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Nucleic Acid Tests for Toxigenic Phytoplankton Species in Irish Waters A collaborative research project which commenced in July, 2005 involving researchers at the National Diagnostics Centre, NUI, Galway (NDC) and the Marine Environment and Food Safety Services of the Irish Marine Institute (MI) is focusing on developing nucleic acid diagnostic tests (NAD) for the detection and identification of toxigenic phytoplankton species of concern in particular, Dinophysis and Pseudo-nitzchia sps. in Irish waters. Annually, the MI analyse more than 2000 water samples from shellfish production sites and salmon farms for the presence of toxic algal species as part of their monitoring programme. Current identification of species relies on microscopy which is very useful in providing a global view of phytoplankton species present in water samples but cannot precisely identity some important toxic species. Molecular methods can be applied to identify toxic species and to discriminate between closely related species. Internationally, phytoplankton monitoring programmes are successfully applying molecular methods to support their services. As part of a previous collaborative project funded under the Programme for Research and Training in Third Level Institutions PRTLI (2002-2005) researchers at the Martin Ryan Institute at NUI, Galway and the NDC developed NAD tests including FISH DNA probe tests and real-time PCR assays for identification of Alexandrium species, A. tamarense and A. minutum and demonstrated their application to species identification in wild samples. In the current project, two NAD formats are being developed, a low-cost low-tech direct DNA probe based test which can be applied directly to a sample to identify toxic species. Real-time PCR based tests capable of rapid, sensitive and high-throughput detection of toxic species will also be developed. In this phase of the project samples are being collected from around the Irish coast, toxic species isolated and cultured where possible. Selected genomic regions including the small ribosomal (SSU), large ribosomal subunit (LSU) and the internally transcribed spacer regions (ITS) are being sequenced to provide sequence information for design of NADs. DNA probe design and real-time PCR assay development work has commenced. For further information contact: Majella.Maher@nuigalway.ie or joe.silke@marine.ie
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