Rapid detection and quantification of the marine toxic algae, Alexandrium minutum, using a super-paramagnetic immunochromatographic strip test

The dinoflagellates of Alexandrium genus are known to be producers of paralytic shellfish toxins that regularly impact the shellfish aquaculture industry and fisheries. Accurate detection of Alexandrium including A. minutum is crucial for environmental monitoring and sanitary issues. In this study, we firstly developed a quantitative lateral flow immunoassay (LFIA) using super-paramagnetic nanobeads for A. minutum whole cells. This dipstick assay relies on two distinct monoclonal antibodies used in a sandwich format and directed against surface antigens of this organism. No sample preparation is required. Either frozen or live cells can be detected and quantified. The specificity and sensitivity are assessed by using phytoplankton culture and field samples spiked with a known amount of cultured A. minutum cells. This LFIA is shown to be highly specific for A. minutum and able to detect reproducibly 105 cells/L within 30 min. The test is applied to environmental samples already characterized by light microscopy counting. No significant difference is observed between the cell densities obtained by these two methods. This handy super-paramagnetic lateral flow immnunoassay biosensor can greatly assist water quality monitoring programs as well as ecological research.

Light and dark carbon uptake by Dinophysis species in comparison to other photosynthetic and heterotrophic dinoflagellates

The marine dinoflagellate genus Dinophysis includes species that are the causative agents of diarrhetic shellfish poisoning (DSP). Recent findings indicate that some Dinophysis species are mixotrophic, i.e. capable of both autotrophic and heterotrophic nutrition. We investigated inorganic (and organic) carbon uptake by several species of Dinophysis in the Light and dark using the 'single-cell C-14 method', and compared uptake rates with those of photosynthetic Ceratium species and heterotrophic dinoflagellates in the genus Protoperidinium. Experiments were conducted with water from the Gullmar Fjord and from the Koster Strait (Swedish west coast). Nutrient-enriched phytoplankton from surface water samples were concentrated (20 to 70 mu m) and incubated at in situ temperature under artificial light conditions with high concentrations of inorganic C-14 (1 mu Ci ml(-1)). Individual cells of each desired species were manually isolated under a microscope and transferred to scintillation vials. C. tripes showed net C-14 uptake only during light periods, whereas both C. lineatum and C. furca showed C-14 uptake in the Light as well as uptake (and sometimes losses) in the dark. Dinophysis species had similar carbon fixation rates in Light compared to Ceratium species. For D. acuminata and D. norvegica, net carbon uptake occurred in both Light and dark periods. D. acuta showed a loss of carbon in the dark in one experiment, but in another, dark C uptake was significantly higher than uptake in Light. When exposed to Light, C. furca, D. norvegica and D. acuta had high specific carbon uptake rates. Growth rates for the different species were calculated from C-14 uptake by the cells during the first hours of incubation in light. D. acuminata and D. norvegica had similar maximum growth rates, 0.59 and 0.63 d(-1) (mu); the maximum growth rate of D. acuta was lower (0.41 d(-1)). The positive dark carbon uptake by Dinophysis may suggest a mixotrophic mode of nutrition. In one experiment, both D. norvegica and D. acuta showed a significantly higher carbon uptake in a dark bottle than in a Light bottle, which would be consistent with uptake of C-14-labeled organic matter by D. norvegica and D. acuta. Demonstration of direct uptake of dissolved and particulate organic matter would provide conclusive evidence of mixotrophy and this will require the development of new protocols for measuring organic matter uptake applicable to Dinophysis in the natural assemblages.