Molecular Phylogeny, Morphology, Growth and Toxicity of Three Benthic Dinoflagellates Ostreopsis sp. 9, Prorocentrum lima and Coolia monotis Developing in Strait of Gibraltar, Southwestern Mediterranean.

Few works have been carried out on benthic harmful algal blooms (BHAB) species in the southern Mediterranean and no data are available for the highly dynamic Strait of Gibraltar (western Mediterranean waters). For the first time, Ostreopsis sp. 9, Prorocentrum lima and Coolia monotis were isolated in this key region in terms of exchanges between the Atlantic Ocean and the Mediterranean and subject to intense maritime traffic. Ribotyping confirmed the morphological identification of these three dinoflagellates species. Monoclonal cultures were established and the maximum growth rate and cell yield were measured at a temperature of 24 °C and an irradiance of 90 µmol photons m-2 s-1, for each species: 0.26 ± 0.02 d-1 (8.75 × 103 cell mL-1 after 28 days) for Ostreopsis sp. 9, 0.21 ± 0.01 d-1 (49 × 103 cell mL-1 after 145 days) for P. lima and 0.21 ± 0.01 d-1 (10.02 × 103 cell mL-1 after 28 days) for C. monotis. Only P. lima was toxic with concentrations of okadaic acid and dinophysistoxin-1 measured in optimal growth conditions ranging from 6.4 pg cell-1 to 26.97 pg cell-1 and from 5.19 to 25.27 pg cell-1, respectively. The toxin content of this species varied in function of the growth phase. Temperature influenced the growth and toxin content of P. lima. Results suggest that future warming of Mediterranean coastal waters may lead to higher growth rates and to increases in cellular toxin levels in P. lima. Nitrate and ammonia affected the toxin content of P. lima but no clear trend was noted. In further studies, we have to isolate other BHAB species and strains from Strait of Gibraltar waters to obtain more insight into their diversity and toxicity.

Occurrence of three dominant epibenthic dinoflagellates (Ostreopsis spp., Coolia monotis and Prorocentrum lima) in relation to biotic substrates and environmental factors in a highly dynamic ecosystem, the Strait of Gibraltar (Southwestern Mediterranean).

No studies have been carried out on the benthic harmful algal blooms (BHABs) along the Strait of Gibraltar in the Mediterranean, and little is known about the diversity of blooming species. Here, epibenthic dinoflagellates were monitored at least biweekly over 18 months (May 2019-November 2020) in Oued Lihoud, Cap Malabata and Dalia on the thalli of five dominant macrophytes and in the water column. This is the first report on the seasonal distribution of BHAB species hosted by natural biotic substrates in the Strait of Gibraltar, which is known for high hydrodynamics, major entry of Atlantic waters and important maritime traffic. Three BHAB dinoflagellates were observed in the surveyed areas: Ostreopsis spp., Coolia monotis and Prorocentrum lima. The analysis of all data at the three sites showed that Dictyota dichotoma was the most favourable macroalgae host for these benthic dinoflagellates. The highest cell densities were observed in Cap Malabata for Ostreopsis spp. (2.7 × 105 cells/g fresh weight in September 2020), P. lima (4.57 × 104 cells/g FW in September 2020) and C. monotis (4.07 × 104 cells/g FW in June 2019). Phosphate and temperature were positively correlated to the abundances of the studied thermophilic BHAB species. In contrast, negative correlations were recorded with salinity, ammonium, nitrite, nitrate, DIN, nitrogen/phosphate ratio and suspended material, attesting of the complex relationships between environmental factors and BHAB species dynamic in each marine ecosystem. Toxin analyses of the natural phytoplankton assemblage during BHABs showed the presence of only lipophilic toxins, namely okadaic acid and dinophysistoxins produced by P. lima. These BHABs species have to be isolated to establish monoclonal cultures for ribotyping and ecophysiological investigations.

Seasonal variations of phytoplankton assemblages in relation to environmental factors in Mediterranean coastal waters of Morocco, a focus on HABs species.

Studies on phytoplankton and in particular Harmful Algal Blooms (HABs) species in southern Mediterranean waters are scarce. We performed from April 2008 to June 2009 weekly investigations on microphytoplankton community structure and abundance in two contrasted marine ecosystems located in the western Moroccan Mediterranean coast, M'diq Bay and Oued Laou Estuary. Simultaneously, we measured the main physico-chemical parameters. Globally, the two studied areas showed comparable values of the assessed abiotic environmental factors. Temperature and salinity followed seasonal variation with values ranging from 13.5 °C to 21.4 °C and 31 to 36.8, respectively. Average nutrient values in surface water ranged from 0.7 to 45.76 µM for dissolved inorganic nitrogen, 0.02-2.10 µM for PO4 and 0.23-17.46 µM for SiO4 in the study areas. A total of 92 taxa belonging to 8 taxonomic classes were found. The highest number of microphytoplankton abundance reached 1.2 × 106 cells L-1 with diatoms being the most abundant taxa. Factorial Discriminant Analysis (FDA) and Spearman correlation test showed a significant seasonal discrimination of dominant microphytoplankton species. These micro-organisms were associated with different environmental variables, in particular temperature and salinity. Numerous HABs species were encountered regularly along the year. Although Dinophysis species and Prorocentrum lima were present in both sites, no Lipophilic Shellfish Poisoning was detected for the analyzed bivalve mollusks. Domoic acid (DA), produced by toxic species of Pseudo-nitzschia was found with concentrations up to 18 µg DA g-1 in the smooth clam Callista chione. Data showed that the observed persistent and dramatic Paralytic Shellfish Poisoning (PSP) intoxication of mollusks resulted probably of Gymnodinium catenatum proliferations in both studied areas. Contrary to C. chione, the cockle Achanthocardia tuberculatum showed a permanent and extremely high toxicity level during the 15 months survey with up to 7545 µg Equivalent Saxitoxin kg-1 flesh (ten times higher than the sanitary threshold of 800 µg eqSTX Kg-1flesh). The present work highlights for the first time the dynamic of microphytoplankton including HABs species and their associated toxin accumulation in the commercially exploited shellfish in the southern western Mediterranean waters of Morocco. Furthermore, the acquired data will help us to improve the monitoring of HABs species and related toxins in these coastal marine systems.