Distribution of hydrocarbons in seabed sediments derived from tsunami-spilled oil in Kesennuma Bay, Japan.

As a result of the 2011 Great East Japan earthquake, about 11,000kL of fuel oil was spilled into Kesennuma Bay, Miyagi Prefecture. This oil either accumulated in seabed sediments or was burned in a marine fire on the sea surface. We investigated spatial and temporal variations in the concentrations of oil and hydrocarbons. The maximum concentrations of n-hexane extract (NHE), total petroleum hydrocarbon (TPH), and polycyclic aromatic hydrocarbons (PAHs) in the sediments in 2012 were 8000, 1200, and 16.2mg/kg (DW), respectively. NHE and TPH concentrations were high in the inner and west bay, and PAHs concentrations were high in the east bay. NHE and PAH concentrations didn't change; however, TPH concentrations decreased significantly with time. The total NHE in the sediment across the whole bay was estimated at 1685kL and there was still about 13% of the original amount in the sediment in December 2014.

High spatial resolution analysis of the distribution of sulfate reduction and sulfide oxidation in hypoxic sediment in a eutrophic estuary.

Bottom hypoxia and consequential hydrogen sulfide (H2S) release from sediment in eutrophic estuaries is a major global environmental issue. We investigated dissolved oxygen, pH and H2S concentration profiles with microsensors and by sectioning sediment cores followed by colorimetric analysis. The results of these analyses were then compared with the physicochemical properties of the bottom water and sediment samples to determine their relationships with H2S production in sediment. High organic matter and fine particle composition of the sediment reduced the oxidation-reduction potential, stimulating H2S production. Use of a microsensor enabled measurement of H2S concentration profiles with submillimetre resolution, whereas the conventional sediment-sectioning method gave H2S measurements with a spatial resolution of 10 mm. Furthermore, microsensor measurements revealed H2S consumption occurring at the sediment surface in both the microbial mat and the sediment anoxic layer, which were not observed with sectioning. This H2S consumption prevented H2S release into the overlying water. However, the microsensor measurements had the potential to underestimate H2S concentrations. We propose that a combination of several techniques to measure microbial activity and determine its relationships with physicochemical properties of the sediment is essential to understanding the sulfur cycle under hypoxic conditions in eutrophic sediments.

Isotropic plasticity of ß-type Ti-29Nb-13Ta-4.6Zr alloy single crystals for the development of single crystalline ß-Ti implants.

ß-type Ti-29Nb-13Ta-4.6Zr alloy is a promising novel material for biomedical applications. We have proposed a 'single crystalline ß-Ti implant' as new hard tissue replacements for suppressing the stress shielding by achieving a drastic reduction in the Young's modulus. To develop this, the orientation dependence of the plastic deformation behavior of the Ti-29Nb-13Ta-4.6Zr single crystal was first clarified. Dislocation slip with a Burgers vector parallel to <111> was the predominant deformation mode in the wide loading orientation. The orientation dependence of the yield stress due to <111> dislocations was small, in contrast to other ß-Ti alloys. In addition, {332} twin was found to be operative at the loading orientation around [001]. The asymmetric features of the {332} twin formation depending on the loading orientation could be roughly anticipated by their Schmid factors. However, the critical resolved shear stress for the {332} twins appeared to show orientation dependence. The simultaneous operation of <111> slip and {332} twin were found to be the origin of the good mechanical properties with excellent strength and ductility. It was clarified that the Ti-29Nb-13Ta-4.6Zr alloy single crystal shows the "plastically almost-isotropic and elastically highly-anisotropic" nature, that is desirable for the development of 'single crystalline ß-Ti implant'.

Temporal trends for inflow of perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) to Tokyo Bay, Japan, estimated by a receptor-oriented approach.

