BackgroundA dinoflagellate genus Ostreopsis is known as a potential producer of Palytoxin derivatives. Palytoxin is the most potent non-proteinaceous compound reported so far. There has been a growing number of reports on palytoxin-like poisonings in southern areas of Japan; however, the distribution of Ostreopsis has not been investigated so far. Morphological plasticity of Ostreopsis makes reliable microscopic identification difficult so the employment of molecular tools was desirable.Methods/Principal FindingIn total 223 clones were examined from samples mainly collected from southern areas of Japan. The D8–D10 region of the nuclear large subunit rDNA (D8–D10) was selected as a genetic marker and phylogenetic analyses were conducted. Although most of the clones were unable to be identified, there potentially 8 putative species established during this study. Among them, Ostreopsis sp. 1–5 did not belong to any known clade, and each of them formed its own clade. The dominant species was Ostreopsis sp. 1, which accounted for more than half of the clones and which was highly toxic and only distributed along the Japanese coast. Comparisons between the D8–D10 and the Internal Transcribed Spacer (ITS) region of the nuclear rDNA, which has widely been used for phylogenetic/phylogeographic studies in Ostreopsis, revealed that the D8–D10 was less variable than the ITS, making consistent and reliable phylogenetic reconstruction possible.Conclusions/SignificanceThis study unveiled a surprisingly diverse and widespread distribution of Japanese Ostreopsis. Further study will be required to better understand the phylogeography of the genus. Our results posed the urgent need for the development of the early detection/warning systems for Ostreopsis, particularly for the widely distributed and strongly toxic Ostreopsis sp. 1. The D8–D10 marker will be suitable for these purposes.

BackgroundThe marine epiphytic dinoflagellate genus Gambierdiscus produce toxins that cause ciguatera fish poisoning (CFP): one of the most significant seafood-borne illnesses associated with fish consumption worldwide. So far, occurrences of CFP incidents in Japan have been mainly reported in subtropical areas. A previous phylogeographic study of Japanese Gambierdiscus revealed the existence of two distinct phylotypes: Gambierdiscus sp. type 1 from subtropical and Gambierdiscus sp. type 2 from temperate areas. However, details of the genetic diversity and distribution for Japanese Gambierdiscus are still unclear, because a comprehensive investigation has not been conducted yet.Methods/Principal FindingA total of 248 strains were examined from samples mainly collected from western and southern coastal areas of Japan during 2006–2011. The SSU rDNA, the LSU rDNA D8–D10 and the ITS region were selected as genetic markers and phylogenetic analyses were conducted. The genetic diversity of Japanese Gambierdiscus was high since five species/phylotypes were detected: including two reported phylotypes (Gambierdiscus sp. type 1 and Gambierdiscus sp. type 2), two species of Gambierdiscus (G. australes and G. cf. yasumotoi) and a hitherto unreported phylotype Gambierdiscus sp. type 3. The distributions of type 3 and G. cf. yasumotoi were restricted to the temperate and the subtropical area, respectively. On the other hand, type 1, type 2 and G. australes occurred from the subtropical to the temperate area, with a tendency that type 1 and G. australes were dominant in the subtropical area, whereas type 2 was dominant in the temperate area. By using mouse bioassay, type 1, type 3 and G. australes exhibited mouse toxicities.Conclusions/SignificanceThis study revealed a surprising diversity of Japanese Gambierdiscus and the distribution of five species/phylotypes displayed clear geographical patterns in Japanese coastal areas. The SSU rDNA and the LSU rDNA D8–D10 as genetic markers are recommended for further use.

No abstract

Benthic dinoflagellates of the genus Ostreopsis produce palytoxin (PTX)-like compounds. The worldwide distributed Ostreopsis ovata/O. cf. ovata is potentially responsible for outbreaks of human health problems around the coasts of tropical, subtropical, and temperate regions. The present study examined growth responses of an O. cf. ovata strain s0662 collected from coastal waters of Japan with 35 different combinations of temperature (15-35°C) and salinity (20-40) and discusses the bloom dynamics of the organism in Japanese coastal environments. The O. cf. ovata strain s0662 tolerated a wide range of temperature (17.5-35°C) and salinity (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). Results of a two-way ANOVA showed significant effects of temperature-salinity interaction on growth rates and biomass yields of the O. cf. ovata strain (F(24,70) > 127, P < 0.001). The strain showed a maximal growth rate (1.03 divisions day -1 ) and biomass yield (240 relative fluorescence) at temperature 25°C and salinity 30. The high growth rates of over 1.0 division day -1 were obtained in conditions of temperature 25-30°C and salinity 30-35, which indicates that strain s0662 prefers high temperature and salinity conditions. The growth rates of O. cf. ovata under the optimal conditions were higher than those of other benthic toxic-dinoflagellates, Coolia monotis, Gambierdiscus toxicus, and Prorocentrum lima (Dinophyceae) previously reported. Taken together, we suggest that O. cf. ovata is able to grow faster than the other benthic dinoflagellates in waters of high temperature and salinity. The physiological feature probably confers an ecological advantage on O. cf. ovata in the bloom development during warmer seasons in Japan and may be responsible for outbreaks of PTX-like poisoning in the region especially during the warmer seasons.

Ciguatera fish poisoning (CFP) is caused by toxins originating from an epiphytic/benthic dinoflagellate of the genus Gambierdiscus. In Japan, CFP cases have been increasingly reported not only in subtropical areas but also in temperate areas. It is therefore important to study Gambierdiscus cell occurrences, cell densities, and population dynamics to address CFP outbreaks in Japan. This study assessed the densities in Japanese shallow waters (0.1-3 m depths) and revealed that the densities were lower than those in tropical and subtropical areas worldwide. In the shallow waters of Tosa Bay, a Japanese temperate area, population dynamics of Gambierdiscus cells were assessed monthly between 2007 and 2013. Gambierdiscus did not show substrate preferences for macroalgal species. The cell densities in the area ranged from 0 to 232.2 cells g −1 wet weight algae. The average cell densities in spring, summer, autumn, and winter were 0.1±0.4, 0.9±2.6, 4.0±20.6, and 0.4±1.4 cells g −1 wet weight algae, respectively. The cell densities in summer and autumn were not significantly different (p>0.05), whereas those in summer and autumn were significantly higher than those in spring and winter (p<0.01). A significant positive correlation between cell densities and sea surface temperatures (SSTs) was observed (r s =0.21, p<0.001), while a significant negative correlation between cell densities and salinity was recognized (r s =−0.18, p<0.001). These results suggest that cell densities of Gambierdiscus in Japanese temperate shallow waters increase in summer and autumn when the SST is high and salinity is moderately low.