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Applied and Environmental Microbiology, June 2003, p. 3510-3516, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3510-3516.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of Growth Indices from Radiometric Culture for Quantification of Sheep Strains of Mycobacterium avium subsp. paratuberculosis

Leslie A. Reddacliff,1,{dagger}* Paul J. Nicholls,2 Aparna Vadali,2,{ddagger} and Richard J. Whittington2,{ddagger}

Faculty of Veterinary Science, University of Sydney, Sydney,1 Elizabeth Macarthur Agricultural Institute, New South Wales Agriculture, Camden, New South Wales 2570, Australia2

Received 2 December 2002/ Accepted 12 March 2003

A simple method for using growth indices from radiometric BACTEC cultures was evaluated for the enumeration of Australian sheep strains of Mycobacterium avium subsp. paratuberculosis. The numbers of viable organisms in inocula were determined by end-point titration in BACTEC cultures. Growth indices were measured by using a BACTEC 460 machine. There was a linear relationship between the number of days taken for the cumulative growth index to reach 1,000 (dCGI1000) and log10 inoculum size. The use of dCGI1000 was shown to be as effective as the use of growth index data from the entire growth cycle for the estimation of inoculum size. For particular isolates characterized by end-point titration, the dCGI1000 of a single BACTEC vial provided estimates of viable numbers within narrow prediction limits. Predictive relationships were also established for the enumeration of M. avium subsp. paratuberculosis from field samples by using the dCGI1000 of a single BACTEC vial, with prediction limits of ±1 to 2 log units. Organisms from feces or contaminated soil grew more slowly than those from cultures or tissues, and separate equations were developed for enumeration from these sources.

* Corresponding author. Present address: Elizabeth Macarthur Agricultural Institute, New South Wales Agriculture, PMB 8, Camden, New South Wales 2570, Australia. Phone: (61) 0246406314. Fax: (61) 0246406384. E-mail: leslie.reddacliff{at}agric.nsw.gov.au.

{dagger} Present address: Diadexus Inc., South San Francisco, CA 94080.

{ddagger} Present address: Faculty of Veterinary Science, University of Sydney, Camden, New South Wales 2570, Australia.

Applied and Environmental Microbiology, June 2003, p. 3510-3516, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3510-3516.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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