We estimated inflow rates of perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) to Tokyo Bay, Japan, between February 2004 and February 2011 by a receptor-oriented approach based on quarterly samplings of the bay water. Temporal trends in these inflow rates are an important basis for evaluating changes in PFOS and PFOA emissions in the Tokyo Bay catchment basin. A mixing model estimated the average concentrations of these compounds in the freshwater inflow to the bay, which were then multiplied by estimated freshwater inflow rates to obtain the inflow rates of these compounds. The receptor-oriented approach enabled us to comprehensively cover inflow to the bay, including inflow via direct discharge to the bay. On a logarithmic basis, the rate of inflow for PFOS decreased gradually, particularly after 2006, whereas that for PFOA exhibited a marked stepwise decrease from 2006 to 2007. The rate of inflow for PFOS decreased from 730kg/y during 2004-2006 to 160kg/y in 2010, whereas that for PFOA decreased from 2000kg/y during 2004-2006 to 290kg/y in 2010. These reductions probably reflected reductions in the use and emission of these compounds and their precursors in the Tokyo Bay catchment basin. Our estimated per-person inflow rates (i.e., inflow rates divided by the estimated population in the basin) for PFOS were generally comparable to previously reported per-person waterborne emission rates in Japan and other countries, whereas those for PFOA were generally higher than previously reported per-person waterborne emission rates. A comparison with previous estimates of household emission rates of these compounds suggested that our inflow estimates included a considerable contribution from point industrial sources.

Transfer kinetics of perfluorooctane sulfonate from water and sediment to a marine benthic fish, the marbled flounder (Pseudopleuronectes yokohamae).

The authors investigated the kinetics of transfer of perfluorooctane sulfonate (PFOS) from water, suspended sediment, and bottom sediment to a marine benthic fish, the marbled flounder (Pseudopleuronectes yokohamae). Fish were exposed in 3 treatments to PFOS in combinations of these exposure media for 28 d and then depurated for 84 d. A major part (37-66%) of PFOS in the fish was in the carcass (i.e., whole body minus muscle and internal organs). Three first-order-kinetic models that differed in exposure media, that is, 1) sum of dissolved and particulate phases and sediment; 2) dissolved phase, particulate phase, and sediment; and 3) dissolved phase only, were fitted to the data assuming common rate constants among the treatments. The uptake efficiency of dissolved PFOS at the respiratory surfaces was estimated to be 3.2% that of oxygen, and the half-life of PFOS in the whole body to be 29 d to 31 d. The better fit of models 1 and 2 and the values of the estimated uptake rate constants suggested that the PFOS in suspended and bottom sediments, in addition to that dissolved in water, contributed to the observed body burden of the fish. Based on an evaluation of several possible contributing factors to the uptake of PFOS from suspended and bottom sediments, the authors propose that further investigation is necessary regarding the mechanisms responsible for the uptake.

Temporal changes in carbon and nitrogen stable isotope ratios of macrozoobenthos on an artificial tidal flat facing a hypertrophic canal, inner Tokyo Bay.

Temporal changes in benthic food web structure were analyzed in an artificial tidal flat in inner Tokyo Bay, Japan, using carbon and nitrogen stable isotope ratios (δ(13)C and δ(15)N). Microphytobenthos were the most important food sources of macrozoobenthos, due to high microphytobenthic biomass on the tidal flat, while phytoplankton in canal water (canal POMPP), terrestrial materials from urban surface runoff (canal POMTM), and marsh plants were less important. Dietary contribution of microphytobenthos was highest in April to June, while decreased towards December owing to the supply of canal POMPP and canal POMTM following red tides and heavy rainfall events in summer to fall. Temporal changes in δ(15)N (Δδ(15)N) of consumer corresponded well to the (15)N-enrichment in canal POMPP in summer. A meta-analysis showed that the consumer-Δδ(15)N was considerably larger in inner Tokyo Bay than those in other estuaries, which may be a specific characteristic of benthic food web in highly urbanized estuaries.

Spatial, phase, and temporal distributions of perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) in Tokyo Bay, Japan.

