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Applied and Environmental Microbiology, April 2005, p. 1946-1952, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.1946-1952.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Detection of Mycobacterium bovis and Mycobacterium bovis BCG (Pasteur) in Soil

Jamie S. Young,1 Eamonn Gormley,2 and Elizabeth M. H. Wellington1*

Department of Biological Sciences, University of Warwick, Coventry, United Kingdom,1 Department of Large Animal Clinical Studies, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin, Ireland2

Received 17 February 2004/ Accepted 2 November 2004

PCR primers specific for the Mycobacterium tuberculosis complex were used to detect the presence of Mycobacterium bovis BCG (Pasteur) in soil microcosms and Mycobacterium bovis in environmental samples taken from a farm in Ireland with a history of bovine tuberculosis. M. bovis genes were detected in soil at 4 and 21 months after possible contamination. Gene levels were found in the range of 1 x 103 to 3.6 x 103 gene copies g of soil–1, depending on the sampling area. Areas around badger setts had the highest levels of detectable genes and were shown to have the highest levels of gene persistence. M. bovis-specific 16S rRNA sequences were detected, providing evidence of the presence of viable cells in Irish soils. Studies of DNA turnover in soil microcosms proved that dead cells of M. bovis BCG did not persist beyond 10 days. Further microcosm experiments revealed that M. bovis BCG survival was optimal at 37°C with moist soil (–20 kPa; 30% [vol/wt]). This study provides clear evidence that M. bovis can persist in the farm environment outside of its hosts and that climatic factors influence survival rates.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, United Kingdom. Phone: 44 247653184. Fax: 44 2476523701. E-mail: e.m.h.wellington{at}warwick.ac.uk.

Applied and Environmental Microbiology, April 2005, p. 1946-1952, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.1946-1952.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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