Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 11/21/2004
Publication Date: 1/17/2005
Citation: Richards, G.P., Watson, M.A., Burt, I.G., Bushek, D. 2005. Detection of vibrios in oysters crassostrtea virginica and seawater using a new, rapid, simple, and quantitative enzyme-based assay. (Abstract). Aquaculture America. Paper # 8128.
Technical Abstract: The Vibrionaceae family of bacteria contains important human and fish pathogens. We developed a rapid, simple, and quantitative detection method for members of the Vibrionaceae family. The assay, referred to as the colony overlay procedure for peptidases (COPP), is based on the rapid and inexpensive detection of an enzyme found in Vibrionaceae family members. Membranes are overlayed onto overnight cultures of bacteria and after 10 min, the membranes are exposed to long-wavelength UV (e.g., a black light) and examined for fluorescent spots that correspond to the overlayed colonies. All members of the Vibrionaceae family tested to date (eleven species), including bacteria from the genera Vibrio (V. vulnificus, V. parahaemolyticus, V. cholerae, V. alginolyticus, V. harveyi, V. fluvialis, and others), Photobacterium, Aeromonas, and Shewanella, produced fluorescent foci. Nine non-vibrios, including E. coli, Salmonella and Shigella species, Klebsiella, Enterobacter, Staphylococcus and Streptococcus did not produce fluorescence. A survey of Eastern oysters, Crassostrea virginica, and seawater from oyster harvesting sites in New Jersey demonstrated the effectiveness of this new technique. Since the Vibrionaceae are often associated with diseases to aquaculture species, the COPP assay would facilitate the monitoring of seawater, fish and shellfish for total Vibrio levels. The use of this method to determine total Vibrio counts in seawater or in aquaculture species could be used to detect increases in bacterial levels and forewarn the producer or processor of potential problems so that remedial actions may be initiated. Since the test does not require sophisticated or expensive equipment, the COPP method may be a practical solution for monitoring vibrios in aquaculture settings.