Specific Detection and Quantification of Culturable and Non-Culturable Mycobacteria in Metalworking Fluids by Fluorescence-Based Methods

Details

• Personal Author:
  Khan IU ; Selvaraju SB ; Yadav JS
• Description:
  AIMS: To optimize and evaluate fluorescence microscopy assays for specific assessment of mycobacteria and co-contaminants, including culturable and non-culturable sub-populations, in metalworking fluids (MWF). Methods and RESULTS: Auramine-O-rhodamine (AR) staining and LIVE/DEAD BacLight Bacterial Viability staining (L/D staining) were adapted and evaluated for detection/quantification and differentiation (viable vs non-viable) of the MWF-associated mycobacteria and the background bacterial flora, respectively. The AR staining method was found to be specific to MWF mycobacteria with a minimum detection limit of 10 cells ml(-1) and was comparable to the QPCR in quantification efficiency in MWF matrix. The L/D staining-based microscopy allowed differential quantification of viable vs non-viable cells. In general, a 3-log difference was observed between the L/D microscopy count and culture count accounting for the presence of non-culturable fraction in the bacterial population in in-use MWF. The optimized AR staining- and the L/D staining-based microscopy methods have the potential for rapid, specific and differential assessment (viable vs non-viable) of MWF-associated mycobacteria and co-contaminants in field MWF. SIGNIFICANCE AND IMPACT OF THE STUDY: Early detection of MWF mycobacteria by rapid, low-cost, less-skill intensive and culture-independent fluorescence-based microscopy methods will facilitate timely intervention to protect the machine workers from occupational hazards. [Description provided by NIOSH]
• Subjects:
  Hazardous Substances Manufacturing And Industrial Facilities Metals Metal Workers Microbiology Occupational Diseases Occupational Exposure Occupational Health Safety Workplace
• Keywords:
  Author Keywords: Epifluorescence Microscopy Bacterial-cultures Live/Dead Baclight Staining Machine-operators Metalworking Metalworking-fluids Metalworking-industry Metalworking Fluid Microscopic-analysis Mycobacterium Immunogenum Non-culturable Occupational-diseases Occupational-exposure Occupational-health Quantitative-analysis Real-time PCR Toxic-materials Viable But Non-culturable Worker-health Workplace-monitoring

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• ISSN:
  0266-8254
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  Missouri ; Ohio ; OSHA Region 5 ; OSHA Region 7
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  451-456
• Volume:
  47
• Issue:
  5
• NIOSHTIC Number:
  nn:20035055
• Citation:
  Lett Appl Microbiol 2008 Nov; 47(5):451-456
• Contact Point Address:
  Jagjit S. Yadav, Environmental Genetics and Molecular Toxicology Division, Department of Environmental Health, College of Medicine, University of Cincinnati, Cincinnati, OH 45267-0056
• Email:
  jagjit.yadav@uc.edu
• Federal Fiscal Year:
  2009
• Performing Organization:
  University of Cincinnati
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  Letters in Applied Microbiology
• End Date:
  20170731
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:e924cf5e9bdd36e3eacb31090a02e85e54e11dff6b6efe4cb6f1c41f566a71901c2ee7025da12d0e9f14eeebca3077649e31d50e3fc33606e7cf5b4a1ade62a6
• Download URL:
  https://stacks.cdc.gov/view/cdc/187038/cdc_187038_DS1.pdf
• File Type:
  [PDF - 264.20 KB ]