Estimation of 1999-2004 dietary daily intake of PCDDs, PCDFs and dioxin-like PCBs by a total diet study in metropolitan Tokyo, Japan

A study of the dietary intake of dioxins, consisting of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) through foods retailed in the metropolitan Tokyo area from 1999 to 2004 was carried out by the total diet-market basket method on the basis of food classification (14 groups) and the data on food consumption in the Tokyo region obtained from the Japan Nutrition Survey. The daily intake of dioxins per kg of body weight for a 50 kg average adult body was 2.18 pg TEQ/kg/day in 1999, 1.87 pg TEQ/kg/day in 2000, 1.25 pg TEQ/kg/day in 2001, 1.60 pg TEQ/kg/day in 2002 and 2003 and 1.55 pg TEQ/kg/day in 2004, respectively. These amounts were less than the tolerable daily intake (TDI) of 4 pg TEQ/kg/day for dioxins established in Japan. The dioxins taken daily through fish and shellfish (group 10) accounted for more than 50% of sum WHO-TEQs. In addition, more than 90% of the daily intake of dioxins was taken through fish and shellfish (group 10), meat and eggs (group 11), milk and dairy products (group 12). Also, this study clearly showed that the ratio of dioxin-like PCBs in the daily intake of dioxins was increasing yearly because the reduction rate of dioxin-like PCBs was lower than that of PCDDs and PCDFs in foods.     

Molecular taxonomy of vestimentiferans of the western Pacific,and their phylogenetic relationship to species of the eastern Pacific III. Alaysia-like vestimentiferans and relationships among families

The phylogenetic status of Alaysia-like vestimentiferans, which were collected at eight sites in the western Pacific, was analyzed on the basis of the nucleotide sequence of part of a mitochondrial gene for cytochrome oxidase I. The 123 individuals analyzed were tentatively classified into four species, which inhabit, respectively, seep areas off the central part of the Japanese mainland, hydrothermal vent fields in the Mid-Okinawa Trough, hydrothermal vent fields in the southern Okinawa Trough, and both seep and vent sites in the Bismarck Sea. From six additional lamellibrachiid specimens from two sites in the Bismarck Sea, two new tentative species were recognized, namely, one from a seep area off Papua New Guinea and another from the hydrothermal vent field at the DESMOS site in the Manus Basin. Phylogenetic analysis using all available sequences of six vestimentiferan families and Alaysia-like vestimentiferans revealed that vestimentiferans could be divided into three groups, namely, lamellibrachiids, escarpiids, and others, even though the monophyly of the third group was supported by only low bootstrap probabilities. Within the first group, the earliest divergence of a tentative lamellibrachiid species from the DESMOS site was apparent. All Alaysia-like vestimentiferans formed a monophyletic group with Arcovestia ivanovi from the Manus Basin, and it is suggested that this group might be derived from vestimentiferans that are endemic to the hydrothermal sites in the Eastern Pacific Rise.     

Evolutionary relationships of deep-sea mussels inferred by mitochondrial DNA sequences

In order to elucidate the evolutionary process of deep-sea Bathymodiolus mussels, we investigated the phylogenetic relationships of 16 species worldwide by analyzing nucleotide sequences of the mitochondrial COI and ND4 genes. Deep-sea mussels were clustered into three groups by basal trichotomous divergence. The first was composed of four species found in Japanese waters and one species from the Gulf of Mexico, which contain methanotrophic endosymbiotic bacteria. The second included nine species distributed in the West and East Pacific, Indian, and Atlantic Oceans. Members of the second group were trichotomously divided into the Indo-West Pacific, Atlantic, and East Pacific subclusters. The Indo-West Pacific subcluster was composed of three very closely related species with mutual genetic distances at the intraspecific level (av. 0.019 in COI and 0.009 in ND4 relative to av. 0.156 in COI and 0.265 in ND4 among Bathymodiolus species other than Cluster A species), suggesting some gene flow among these species. The third consisted of two West Pacific species. Species in the second and third groups contain mainly thioautotrophic endosymbionts, including some species harboring both methanotrophs and thioautotrophs.     

Phylogenetic relationships of deep-sea mussels of the genus<Emphasis Type="Italic"> Bathymodiolus</Emphasis> (Bivalvia: Mytilidae)

We examined phylogenetic relationships among three Bathymodiolus species in Japanese waters and Bathymodiolus spp. from the Manus Basin by two different approaches. Two-dimensional gel electrophoresis allowed us to compare 263–407 (average=318) proteins, giving comprehensive information on genetic distances among the species. The neighbor-joining tree presented two clusters: (1) B. japonicus and B. platifrons and (2) B. septemdierum and B. sp. Members of the first cluster contain methanotrophic endosymbiotic bacteria and members of the second cluster contain thioautotrophic endosymbionts. DNA sequencing of a fragment (415 bp) of mitochondrial cytochrome c oxidase subunit I (COI) provided a neighbor-joining tree with the same topology as that derived from protein analysis. Inspection of intraspecific variation in COI in B. japonicus and B. platifrons revealed no genetic differentiation between mussel populations of either species from cold-water seeps versus hydrothermal vents, suggesting high adaptability of these Bathymodiolus species to deep-sea chemosynthetic environments. Our results indicated genetic exchanges between mussels from distant localities, suggesting that a limited dispersal capability of the larvae is not the likely factor leading to speciation events in these Bathymodiolus species.Communicated by T. Ikeda, Hakodate     

