Occurrence of Prorocentrum lima,a DSP toxin-producing species from the Atlantic coast of Canada

A species of Prorocentrum (Dinophyta, Prorocentrales), isolated from a phytoplankton net sample from the Atlantic coast of Nova Scotia, has been brought into unialgal culture. The sample was collected at an aquaculture site immediately following an incident of diarrhetic shellfish poisoning (DSP) due to the consumption of contaminated mussels. This clonal isolate has been identified as P. lima, based on its morphological characteristics. Analysis of the culture extract, using high performance liquid chromatography (HPLC) with fluorescence detection, indicated the presence of the DSP toxins, okadaic acid (OA) and dinophysistoxin-1 (DTX-1).     

Genetic identification of fish eggs collected in Sendai Bay and off Johban,Japan

Mitochondrial DNA typing of 52 fish eggs collected in Sendai Bay and off Johban revealed ten haplotypes of the 225–251 bp region of 16S ribosomal DNA. Comparing with known sequences of fishes, we identified them as Trachurus japonicus (Carangidae), Pagrus major, Evynnis japonica (Sparidae), Paralichthys olivaceus, Pseudorhombus sp. (Paralichthidae), Pennahia argentata (Sciaenidae) and Platycephalus sp. (Platycephalidae). The occurrence of Trachurus japonicus and sparid eggs in the study area indicates significant extensions to the ranges of their spawning sites northward. Our method is useful for identifying fish eggs that are difficult to diagnose by morphology, and to uncover cryptic fish egg assemblages in the field.     

DSP toxin production de novo in cultures of Dinophysis acuminata (Dinophyceae) from North America

For decades, many aspects of Dinophysis biology have remained intractable due to our inability to maintain these organisms in laboratory cultures. Recent breakthroughs in culture methods have opened the door for detailed investigations of these important algae. Here, for the first time, we demonstrate toxin production in cultures of North American Dinophysis acuminata, isolated from Woods Hole, MA. These findings show that, despite the rarity of Dinophysis-related DSP events in North America, D. acuminata from this area has the ability to produce DSP toxins just as it does in other parts of the world where this species is a major cause of DSP toxicity. In our cultures, D. acuminata cells were observed feeding on Myrionecta rubra using a peduncle. Culture extracts were analyzed using LC–MS/MS, providing unequivocal evidence for the toxin DTX1 in the Dinophysis cultures. In addition, a significant amount of an okadaic acid diol ester, OA-D8, was detected. These results suggest that this Dinophysis isolate stores much of its OA as a diol ester. Also, toxin PTX-2 and a hydroxylated PTX-2 with identical fragmentation mass spectrum to that of PTX-11, but with a different retention time, were detected in this D. acuminata culture. This demonstration of toxin production in cultured North American Dinophysis sets the stage for more detailed studies investigating the causes of geographic differences in toxicity. It is now clear that North American Dinophysis have the ability to produce DSP toxins even though they only rarely cause toxic DSP events in nature. This may reflect environmental conditions that might induce or repress toxin production, genetic differences that cause modifications in toxin gene expression, or physiological and biochemical differences in prey species.     

Contribution to toxicity assessment of <Emphasis Type="BoldItalic">Dinophysis acuminata</Emphasis> (Dinophyceae)

Blooms of Dinophysis in French coastal waters are implicated in most bans on marketing commercial bivalves. However, the relation between Dinophysis cell density and shellfish toxicity is not always consistent. Discrepancies may be due to the simple fact that it is nearly impossible to compare an integral over a few days (shellfish toxin content) and water samples. Furthermore, it seems that cells may have a variable specific toxicity. This work focuses on the variability in cell toxicity taking into account recent findings and using liquid chromatography coupled to mass spectrometry with an ion trap and electrospray interface. Esterified analogues of okadaic acid (DTX-4 and diol-esters) have been identified in cultures of Prorocentrum lima, another okadaic acid producer. These analogues are inactive on some protein phosphatases, contrary to okadaic acid, and seem to protect the cell from harmful effects by the toxin and to be enzymatically hydrolyzed during cell lysis. In order to document specific toxicity and to validate the presence of these analogues, D. acuminata concentrates were subjected to two separate heating and freeze/thaw procedures, respectively inhibiting or promoting hydrolysis. This paper reports on the high variability of D. acuminata specific toxicity and the presence of esters found in half of the samples only.     

