Harmful algal blooms (HABs) occur worldwide, causing health problems and economic damages to fisheries and tourism. Monitoring agencies are therefore essential, yet monitoring is based only on time-consuming light microscopy, a level at which a correct identification can be limited by insufficient morphological characters. The project MIDTAL (Microarray Detection of Toxic Algae)—an FP7-funded EU project—used rRNA genes (SSU and LSU) as a target on microarrays to identify toxic species. Furthermore, toxins were detected with a newly developed multiplex optical Surface Plasmon Resonance biosensor (Multi SPR) and compared with an enzyme-linked immunosorbent assay (ELISA). In this study, we demonstrate the latest generation of MIDTAL microarrays (version 3) and show the correlation between cell counts, detected toxin and microarray signals from field samples taken in Arcachon Bay in France in 2011. The MIDTAL microarray always detected more potentially toxic species than those detected by microscopic counts. The toxin detection was even more sensitive than both methods. Because of the universal nature of both toxin and species microarrays, they can be used to detect invasive species. Nevertheless, the MIDTAL microarray is not completely universal: first, because not all toxic species are on the chip, and second, because invasive species, such as Ostreopsis, already influence European coasts.
References
[1]
Hallegraeff, G.M. Harmful algal blooms: A global overview. In Manual on Harmful Marine Microalgae; IOC-UNESCO: Paris, France, 2003; Volume 11, pp. 25–49.
[2]
Moestrup, ？.; Akselman, R.; Cronberg, G.; Elbraechter, M.; Fraga, S.; Halim, Y.; Hansen, G.; Hoppenrath, M.; Larsen, J.; Lundholm, N.; Nguyen, L.N.; Zingone, A. IOC-UNESCO Taxonomic Reference List of Harmful Micro Algae. Available online: http://www.marinespecies.org/HAB (accessed on 24 December 2012).
[3]
Yasumoto, T.; Murata, M.; Oshima, Y.; Sano, M.; Matsumoto, G.K.; Clardy, J. Diarrhetic shellfish toxins. Tetrahedron 1985, 41, 1019–1025, doi:10.1016/S0040-4020(01)96469-5.
Gómez, F. Phytoplankton invasions: Comments on the validity of categorizing the non-indigenous dinoflagellates and diatoms in European seas. Mar. Pollut. Bull. 2008, 56, 620–628, doi:10.1016/j.marpolbul.2007.12.014.
[6]
DeSantis, T.Z.; Stone, C.E.; Murray, S.R.; Moberg, J.P.; Andersen, G.L. Rapid quantification and taxonomic classification of environmental DNA from both prokaryotic and eukaryotic origins using a microarray. FEMS Microbiol. Lett. 2005, 245, 271–278, doi:10.1016/j.femsle.2005.03.016.
Yergeau, E.; Arbour, M.; Brousseau, R.; Juck, D.; Lawrence, J.R.; Masson, L.; Whyte, L.G.; Greer, C.W. Microarray and real-time PCR analyses of the responses of high-arctic soil bacteria to hydrocarbon pollution and bioremediation treatments. Appl. Environ. Microbiol. 2009, 75, 6258–6267.
[9]
Gescher, C.; Metfies, K.; Medlin, L.K. The ALEX CHIP—Development of a DNA chip for identification and monitoring of Alexandrium. Harmful Algae 2008, 7, 485–494, doi:10.1016/j.hal.2007.11.001.
[10]
Brodie, E.L.; DeSantis, T.Z.; Joyner, D.C.; Baek, S.M.; Larsen, J.T.; Andersen, G.L.; Hazen, T.C.; Richardson, P.M.; Herman, D.J.; Tokunaga, T.K.; Wan, J.M.; Firestone, M.K. Application of a high-density oligonucleotide microarray approach to study bacterial population dynamics during uranium reduction and reoxidation. Appl. Environ. Microbiol. 2006, 72, 6288–6298.
[11]
Brodie, E.L.; DeSantis, T.Z.; Parker, J.P. M.; Zubietta, I.X.; Piceno, Y.M.; Andersen, G.L. Urban aerosols harbor diverse and dynamic bacterial populations. PNAS 2007, 104, 299–304.
[12]
Delehanty, J.B.; Ligler, F.S. A microarray immunoassay for simultaneous detection of proteins and bacteria. Anal. Chem. 2002, 74, 5681–5687, doi:10.1021/ac025631l.
[13]
Lee, D.-Y.; Seto, P.; Korczak, R. DNA microarray-based detection and identification of waterborne protozoan pathogens. J. Microbiol. Meth. 2010, 80, 129–133, doi:10.1016/j.mimet.2009.11.015.
[14]
Lewis, J.; Medlin, L.K.; Raine, R. MIDTAL (Microarrays for the Detection of Toxic Algae): A Protocol for a Successful Microarray Hybridisation and Analysis, 1st ed.; Gantner Verlag: Liechtenstein, Germany, 2012.
[15]
Medlin, L.K.; Schmidt, K. Molecular probes improve the taxonomic resolution of cryptophyte abundance in Arcachon Bay. Vie et Milieu 2010, 60, 9–15.
