38b8b176aef06e5c6ecf51a97aecf11f28d3a
• Download URL:
  https://stacks.cdc.gov/view/cdc/202027/cdc_202027_DS1.pdf
• File Type:
  [PDF - 771.32 KB ]