Larval and juvenile Polymetme elongata (Stomiiformes: Phosichthyidae) collected from Suruga Bay and offshore waters, Japan

Two larvae [17.4 mm standard length: SL (postflexion stage)] and 26.1 mm SL (transformation stage)] and a juvenile (31.7 mm SL) of a phosichthyid, Polymetme elongata, from Suruga Bay and offshore waters, central Japan, are described. These specimens had an elongate body with relatively short preanal length (53–63% SL), long anal fin base (2.6–3.4 times dorsal fin base length), and anal fin origin below dorsal fin base, and were further characterized by a blackish flap on each eye and internal clusters of melanophores (e.g., along caudal myosepta around midlateral line and on ventral margin of caudal peduncle). The short preanal length and larval melanophore pattern were very similar to those of another phosichthyid, Yarrella blackfordi, from the Atlantic Ocean.     

Occurrence of okadaic acid,a major diarrheic shellfish toxin,in natural populations ofDinophysis spp. from the eastern coast of North America

Okadaic acid, one of the principal toxin components implicated in cases of diarrheic shellfish poisoning (DSP), was identified for the first time in natural phytoplankton assemblages from North American waters. During periods in late summer when significant quantities of okadaic acid were detected in net haul samples in the lower estuary and Gulf of St Lawrence in eastern Canada, the phytoplankton community consistently contained species of the dinoflagellate genusDinophysis Ehrenberg. The presence of okadaic acid was detected by screening dinoflagellate extracts with an enzyme-linked immunological assay (ELISA); positive results were confirmed by reverse-phase high-performance liquid chromatography (HPLC) separation, followed by fluorescence detection. Okadaic acid was only found in phytoplankton samples in which the photosynthetic dinophysoid speciesD. norvegica andD. acuminata were prominent; blooms of the related heterotrophic speciesD. rotundata exhibited no trace of okadaic acid, nor other suspected DSP components.     

Okadaic acid accumulation in macrofilter feeders subjected to natural blooms of Dinophysis acuminata

Thermaikos Gulf is a eutrophic area located in the Northwestern part of the Aegean Sea in the Eastern Mediterranean. Interspecific differences among various filter feeders in their ability to accumulate okadaic acid, were observed during natural blooms of Dinophysis acuminata in the gulf. Okadaic acid analyses by high performance liquid chromatography (HPLC) were performed on benthic specimens and D. acuminata cell densities and cell toxin content were estimated in water samples. Seven filter feeding species were collected in the gulf during two DSP outbreaks in May 2003 and March 2004. The various species showed a different potential to accumulate okadaic acid in their tissues. The highest concentrations were found in the mussel populations (Mytilus galloprovincialis and Modiolus barbatus), while among the non-bivalve filter feeders, ascidians were the main accumulators of okadaic acid. The rest of shellfish populations (Flexopecten proteus, Chlamys varia and Venus verrucosa) were found to contain toxins only during 2004, when D. acuminata densities were found above 10000 cells l⁻¹. M. galloprovincialis was proved to be the most appropriate indicator for a safe warning of okadaic acid contamination in Thermaikos Gulf.     

Phytoplankton detection and DSP toxicity: methodological considerations

Two different phytoplankton sampling methods (bottle and net sampling) were used to evaluate the concentration of toxicDinophysis species in seawater and their correlation to mussel toxicity, assessed by mouse bioassay.Dinophysis concentration in net samples revealed the higher correlation to mussel toxicity (r=0.75,p<0.01). Net sampling therefore seems more suitable for the detection of low abundance species likeDinophysis characterised by vertical aggregations at different depths in the water column.     

Fauna of Sipunculans (Sipuncula) of Vostok Bay, Sea of Japan

Fauna and systematics of sipunculans inhabiting Vostok Bay, Sea of Japan, are discussed. A revision of the local species of sipunculans is performed for the first time. On the basis of study of outer and inner morphology, some species are reduced to synonyms. Extended diagnoses are proposed for all registered species. An identification key is presented for the sipunculans of Vostok Bay.     

