Recurrent fish kills in the Spanish Alfacs Bay (NW Mediterranean) have been detected during winter seasons since 1994, and were attributed to an unarmored, ichthyotoxic, dinoflagellate, initially identified as Gyrodinium corsicum Paulmier, Berland, Billard, & Nezan. Several strains were isolated from the bay and their clonal cultures were compared by combined techniques, including light and electron microscopy, internal transcribed spacer and 5.8S rDNA nucleotide sequencing, and HPLC pigment analyses, together with studies of their photochemical performance, growth rates, and toxicity. Using phylogenetic analyses, all strains were identified as members of the genus Karlodinium, but they were separated into two genetically distinct groups. These groups, identified as Karlodinium veneficum (Ballantine) J. Larsen and K. armiger Bergholtz, Daugbjerg et. Moestrup, were also supported by the other techniques used. Detailed analyses of fine structural characteristics (including plug-like structures in amphiesma and a possible layer of semi-opaque material beneath the outer membrane) allowed discrimination of the mentioned two species. Specific differences in pigment patterns coincided with that expected for low-(K. veneficum) and high-light (K. armiger) adapted relatives. The higher photosynthetic efficiency of K. veneficum and the longer reactivation times of the PSII reaction centers observed for K. armiger were in agreement with this hypothesis. The two species differed in toxicity, but the strains used always induced mortality when incubated with bivalves, rotifers, and finfish. Compared with K. armiger, strains of K. veneficum yielded higher cell densities, but had lower growth rates.
Abstract. Some dinoflagellate species have shown different physiological responses to certain turbulent conditions. Here we investigate how two levels of turbulent kinetic energy dissipation rates (ε = 0.4 and 27 cm 2 s −3 ) affect the PSP toxins and ecdysal cyst dynamics of two bloom forming species, Alexandrium minutum and A. catenella. The most striking responses were observed at the high ε generated by an orbital shaker. In the cultures of the two species shaken for more than 4 days, the cellular GTX(1+4) toxin contents were significantly lower than in the still control cultures. In A. minutum this trend was also observed in the C(1+2) toxin content. For the two species, inhibition of ecdysal cyst production occurred during the period of exposure of the cultures to stirring (4 or more days) at any time during their growth curve. Recovery of cyst abundances was always observed when turbulence stopped. When shaking persisted for more than 4 days, the net growth rate significantly decreased in A. minutum (from 0.25±0.01 day −1 to 0.19±0.02 day −1 ) and the final cell numbers were lower (ca. 55.4%) than in the still control cultures. In A. catenella, the net growth rate was not markedly modified by turbulence although under long exposure to shaking, the cultures entered earlier in the stationary phase and the final cell numbers were significantly lower (ca. 23%) than in the control flasks.The described responses were not observed in the experiments performed at the low turbulence intensities with an orbital grid system, where the population development was favoured. In those conditions, cells appeared to escape from the zone of the influence of the grids and concentrated in calmer thin layers either at the top or at the bottom of the containers.This ecophysiological study provides new evidences about the sensitivity to high levels of small-scale turbulence by two life cycle related processes, toxin production and encystment, in dinoflagellates. This can contribute to the understanding of the dynamics of those organisms in nature.
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