[16]
Kegel, J.U.; Del Amo, Y.; Medlin, L.K. Introduction to project MIDTAL: Its methods and samples from Arcachon Bay, France. Environ. Sci. Pollut. Res. 2012, doi:10.1007/s11356-012-1299-9.
[17]
Campbell, K.; McGrath, T.; Sj？lander, S.; Hanson, T.; Tidare, M.; Jansson, O.; Moberg, A.; Mooney, M.; Elliott, C.; Buijs, J. Use of a novel micro-fluidic device to create arrays for multiplex analysis of large and small molecular weight compounds by surface plasmon resonance. Biosens. Bioelectron. 2011, 26, 3029–3036, doi:10.1016/j.bios.2010.12.007.
Metfies, K.; Borsutzki, P.; Gescher, C.; Medlin, L.K.; Frickenhaus, S. Phylochipanalyser—A program for analysing hierarchical probe sets. Mol. Ecol. Resour. 2008, 8, 99–102, doi:10.1111/j.1471-8286.2007.01927.x.
[20]
Dittami, S.M.; Edvardsen, B. GPR-Analyzer: A simple tool for quantitative analysis of hierarchical multispecies microarrays. Environ. Sci. Pollut. Res. 2012, doi:10.1007/s11356-012-1051-5.
[21]
Hasle, G.R. Nitzschia and Fragilariopsis Species Studied in the Light and Electron Microscopes. II. The Group Pseudonitzschia; Universitetsforlaget: Oslo, Norway, 1965.
[22]
Hasle, G.R.; Syvertsen, E.E. Marine diatoms. In Identifying Marine Phytoplankton; Academic Press: San Diego, CA, USA, 1997; pp. 5–385.
[23]
McNamee, S.; Elliott, C.; Delahaut, P.; Campbell, K. Multiplex biotoxin surface plasmon resonance method for marine biotoxins in algal and seawater samples. Environ. Sci. Pollut. Res. 2012, doi:10.1007/s11356-012-1329-7.
[24]
Taylor, J.; Kegel, J.U.; Lewis, J.; Medlin, L.K. Validation of the detection of Alexandrium spp using specific RNA probes tested in a microarray format: Calibration of signal based on variability of RNA content with environmental conditions. Harmful Algae 2013. submitted.
[25]
Blanco, E.P.; Hagstr？m, J.; Salomon, P.S.; Graneli, E. Detection of Heterosigma akashiwo (Hada) using specific RNA probes: Variability of RNA content with environmental conditions. Harmful Algae 2013. submitted.
[26]
Dittami, S.M.; Hostyeva, V.; Egge, E.S.; Kegel, J.U.; Eikrem, W.; Edvardsen, B. Seasonal dynamics of harmful algae in outer Oslofjorden monitored by microarray, qPCR, and microscopy. Environ. Sci. Pollut. Res. 2013, doi:10.1007/s11356-012-1392-0.
[27]
Naustvoll, L.-J. NOBANIS—Invasive Alien Species Fact Sheet Pseudochattonella farcimen—From: Online Database of the North Eurpean and Baltic Network on Invasive Alien Species—NOBANIS. Available online: http://www.nobanis.org (accessed on 24 December 2012).
[28]
Glé, C.; Del Amo, Y.; Sautour, B.; Laborde, P.; Chardy, P. Variability of nutrients and phytoplankton primary production in a shallow macrotidal coastal ecosystem (Arcachon Bay, France). Estuar. Coast. Shelf Sci. 2008, 76, 642–656, doi:10.1016/j.ecss.2007.07.043.
[29]
McCoy, G.R.; Touzet, N.; Fleming, G.T.; Raine, R. An evaluation of the applicability of microarrays for monitoring toxic algae in Irish coastal waters. Environ. Sci. Pollut. Res. 2012, doi:10.1007/s11356-012-1294-1.
[30]
Swan River Trust Karlodinium Micrum Algal Bloom, March 2005. Fact Sheet. Available online: http://www.wa.canoe.org.au/?Page=7808 (accessed on 24 December 2012).
[31]
Tillmann, U.; Elbr？chter, M.; Krock, B.; John, U.; Cembella, A. Azadinium spinosum gen. et sp. nov. (Dinophyceae) identified as a primary producer of azaspiracid toxins. Eur. J. Phycol. 2009, 44, 63–79.
[32]
Tillmann, U.; Elbr？chter, M.; John, U.; Krock, B.; Cembella, A. Azadinium obesum (Dinophyceae), a new nontoxic species in the genus that can produce azaspiracid toxins. Phycologia 2010, 49, 169–182, doi:10.2216/PH09-35.1.
[33]
Tillmann, U.; Elbr？chter, M.; John, U.; Krock, B. A new non-toxic species in the dinoflagellate genus Azadinium: A. poporum sp. nov. Eur. J. Phycol. 2011, 46, 74–87, doi:10.1080/09670262.2011.556753.
[34]
Tillmann, U.; Salas, R.; Gottschling, M.; Krock, B.; O’Driscoll, D.; Elbr？chter, M. Amphidoma languida sp. nov. (Dinophyceae) reveals a close relationship between Amphidoma and Azadinium. Protist 2012, 163, 701–719.
