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Applied and Environmental Microbiology, March 2000, p. 1228-1232, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Flow Cytometry Method for Rapid Detection and Enumeration of Total Bacteria in Milk

Thusitha S. Gunasekera,* Paul V. Attfield, and Duncan A. Veal

Centre for Fluorimetric Applications in Biotechnology, Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia

Received 30 August 1999/Accepted 1 December 1999

Application of flow cytometry (FCM) to microbial analysis of milk is hampered by the presence of milk proteins and lipid particles. Here we report on the development of a rapid (<= 1-h) FCM assay based on enzymatic clearing of milk to determine total bacteria in milk. When bacteria were added to ultra-heat-treated milk, a good correlation (r >=  0.98) between the FCM assay and the more conventional methods of plating and direct microscopic counting was achieved. Raw milk data showed a significant correlation (P < 0.01) and a good agreement (r = 0.91) between FCM and standard plate count methods. The detection limit of the FCM assay was <= 104 bacteria ml of milk-1. This limit is below the level of detection required to satisfy legislation in many countries and states.

* Corresponding author. Mailing address: Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia. Phone: 61-2-98508157. Fax: 61-2-98508253. E-mail: tgunasek{at}rna.bio.mq.edu.au.

Applied and Environmental Microbiology, March 2000, p. 1228-1232, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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