Efficiency and specificity of CARD-FISH probes in detection of marine vibrios.

Abstract

Vibrio is a bacterial genus widely distributed in natural aquatic systems. Some Vibrio species can cause severe diseases in both marine organisms and humans. Previous studies revealed a link between the current climate change and increased incidence of the Vibrio-associated diseases recently causing sanitary, economic and/or ecological problems worldwide. The conventional culture-based methods (e.g. selection on TCBS agar) used to monitor the presence of Vibrio spp. in environmental samples are not always straightforward and can underestimate the number of cells, especially in microbial populations containing a fraction of 'dormant' cells (e.g. cells in the Viable but Non Culturable [VBNC] state). This problem can be overcome by using alternative culture-free approaches such as Catalysed Reporter Deposition-Fluorescence In situ Hybridization (CARD-FISH). To select an efficient CARD-FISH probe for detection of Vibrio spp. in environmental samples, we have assessed the most promising probes described in the literature by using both computer-assisted and experimental approaches. Our results demonstrate that the use of the optimized protocol along with a very specific probe, ViB572a, can offer the high sensitivity and selectivity of CARD-FISH detection of marine vibrios in natural seawater.