Over the last fifteen years, blooms of the genus Ostreopsis have been reported more frequently and at higher abundances in the Mediterranean area. Ostreopsis cf. ovata is known to produce ovatoxins (OVTXs), structural analogues of palytoxin, which is one of the most potent non-polymeric toxins. However, the production of OVTXs is poorly characterized in situ. The present study focuses on toxin content and profile according to the bloom phase during summer 2017 in Villefranche-sur-Mer, France (NW Mediterranean Sea), depth (from 0.5 to 5 m) and three different macroalgal substrates of this epiphytic dinoflagellate (Padina pavonica, Dictyota spp. and Halopteris scoparia). Ovatoxin quantification of all samples was performed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The bloom started at the end of June and declined in mid-July, showing the typical seasonal pattern of the NW Mediterranean Sea area. The peak was observed on the 10 July with 1.8 × 106 cells/g FW and 1.7 × 104 cells/L for benthic and planktonic cells, respectively. Total toxin content of cells, collected using artificial substrates, increased during the exponential and stationary growth phases. After reaching a maximum concentration of 9.2 pg/cell on 18 July, toxin concentration decreased and remained stable from 25 July until the end of monitoring. A decreasing trend of the abundance and of the associated total toxin content was noted with depth. Finally, the decreasing order of maximal epiphytic concentration of O. cf. ovata was: Dictyota spp. (8.3 × 105 cells/g FW), H. scoparia (3.1 × 105 cells/g FW) and P. pavonica (1.6 × 105 cells/g FW). Interestingly, the highest OVTX quota was obtained in cells present on Halopteris scoparia, then on Dictyota spp. and Padina pavonica. This suggests that the nature of the macroalgal substrate influences both growth and toxin production of O. cf. ovata and further work will be required to understand the underlying mechanisms (e.g., competition for nutrition, pH or allelopathic interaction). However, the toxin profiles (i.e., the proportion of each ovatoxin analogue) were not affected by any of the studied parameters (bloom phase, depth, macroalgae or artificial substrates). Highlights► Ostreopsis cf. ovata bloomed from late June to August 2017 at Villefranche-sur-Mer, France, Mediterranean Sea. ► Ovatoxins contents were higher during the exponential and stationary growth phases. ► Highest abundance was on Dictyota spp. but highest toxin quota on Halopteris scoparia. ► Ovatoxin profile was not affected by the bloom phase, the depth or the macroalgae.

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Jézéquel. Effect of bacteria on growth and biochemical composition of two benthic diatoms Halamphora coffeaeformis and Entomoneis paludosa. Journal of Experimental Marine Biology and Ecology, Elsevier, 2017, 495, pp.65 -74 between Bacteroidetes (Flavobacteriia) and Proteobacteria (alpha-and gammaproteobacteria).After treatment with antibiotics, the residual bacterial community was ~37% Flavobacteriia

Even though HPLC-MS is commonly used to quantify the toxin content of Ostreopsis spp. cells, there is a need to develop easy-to-use toxicological tests to set thresholds during Ostreopsis spp. blooms. The crustacean Artemia has been widely used to evaluate the presence and toxicity of chemicals and biological contaminants and we anticipated that it could also be useful to test Ostreopsis spp. toxicity. Its relevance was first assessed by investigating the variability of the toxic effects among Ostreopsis spp. strains and throughout the dinoflagellate life cycle in combination with chemical analyses of the toxinic content by UHPLC-HRMS. After testing the toxicity of fractions prepared from Ostreopsis spp. cells, the known ova-and paly-toxins were not the only toxic metabolites to Artemia franciscana, indicating that other toxic compounds synthesized by Ostreopsis spp. still remain to be identified. To extend the bioassay to in situ monitoring, the toxicity of the benthic microalgal consortium was tested during a natural bloom of Ostreopsis cf. ovata in the NW Mediterranean Sea. The results highlight the accuracy and sensitivity of the ecotoxicological assay with Artemia franciscana to assess the toxicity of Ostreopsis spp. blooms. Highlights► Artemia bioassay is relevant to evaluate Ostreopsis spp. Toxicity ► Ostreopsis spp. produce other toxic compounds than known ovatoxins ► Artemia bioassay could be used for bio-guided fractionation approach