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Seasonal dynamics of harmful algae in outer Oslofjorden monitored by microarray, qPCR, and microscopy

  • Molecular tools for monitoring Harmful Algal Blooms
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Abstract

Monitoring of marine microalgae is important to predict and manage harmful algal blooms. Microarray Detection of Toxic ALgae (MIDTAL) is an FP7-funded EU project aiming to establish a multi-species microarray as a tool to aid monitoring agencies. We tested the suitability of different prototype versions of the MIDTAL microarray for the monthly monitoring of a sampling station in outer Oslofjorden during a 1-year period. Microarray data from two different versions of the MIDTAL chip were compared to results from cell counts (several species) and quantitative real-time PCR (qPCR; only Pseudochattonella spp.). While results from generation 2.5 microarrays exhibited a high number of false positive signals, generation 3.3 microarray data generally correlated with microscopy and qPCR data, with three important limitations: (1) Pseudo-nitzschia cells were not reliably detected, possibly because cells were not sufficiently retained during filtration or lysed during the extraction, and because of low sensitivity of the probes; (2) in the case of samples with high concentrations of non-target species, the sensitivity of the arrays was decreased; (3) one occurrence of Alexandrium pseudogonyaulax was not detected due to a 1-bp mismatch with the genus probe represented on the microarray. In spite of these shortcomings our data demonstrate the overall progress made and the potential of the MIDTAL array. The case of Pseudochattonella — where two morphologically similar species impossible to separate by light microscopy were distinguished — in particular, underlines the added value of molecular methods such as microarrays in routine phytoplankton monitoring.

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Acknowledgments

We would like to thank Rita Amundsen, Tor Fredrik Holth, as well as the crew of the research vessel Trygve Braarud for their support during the monthly samplings, Shuhei Ota for EM analyses of the June 2010 samples, as well as the partners of MIDTAL for helpful discussions and support throughout the project. This work was funded by the EU’s seventh Framework Program (FP7-ENV-2007-1-MIDTAL-201724).

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Authors and Affiliations

  1. Marine Biology, Department of Biology, University of Oslo, P.O. Box 1066, Blindern, 0316, Oslo, Norway

    Simon M. Dittami, Vladyslava Hostyeva, Elianne Sirnæs Egge, Wenche Eikrem & Bente Edvardsen

  2. Station Biologique de Roscoff, UMR1739, Place Georges Teissier, 29680, Roscoff, France

    Simon M. Dittami

  3. Marine Biological Association of the United Kingdom, The Laboratory, Citadel Hill, Plymouth, PL1 2PB, UK

    Jessica U. Kegel

  4. Norwegian Institute for Water Research, Gaustadalléen 21, 0349, Oslo, Norway

    Wenche Eikrem

Authors
  1. Simon M. Dittami
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  2. Vladyslava Hostyeva
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  3. Elianne Sirnæs Egge
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  4. Jessica U. Kegel
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  5. Wenche Eikrem
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  6. Bente Edvardsen
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Correspondence to Simon M. Dittami or Bente Edvardsen.

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Responsible editor: Philippe Garrigues

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Table S1

List of probes represented on the MDITAL microarray generation 2.5. Probe sequences are not provided because the microarray is patent pending. Probes on generation 3.3 arrays have the same specificity and sequence, but have the suffix “_dT” was added to the probe name (DOC 123 kb)

Table S2

Estimations of detection limits for selected probes as determined by different partners of the MIDTAL project and published by Kegel et al. (2012) with modifications according to Edvardsen et al. (2012) for Dinophysis spp. (DOC 35 kb)

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Dittami, S.M., Hostyeva, V., Egge, E.S. et al. Seasonal dynamics of harmful algae in outer Oslofjorden monitored by microarray, qPCR, and microscopy. Environ Sci Pollut Res 20, 6719–6732 (2013). https://doi.org/10.1007/s11356-012-1392-0

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