FUENTES, S*; RICK, H; NOEL, J; University of Louisiana at Lafayette: Development of Real-Time PCR assays for rapid detection of Cylindrospermopsis raciborskii

Blooms caused by toxic blue-green algae have repeatedly produced episodes of wild and domestic animal illness and death. The responsible cyanobacteria belong to about 40 genera including the most recently found toxin-producing cyanobacterium, Cylindrospermopsis. Since 2002, Cylindrospermopsis raciborskii has been abundant in water samples of Caernarvon/Breton Sound (LA), posing a potential risk of a bloom of this species in that area. Recently (10/04) we found abundant biomasses of this species in the whole Atchafalaya Basin, from Krotz Springs to Morgan City. Monitoring these areas for the presence of Cylindrospermopsis is essential in assessing the potential for bloom formation. However, detection and quantification of Cylindrospermopsis is problematic since it is not easily distinguished from suspended sediment or other types of algae. Given this, we are developing a real-time PCR assay for detection of Cylindrospermopsis raciborskii. In this assay, detection of amplified target DNA requires annealing of fluorescent-labeled probes, resulting in added level of specificity compared with assays based on traditional PCR methodology. Using this sensitive technique it will be possible to detect even low densities of this species instantly, allowing the creation of real-time species abundance distribution maps, enabling managers to respond immediately to increases in biomass of this harmful species and for timely notification of possible health risks to the public