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Algal toxins in Scottish waters: the first association of domoic acid with the diatom Pseudo-nitzschia pseudodelicatissima complex in the North East of Scotland Eileen Bresnan, Jean-Pierre Lacaze, Sheila Fraser, Lyndsay Brown, Ana-Luisa Amorim and Kathryn Cook Marine Scotland Science, Marine Laboratory, 375 Victoria Road, Aberdeen AB11 9DB E-mail: [email protected] Introduction How to identify Pseudo-nitzschia? • Diatom genus Pseudo-nitzschia (Peragallo) • Light microscopy (LM) cannot identify Pseudonitzschia cells to species level • Routinely observed in Scottish waters • • Two species within genus (P. australis and P. seriata) confirmed as producers of domoic acid (DA)1 LM can only separate cells into two different size classes (</> 5µm diameter) • Electron microscopy or molecular methods required to confirm identification • DA toxin poisoning • High concentrations of DA closed offshore scallop fisheries from 1998 – 20051 • DA detected in faeces and urine of seals along the east coast of Scotland 2 • 2008 – Study at Stonehaven ecosystem monitoring site considered if zooplankton could act as a vector for DA in Scottish waters responsible for amnesic shellfish Light micrograph of Pseudo-nitzschia cells Enlarged central portions of Pseudo-nitzschia cells P. seriata Transmission electron micrograph of Pseudo-nitzschia cells P. australis Algal toxins in Scottish waters: the first association of domoic acid with the diatom Pseudo-nitzschia pseudodelicatissima complex in the North East of Scotland Study site • Stonehaven monitoring site sampled weekly since 1997 • Temperature, salinity, and plankton measured • Calanus was sampled during 2008 for DA analysis • Liquid chromatography – mass spectrometry – mass spectrometry (LC-MS-MS) was used Toxin data is nutrients Latitude 60N 58N Stonehaven The copepod Calanus Pseudo-nitzschia cells counts and DA concentrations in Calanus at the Stonehaven monitoring site in 2008. 56N 54N -8W -4W Longitude • DA does accumulate in copepods at Stonehaven • A maximum concentration of 11 ρg DA. copepod-1 was recorded in July • Diversity of the Pseudo-nitzschia bloom during July was examined 0W Algal toxins in Scottish waters: the first association of domoic acid with the diatom Pseudo-nitzschia pseudodelicatissima complex in the North East of Scotland Structure of Pseudo-nitzschia cells from July bloom Identification of Pseudo-nitzschia cells Whole Pseudo-nitzschia cell • A JEOL JEM 1400 Plus TEM was used to examine cells • Morphological structures on Pseudo-nitzschia were measured • Cells were identified as the recently described plurisecta3 • This is a species within the P. pseudodelicatissima species complex • Blooms of this species were previously observed in Scottish waters following an extensive Karenia mikimotoi bloom. • This is the first association of this species with DA accumulation in Scottish waters Pseudo-nitzschia Acknowledgements Central Interspace (CI) Poroids Fibulae • The authors would like to thank Mr. Kevin McKenzie at the Electron Microscopy Unit, Institute of Medical Sciences, University of Aberdeen for his assistance. • This project was funded as part of the Scottish Government’s schedule of service ST03p and ROAME AE1198. Striae Morphological measurements Length (µm) Width (µm) CI 68 1.2 – 1.5 Y Fibulae (in 10µm) Striae (in 10µm) Poroids (in 1µm) 23-25 39 – 43 5-6 References • • • J., Green D., Davidson K., Bolch C. and Bates S. (2004) J. Phycol., 40, 622 – 630. A. and Frame E. (2010), Harmful Algae, 9, 489 – 493. 3Orive E., David H. and Miguel I. (2013), J. Phycol. 49, 1192 - 1206 1Fehling 2Hall