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Applied and Environmental Microbiology, July 2005, p. 3524-3527, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3524-3527.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of a Simple and Rapid Fluorogenic Procedure for Identification of Vibrionaceae Family Members

Gary P. Richards,1* Michael A. Watson,1 and Salina Parveen2,{dagger}

United States Department of Agriculture, Agricultural Research Service, Microbial Food Safety Research Unit, Delaware State University, Dover, Delaware,1 Department of Agriculture and Natural Resources, Delaware State University, Dover, Delaware2

Received 3 November 2004/ Accepted 31 January 2005

We describe a simple colony overlay procedure for peptidases (COPP) for the rapid fluorogenic detection and quantification of Vibrionaceae from seawater, shellfish, sewage, and clinical samples. The assay detects phosphoglucose isomerase with a lysyl aminopeptidase activity that is produced by Vibrionaceae family members. Overnight cultures are overlaid for 10 min with membranes containing a synthetic substrate, and the membranes are examined for fluorescent foci under UV illumination. Fluorescent foci were produced by all the Vibrionaceae tested, including Vibrio spp., Aeromonas spp., and Plesiomonas spp. Fluorescence was not produced by non-Vibrionaceae pathogens. Vibrio cholerae strains O1, O139, O22, and O155 were strongly positive. Seawater and oysters were assayed, and 87 of 93 (93.5%) of the positive isolates were identified biochemically as Vibrionaceae, principally Vibrio vulnificus, Vibrio parahaemolyticus, Aeromonas hydrophila, Photobacterium damselae, and Shewanella putrefaciens. None of 50 nonfluorescent isolates were Vibrionaceae. No Vibrionaceae were detected in soil, and only A. hydrophila was detected in sewage. The COPP technique may be particularly valuable in environmental and food-testing laboratories and for monitoring water quality in the aquaculture industry.

* Corresponding author. Mailing address: USDA, ARS, Delaware State University, 1200 N. DuPont Hwy., James W. W. Baker Center, Dover, DE 19901. Phone: (302) 857-6419. Fax: (302) 857-6451. E-mail: grichard{at}desu.edu.

{dagger} Present address: University of Maryland Eastern Shore, 2112 Center for Food Science and Technology, Princess Anne, MD 21853.

Applied and Environmental Microbiology, July 2005, p. 3524-3527, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3524-3527.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Richards, G. P., Watson, M. A., Crane, E. J. III, Burt, I. G., Bushek, D. (2008). Shewanella and Photobacterium spp. in Oysters and Seawater from the Delaware Bay. Appl. Environ. Microbiol. 74: 3323-3327 [Abstract] [Full Text]  
  • RICHARDS, G. P., WATSON, M. A. (2006). A simple fluorogenic method to detect Vibrio cholerae and Aeromonas hydrophila in well water for areas impacted by catastrophic disasters.. Am J Trop Med Hyg 75: 516-521 [Abstract] [Full Text]  


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