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Applied and Environmental Microbiology, December 2005, p. 7948-7954, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7948-7954.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Determination of the Viability of Aeromonas hydrophila in Different Types of Water by Flow Cytometry, and Comparison with Classical Methods

Anna Pianetti,^1* Tania Falcioni,² Francesca Bruscolini,¹ Luigia Sabatini,¹ Elivio Sisti,⁴ and Stefano Papa^2,3

Toxicological, Hygienic and Environmental Science Institute, Urbino University, Urbino,¹ Center of Cytometry and Cytomorphology, Urbino University, Urbino,² Institute of Morphological Science, Urbino University, Urbino,³ Center for Environmental Study, Rimini, Italy⁴

Received 15 April 2005/ Accepted 25 August 2005

The presence of Aeromonas spp. in water can represent a risk for human health. Therefore, it is important to know the physiological status of these bacteria and their survival in the environment. We studied the behavior of a strain of Aeromonas hydrophila in river water, spring water, brackish water, mineral water, and chlorinated drinking water, which had different physical and chemical characteristics. The bacterial content was evaluated by spectrophotometric and plate count techniques. Flow cytometric determination of viability was carried out using a dual-staining technique that enabled us to distinguish viable bacteria from damaged and membrane-compromised bacteria. The traditional methods showed that the bacterial content was variable and dependent on the type of water. The results obtained from the plate count analysis correlated with the absorbance data. In contrast, the flow cytometric analysis results did not correlate with the results obtained by traditional methods; in fact, this technique showed that there were viable cells even when the optical density was low or no longer detectable and there was no plate count value. According to our results, flow cytometry is a suitable method for assessing the viability of bacteria in water samples. Furthermore, it permits fast detection of bacteria that are in a viable but nonculturable state, which are not detectable by conventional methods.

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* Corresponding author. Mailing address: Toxicological, Hygienic and Environmental Science Institute, Via S. Chiara, 27, University "Carlo BO," 61029 Urbino, Italy. Phone: 39 722 303544. Fax: 39 722 303541. E-mail: pianetti{at}uniurb.it.

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Applied and Environmental Microbiology, December 2005, p. 7948-7954, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7948-7954.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Falcioni, T., Papa, S., Gasol, J. M. (2008). Evaluating the Flow-Cytometric Nucleic Acid Double-Staining Protocol in Realistic Situations of Planktonic Bacterial Death. Appl. Environ. Microbiol. 74: 1767-1779 [Abstract] [Full Text]  
• Quiros, C., Herrero, M., Garcia, L. A., Diaz, M. (2007). Application of Flow Cytometry to Segregated Kinetic Modeling Based on the Physiological States of Microorganisms. Appl. Environ. Microbiol. 73: 3993-4000 [Abstract] [Full Text]