Identification and significance of pyrochlorins in fecal pellets of the marine malacostracan crustaceans Heptacarpus rectirostris and Palaemon pacificus

The chlorophyll derivatives, especially pyrochlorins, from macrobentho-pelagic fecal pellets were separated and identified using reversed-phase high-per-formance liquid chromatography-mass spectrometry (HPLC-MS). HPLC-frit-fast atom bombardment (FAB)-MS analysis of fecal pellet extracts was performed, and the ions at m/z (mass-to-charge ratio), 534 [M]⁺, 827 [M+H]⁺ and 812 [M]⁺ as base peaks contirm the presence of pyropheophorbide a, pyropheophytin b and pyropheophytin a, respectively. Identification of pyrochlorins in fecal pellets suggests that decarbomethoxylation of chlorophylls can occur during bentho-pelagic grazing and that pyrochlorins in sedimentary environments may be derived in part from fecal pellets.     

Occurrence of a crinotoxin and hemagglutinin in the skin mucus of the moray eel Lycodontis nudivomer

The tropical Indo-Pacific moray eel Lycodontis nudivomer possesses a mucous skin secretion which shows hemolytic, toxic and hemagglutinating activities. The active components were extracted from the mucous secretion with 0.02 M phosphate buffer (pH 7.5) and purified by DEAE-cellulose and Sephadex G-200 column chromatography. The component(s) responsible for the hemolytic-toxic activities differ(s) from that (those) causing the hemagglutinating activity. The hemolysin was unstable in the presence of heat, acidic and alkaline media, and several organic solvents. It was undialyzable throught a cellophane membrane, and showed an absorption maxium at 280 nm in the phosphate buffer (pH 7.5). The hemolytic activity was completely lost when treated with trypsin, indicating that the hemolysin is a protein. Histochemical examination on the epidermis of L. nudivomer revealed the presence of clubbed peculiar cells, in addition to common mucous cells, which we speculate produced the hemolytic-toxic component(s). Eight other species of morays were found to possess clubbed peculiar cells similar to those of L. nudivomer in their epidermis. There are marked differences, however, in the abundance of these cells among these muraenids.     

Bioaccumulation of mercury in a vestimentiferan worm living in Kagoshima Bay,Japan

The present study reports on the mercury concentrations of the vestimentiferan worm, Lamellibrachia satsuma, (Annelida: Pogonophora) found near hydrothermal vents at a depth of 80-100 m in the northern parts of Kagoshima Bay. The vestimentiferan worms had total mercury concentrations of 238 ng/g in the anterior muscle of the body and 164 ng/g in the posterior trophosome. Methylmercury constituted only 7.6% of total mercury detected anteriorly and 16.3% posteriorly. The mean total mercury concentration in filtrated ambient seawater of the worm habitat was 1.1 ng/l. The worm should accumulate mercury in seawater by a one-step into the anterior and posterior parts as 2.2 x 10(%) and 1.5 x 10(5) times those of the filtered ambient seawater, respectively. The bioaccumulation factor of mercury by the worms with only their respiration would be actually larger than that by other marine animals through food webs. The high bioaccumulation factor of mercury in the worms suggest the following two possibilities: (i) the biological half-life of organomercury in the worm could be exceptionally long; or (ii) the lifetime of vestimentiferan worms examined in the present study could be extremely long. Various metals in one specimen of the worm were analyzed by using ICP-MS, and then gold as well as silver were detected in the worm. Gold was detected for the first time from marine animals.     

Variation in CO2 assimilation rate induced by simulated dew waters with different sources of hydroxyl radical (*OH) on the needle surfaces of Japanese red pine (Pinus densifora Sieb. et Zucc.)

The hydroxyl radical (*OH) is generated in polluted dew on the needle surfaces of Japanese red pine (Pinus densiflora Sieb. et Zucc.). This free radical, which is a potent oxidant, is assumed to be a cause of ecophysiological disorders of declining trees on the urban-facing side of Mt. Gokurakuji, western Japan. Mists of *OH-generating N(III) (HNO2 and NO2-) and HOOH + Fe + oxalate solutions (50 and 100 microM, pH 5.1-5.4) simulating the dew water were applied to the foliage of pine seedlings grown in open-top chambers in the early morning. Needles treated with 100 microM N(III) tended to have a greater maximum CO2 assimilation rate (Amax), a greater stomatal conductance (g(s)) and a greater needle nitrogen content (Nneedle), suggesting that N(III) mist acts as a fertilizer rather than as a phytotoxin. On the other hand, needles treated with 100 microM HOOH + Fe + oxalate solution showed the smallest Amax, g(s), and Nneedle, suggesting that the combination of HOOH + Fe + oxalate caused a decrease in needle productivity. The effects of HOOH + Fe + oxalate mist on pine needles were very similar to the symptoms of declining trees at Mt. Gokurakuji.