Microbial contaminants including nontuberculous mycobacteria (NTM) in metalworking fluids (MWFS) have been implicated in occupational respiratory illnesses. Our broader goals are to develop and apply practical DNA-based approaches for real-time detection, quantitation, and identification of mycobacteria in MWF for understanding the prevalent strains and their survivability in commercial MWFS.
The specific aims are (i) to develop polymerase chain reaction (PCR)-based protocols for real-time detection and quantitation of NTM in water-based MWF; (ii). to PCR screen field samples of different commercial formulations of water-based MWFs for NTM followed by strain-specific identification of the NTM isolates using molecular typing methods; (iii) to investigate viability of the identified NTM strains in MWF using quantitative PCR. A Mycobacterium genus-specific PCR-based method will be optimized for real-time detection of NTM in water-based MWF. The method will be used as a basis for developing a quantitative PCR protocol for quantitation of NTM in field MWF samples. NTM strains will be isolated from MWF samples in use across the industries and in selected plants linked with hypersensitivity pneumonitis cases. The isolates will be identified at the species level by 16S ribosomal gene amplicon sequencing and at the strain level based on genome fingerprinting using pulsed field gel electrophoresis (PFGE) or rapid amplification of polymorphic DNA-PCR (RAPD-PCR) technique. Effect of strain type and biocide load on viability of the selected strains in MWF will be investigated in simulated as well as field conditions using quantitative PCR. The resulting information will help NORA's objectives by providing practical methods for microbial exposure assessment and database for developing intervention strategies related to MWF exposures.

Agency
National Institute of Health (NIH)
Institute
National Institute for Occupational Safety and Health (NIOSH)
Type
Research Project (R01)
Project #
5R01OH007364-02
Application #
6598106
Study Section
Safety and Occupational Health Study Section (SOH)
Program Officer
Karr, Joan
Project Start
2001-09-30
Project End
2004-09-29
Budget Start
2002-09-30
Budget End
2003-09-29
Support Year
2
Fiscal Year
2002
Total Cost
$223,670
Indirect Cost
Name
University of Cincinnati
Department
Public Health & Prev Medicine
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Johansson, Elisabet; Boivin, Gregory P; Yadav, Jagjit S (2017) Early immunopathological events in acute model of mycobacterial hypersensitivity pneumonitis in mice. J Immunotoxicol 14:77-88
Chandra, Harish; Yadav, Jagjit S (2016) Human leukocyte antigen (HLA)-binding epitopes dataset for the newly identified T-cell antigens of Mycobacterium immunogenum. Data Brief 8:1069-71
Chandra, Harish; Yadav, Jagjit S (2016) T-cell antigens of Mycobacterium immunogenum, an etiological agent of occupational hypersensitivity pneumonitis. Mol Immunol 75:168-77
Chandra, Harish; Lockey, James; Yadav, Jagjit S (2015) Novel antigens of Mycobacterium immunogenum relevant in serodiagnosis of occupational hypersensitivity pneumonitis in machinists. Ann Allergy Asthma Immunol 114:525-6
Kapoor, Renuka; Selvaraju, Suresh B; Yadav, Jagjit S (2014) Extended tracking of the microbial community structure and dynamics in an industrial synthetic metalworking fluid system. FEMS Microbiol Ecol 87:664-77
Chandra, Harish; Yadav, Ekta; Yadav, Jagjit S (2013) Alveolar macrophage innate response to Mycobacterium immunogenum, the etiological agent of hypersensitivity pneumonitis: role of JNK and p38 MAPK pathways. PLoS One 8:e83172
Selvaraju, Suresh B; Khan, Izhar U H; Yadav, Jagjit S (2011) Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to formaldehyde and non-formaldehyde biocides in semi-synthetic metalworking fluids. Int J Mol Sci 12:725-41
Kapoor, Renuka; Yadav, Jagjit S (2010) Development of a rapid ATP bioluminescence assay for biocidal susceptibility testing of rapidly growing mycobacteria. J Clin Microbiol 48:3725-8
Gupta, Manish K; Subramanian, Venkataraman; Yadav, Jaglit S (2009) Immunoproteomic identification of secretory and subcellular protein antigens and functional evaluation of the secretome fraction of Mycobacterium immunogenum, a newly recognized species of the Mycobacterium chelonae-Mycobacterium abscessus group. J Proteome Res 8:2319-30
Selvaraju, Suresh B; Kapoor, Renuka; Yadav, Jagjit S (2008) Peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) assay for specific detection of Mycobacterium immunogenum and DNA-FISH assay for analysis of pseudomonads in metalworking fluids and sputum. Mol Cell Probes 22:273-80

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