Distribution and Recent Reduction of Gelidium Beds in Toyama Bay, Japan

The distribution and recent reduction of Gelidium beds, i.e. mat-like beds dominated by the agarophyte G. elegans Kützing in Toyama Bay (Sea of Japan), in which 95% of the coastline is protected artificially, are reported. Gelidium beds were common in shallow waters (usually < 10 m deep); most of the large beds (> 1 ha) were restricted to the inner coasts of the bay. In calm and eutrophic areas, however, G. elegans was heavily colonized by epiphytes. In the last decade, two beds were buried in situ and beds in their vicinity were damaged by the stagnation of coastal water and/or sedimentation by silts which accompanied land reclamation. At the other two beds monitored since 1988, Gelidium declined a few times but most prominently in 1998, when episodic long summer rain was recorded. This is the first report, not only on the current status of Gelidium beds other than for the central Pacific Coast of Honshu in Japan, but also concerning reduction of the beds caused by both anthropogenic and natural events.     

Vertical distribution and seasonal variation of eelgrass beds in Iwachi Bay,Izu Peninsula,Japan

The vertical distribution and population structure of eelgrass beds were surveyed in Iwachi Bay, along the Pacific coast of central Japan. Samplings were conducted from May through November 1977 by SCUBA. Eelgrass was distributed between 3 and 11 m in depth. The relative light intensity at 12 m depth was 11% at the lower range. The highest population density was 290 shoots/m² in September and the fresh weight of biomass was 888 g/m² in July at 7 m depth. The maximum mean leaf area index was about 3 at 10 m depth in July. The ratio of reproductive shoots to the total shoots was about 36% at 7 m depth in June. Eelgrass showed good growth at 7–10 m depth, which is comparatively deeper than other eelgrass habitations. The high values of water transparency and sunshine duration, as well as solar radiation compared with other localities was believed to contribute to the growth of eelgrass in deeper waters in Iwachi Bay.     

Three new species ofCrinipellis found in Iriomote Island, southwestern Japan, and central Honshu, Japan

Three new species ofCrinipellis (Agaricales, Basidiomycetes) in Japan are described and illustrated:Crinipellis canescens sp. nov., forming small, canescent basidiomata, was found on a dead fallen twig in the subtropical laurelleaved forest of Iriomote Island (southwestern Japan);Crinipellis corvina sp. nov., forming fibrillose-strigose, puriplish black hairs enveloping the whole basidioma, was found on bark ofTorreya nucifera in Mt. Takao, Tokyo;Crinipellis nigricaulis sp. nov., having a reddish brown pileus with a minute, black papilla and a blackish stipe accompanied by blackish, branched rhizomorphs, was found on leaf litter in the lowland forest of Kanagawa.     

Growth and Longevity of the Gastropod Nucella heyseana in Vostok Bay,Sea of Japan

Features of the morphology, longevity, and growth of the gastropod mollusk Nucella heyseana collected in different seasons in 2000–2001 in Vostok Bay (Sea of Japan) were investigated. Shell surface features such as grooves and spurs of the axial sculpture have proven to be unhelpful in the estimation of individual age and growth rate of this species. Concentric rings on the outside of the operculum formed yearly in the cold season are useful for these purposes. The longevity of N. heyseana in southern Primorye is 10 years; this value is similar to the life span of mollusks from the southern Kuril Islands. Local differences in longevity, growth allometry, and age changes in the body size and mass of N. heyseana in relation to habitat conditions are discussed.     

Development of Expressed Sequence Tags from the Bay Scallop, Argopecten irradians irradians

The bay scallop, Argopecten irradians irradians, introduced from North America, has become one of the most important aquaculture species in China. Inan effort to identify scallop genes involved in host defense, a high-quality cDNA library was constructed from whole body tissues of the bay scallop. A total of 5828 successful sequencing reactions yielded 4995 expressed sequence tags (ESTs) longer than 100 bp. Cluster and assembly analyses of the ESTs identified 637 contigs (consisting of 2853 sequences) and 2142 singletons, totaling 2779 unique sequences. Basic Local Alignment Search Tool (BLAST) analysis showed that the majority (73%) of the unique sequences had no significant homology (E-value ≤ 0.005) to sequences in GenBank. Among the 748 sequences with significant GenBank matches, 160 (21.4%) were for genes related to metabolism, 131 (17.5%) for cell/organism defense, 124 (16.6%) for gene/protein expression, 83 (11.1%) for cell structure/motility, 70 (9.4%) for cell signaling/communication, 17 (2.3%) for cell division, and 163 (21.8%) matched to genes of unknown functions. The list of host-defense genes included many genes with known and important roles in innate defense such as lectins, defensins, proteases, protease inhibitors, heat shock proteins, antioxidants, and Toll-like receptors. The study provides a significant number of ESTs for gene discovery and candidate genes for studying host defense in scallops and other molluscs.     

