Determination of optimal culture conditions for toxin production by a Prorocentrum lima complex strain with high diarrhetic shellfish toxins yield.

Diarrhetic shellfish poisoning (DSP) is caused by the consumption of shellfish contaminated by diarrhetic shellfish toxins (DSTs) such as okadaic acid (OA) and dinophysistoxins (DTXs) produced by some species of dinoflagellates. To prevent the occurrence of human intoxication cases, inspection of DSTs (OA and DTXs) in shellfish is important. An instrumental method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been recently employed in Japan for the monitoring of OA and DTXs in shellfish. For such analysis, reference materials (RMs) of OA and DTXs are essential. Demand for the reference materials, especially dinophysistoxin-1 (DTX1), is recently increasing in Japan. Production of the materials has been performed by mass cultivation of a dinoflagellate (Prorocentrum lima) strain that produces DTXs and OA, which indicates that the efficiency of production depends on the toxin production of the strain used. In this study, P. lima complex subclade 1e strain MIO12P was determined to be a high DTX1 producer among the three Japanese strains of the P. lima complex (subclades 1e, 1f, and 1i). It was clarified that the culture medium suitable for toxin production by strain MIO12P was metals mix SWII medium, and the optimal temperature and salinity for toxin production were 25 °C and salinity 30, respectively. The DTX1 yield (1265.3 ng ml-1) of strain MIO12P cultured under the conditions described above was the highest reported worldwide. Prorocentrum lima complex subclade 1e strain MIO12P is expected to be useful for the sustainable production of DTX1 as a source of RMs for chemical and biochemical methods in the future.

Diel vertical movements and feeding behaviour of blue humphead parrotfish Scarus ovifrons in a temperate reef of Japan.

The feeding ecology of scarinine parrotfishes on tropical coral reefs has received considerable attention in the past few decades; nonetheless, relatively few studies have been conducted in high-latitude reefs. Among the Indo-Pacific Scarus species, Scarus ovifrons is unique, being largely restricted to the warm temperate waters of Japan. Nonetheless, there is very little information available on the feeding ecology of this species. In this study, the authors used acoustic telemetry to detect the diel vertical movement patterns of S. ovifrons, video survey to detect its feeding depths and substrata and focal follow survey and genetic analysis to identify algae composition on the feeding scars at Kashiwajima Island, southwestern Japan (32° 46' N, 132° 38' E). Acoustic telemetry revealed that S. ovifrons spent most of its time in shallow water (<10 m) during the day and slept in deeper water (10-15 m) at night. Video and focal follow surveys revealed that most fishes of various sizes regularly took bites on epilithic algae and detrital materials on rocky substrata at depths of <10 m, but large fishes (>40 cm total length) sometimes took bites directly on live corals (Acropora solitaryensis) at the 5 m depth zone where live tabular corals dominated the benthos. Molecular phylogenetic analyses revealed that epilithic algae collected from feeding scars were mainly composed of Rhodophyta, and coralline algae were less often targeted. Overall, this study revealed that S. ovifrons feeds mostly at depths <10 m, and the feeding algae substrata of the species are similar to those of tropical coral reef parrotfishes.

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae).

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R>0.92 (p<0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (Lmax) and the semi-optimum light intensity range (Ls-opt) that resulted in the maximum growth rate (µmax) and ≥80% µmax values of the four species/phylotypes, respectively, were as follows: (1) the Lmax and Ls-opt of G. australes were 208µmol photons m-2s-1 and 91-422µmol photons m-2s-1, respectively; (2) those of G. scabrosus were 252 and 120-421µmol photons m-2s-1, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75-430µmol photons m-2s-1, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73-427µmol photons m-2s-1, respectively. All four Gambierdiscus species/phylotypes required approximately 10µmol photons m-2s-1 to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer's Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058-0.119m-1. The values 1700µmol photons m-2s-1, 500µmol photons m-2s-1, and 200µmol photons m-2s-1 were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of Lmax and Ls-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12-38m and 12-54m, 1-16m and 1-33m, and 0m and 0-16m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.