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Monitoring toxic microalgae Ostreopsis (dinoflagellate) species in coastal waters of the Mediterranean Sea using molecular PCR-based assay combined with light microscopy
Authors:Cecilia Battocchi  Magda Vila  Samuela Capellacci  Albert Reñé  Antonella Penna
Institution:a Dipartimento di Scienze Biomolecolari, Sez. Biol. Amb., Università di Urbino, Viale Trieste 296, 61100 Pesaro, Italy
b Dipartimento di Scienze del Mare, Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy
c Institut de Ciències del Mar ICM, CSIC, Passeig Marítim de la Barceloneta 37-43, 08003 Barcelona, Spain
d Dipartimento di Biologia, Università di Roma “Tor Vergata”, Via della Ricerca Scientifica, 00133 Rome, Italy
Abstract:A molecular PCR-based assay was developed and applied to macrophyte and seawater samples containing mixed microphytobenthic and phytoplanktonic assemblages, respectively, in order to detect toxic Ostreopsis species in Mediterranean Sea. The specificity and sensitivity of the molecular PCR assay were assessed with both plasmidic and genomic DNA of the target genus or species using taxon-specific primers in the presence of background macrophyte DNA. The PCR molecular technique allowed rapid detection of the Ostreopsis cells, even at abundances undetectable within the resolution limit of the microscopy technique. Species-specific identification of Ostreopsis was determined only by PCR-based assay, due to the inherent difficulty of morphological identification in field samples. In the monitoring of the toxic Ostreopsis blooms PCR-based methods proved to be effective tools complementary to microscopy for rapid and specific detection of Ostreopsis and other toxic dinoflagellates in marine coastal environments.
Keywords:Coastal areas  HAB species  Mediterranean Sea  Monitoring  Ostreopsis  PCR
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