Characterization of fatty acid composition in healthy and bleached corals from Okinawa, Japan

Under bleaching conditions, corals lose their symbiotic zooxanthellae, and thus, the ability to synthesize fatty acids (FAs) from photosynthetically derived carbon. This study investigated the lipid content and FA composition in healthy and bleached corals from the Odo reef flat in Okinawa, southern Japan, following a bleaching event. It was hypothesized that the FA composition and abundance would change as algae are lost or die, and possibly microbial abundance would increase in corals as a consequence of bleaching. The lipid content and FA composition of three healthy coral species (Pavona frondifera, Acropora pulchra, and Goniastrea aspera) and of partially bleached and completely bleached colonies of P. frondifera were examined. The FA composition did not differ among healthy corals, but differed significantly among healthy, partially bleached, and completely bleached specimens of P. frondifera. Completely bleached corals contained significantly lower lipid and total FA content, as well as lower relative amounts of polyunsaturated FAs and higher relative amounts of saturated FAs, than healthy and partially bleached corals. Furthermore, there was a significantly higher relative concentration of monounsaturated FAs and odd-numbered branched FAs in completely bleached corals, indicating an increase in bacterial colonization in the bleached corals.     

Photosynthetic and respiratory responses of Gracilaria vermiculophylla (Ohmi) Papenfuss collected from Kumamoto, Shizuoka and Iwate, Japan

The photosynthetic and respiratory responses to irradiance, salinity and temperature of the red alga, Gracilaria vermiculophylla, collected from Kumamoto, Shizuoka and Iwate in Japan were studied using an electronic Dissolved Oxygen sensor. The parameters derived from the photosynthesis versus irradiance relationship indicated the potential to acclimate to broad irradiance variations in all of the populations of G. vermiculophylla collected from these three sites. In addition, the light-saturated photosynthesis rate (P _max) and the dark respiration rate of all populations increased with increasing temperature up to 20–30°C, while the P _max decreased at 35°C. All populations also showed a broad variation of photosynthetic responses to salinity changes in the range from 10 to 30 psu. On the other hand, the population from Iwate showed high photosynthetic efficiency, especially in the temperature range of 5–10°C, and showed low values of saturation irradiance compared to the populations from Shizuoka and Kumamoto. These results suggest that there is greater potential to acclimate to low irradiance and low temperature in the population from Iwate compared to those from the Shizuoka and Kumamoto populations. However, the P _max of the populations from Iwate and Shizuoka was reached at 20°C and 25°C, respectively, while the Kumamoto population reached P _max at 30°C. This implies that the latter population has greater potential to tolerate higher temperatures than the former. Such characteristics in photosynthesis and respiration of G. vermiculophylla collected from the three locations probably indicate an acclimation to prevailing environmental conditions in their respective habitats.     

Diet and prey selection in larval and juvenile Japanese anchovy Engraulisjaponicus in Ariake Bay, Japan

We studied the diet of larval and juvenile Japanese anchovy Engraulis japonicus in the upper Ariake Bay, Japan. Diet was analyzed by examining the digestive tracts; feeding intensity, proportion of empty guts, and prey selectivity were calculated. Anchovy density was negatively influenced by temperature and positively by salinity and prey density. Diet was dominated by Acartia omorii, which was positively selected with two other copepods, Calanus sinicus and Pseudodiaptomus marinus. In contrast, Oithona davisae was highly dominant in the environment but was absent in anchovy guts; thus, this copepod was negatively selected, with two others, Tortanus derjugini and Sinocalanus sinensis. Overall, larger prey were positively selected and smaller ones were negatively selected; value of electivity index correlated negatively with prey size. Larvae [<18 mm of standard length (SL)] showed significantly lower feeding intensities and higher rates of empty guts than juveniles (≥18 mm SL). In juveniles, feeding intensity increased steadily as the fish grew in size, with a corresponding reduction in empty guts. Feeding intensity correlated positively and empty gut correlated negatively with fish size. We suggest that larger prey are important diets for postlarval Japanese anchovy in Ariake Bay.