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Annals of Occupational Hygiene Advance Access originally published online on August 12, 2009
Annals of Occupational Hygiene 2009 53(8):859-868; doi:10.1093/annhyg/mep060
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© The Author 2009. Published by Oxford University Press on behalf of the British Occupational Hygiene Society

Direct Detection of Salmonella Cells in the Air of Livestock Stables by Real-Time PCR

Kerstin Fallschissel1, Peter Kämpfer1 and Udo Jäckel2,*

1 Institute for Applied Microbiology, Justus-Liebig University of Giessen, Heinrich-Buff-Ring 26-32, 34637 Giessen, Germany
2 Federal Institute for Occupational Safety and Health, Nöldnerstraße 40-42; 10317 Berlin, Germany

* Author to whom correspondence should be addressed. Tel: +49 3051548-4788; fax: +49 3051548-4171; e-mail: jaeckel.udo{at}baua.bund.de

A SYBR® Green real-time quantitative polymerase chain reaction (qPCR) assay for specific detection and quantification of airborne Salmonella cells in livestock housings is presented. A set of specific primers was tested and validated for specific detection and quantification of Salmonella-specific invA genes of DNA extracted from bioaerosol samples. Application of the method to poultry house bioaerosol samples showed concentrations ranging from 2.2 x 101 to 3 x 106 Salmonella targets m–3 of air. Salmonella were also detected by a cultivation-based approach in some samples, but concentrations were two to three magnitudes lower than the concentrations detected by molecular biological results. Specificity of results was demonstrated by cloning analyses of PCR products, which were exclusively assigned to the genus Salmonella. However, by molecular methods, microorganisms are detected independently of their viability status, leading to an overestimation of concentration. Hence, the survival rate of Salmonella cells was measured on filter surfaces during filtration samplings where 82% of the cells died within 20 min of filtration. The results clearly show the specificity and practicability of the established qPCR assay for analysis and quantification of salmonellae in bioaerosols. The results demonstrate airborne Salmonella workplace concentrations in poultry production of up to 3.3% of 4',6-Diamidino-2-phenylindole-counted total cell numbers.

Keywords: bioaerosol • DNA extraction efficiency • livestock stables • poultry • real-time PCR • Salmonella • sampling efficiency • SYBR Green

Received March 16, 2009; in final form July 16, 2009


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