The spatial distribution, partitioning, and time trends of perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) were investigated in the water column and bottom sediment of Tokyo Bay, Japan, during 2004-2006. A total of 480 water and 60 sediment samples obtained by regular 20-station samplings ascertained the three-dimensional distributions of these compounds and changes in the seawater structure in the whole bay. The median of volume-based average water-borne concentrations of PFOS and PFOA was 3.7 and 12 ng/L, respectively. The median concentrations in sediment were 0.61 (PFOS) and 0.20 (PFOA) ng/g-dry. Vertical mixing of the water column probably affected the vertical distribution of these compounds. The negative correlations between PFOS and PFOA concentrations and water salinity and the horizontal distributions of their concentrations suggested that freshwater inputs into the bay were the source of these compounds. A mixing model estimated the average PFOS concentration in the freshwater inputs to be 29 ng/L. The common logarithm of the partition coefficients between the dissolved and suspended-particle-sorbed phases varied among samples, with the average of 4.2 (PFOS) and 3.5 (PFOA). Our analyses indicated no apparent time trends in the concentrations of these compounds during 2004-2006 in either the freshwater input or the bay.

Diversity and similarity of microbial communities in petroleum crude oils produced in Asia.

To understand microbial communities in petroleum crude oils, we precipitated DNA using high concentrations of 2,2,4-trimethylpentane (isooctane) and purified. Samples of DNA from five crude oils, (Middle East, 3; China, 1; and Japan, 1) were characterized based upon their 16S rRNA gene sequences after PCR amplification and the construction of clone libraries. We detected 48 eubacterial species, one cyanobacterium, and one archaeon in total. The microbial constituents were diverse in the DNA samples. Most of the bacteria affiliated with the sequences of the three oils from the Middle East comprised similar mesophilic species. Acinetobacter, Propionibacterium, Sphingobium and a Bacillales were common. In contrast, the bacterial communities in Japanese and Chinese samples were unique. Thermophilic Petrotoga-like bacteria (11%) and several anaerobic-thermophilic Clostridia- and Synergistetes-like bacteria (20%) were detected in the Chinese sample. Different thermophiles (12%) and Clostridia (2%) were detected in the Japanese sample.

Abundance and population structure of ammonia-oxidizing bacteria that inhabit canal sediments receiving effluents from municipal wastewater treatment plants.

A polyphasic, culture-independent study was conducted to investigate the abundance and population structure of ammonia-oxidizing bacteria (AOB) in canal sediments receiving wastewater discharge. The abundance of AOB ranged from 0.2 to 1.9% and 1.6 to 5.7% of the total bacterial fraction by real-time PCR and immunofluorescence staining, respectively. Clone analysis and restriction endonuclease analysis revealed that the AOB communities influenced by the wastewater discharge were dominated by Nitrosomonas, were similar to each other, and were less diverse than the communities outside of the immediate discharge zone.

Characterization and quantification of ammonia-oxidizing bacteria in eutrophic coastal marine sediments using polyphasic molecular approaches and immunofluorescence staining.

Tokyo Bay, a eutrophic bay in Japan, receives nutrients from wastewater plants and other urban diffuse sources via river input. A transect was conducted along a line from the Arakawa River into Tokyo Bay to investigate the ecological relationship between the river outflow and the distribution, abundance and population structure of ammonia-oxidizing bacteria (AOB). Five surficial marine sediments were collected and analysed with polyphasic approaches. Heterogeneity and genetic diversity of beta-AOB populations were examined using restriction fragment length polymorphism (RFLP) analysis of 16S rRNA and amoA genes. A shift of the microbial community was detected in samples along the transect. Both 16S rRNA and amoA genes generated polymorphisms in the restriction profiles that were distinguishable at each sampling site. Two 16S rRNA gene libraries were constructed using the reverse transcription polymerase chain reaction (RT-PCR) method to determine the major ammonia oxidizers maintaining high cellular rRNA content. Two major groups were observed in the Nitrosomonas lineage; no Nitrosospira were detected. The effort to isolate novel AOB was successful; the isolate dominated in the gene libraries. For quantitative analysis, a real-time PCR assay targeting the 16S rRNA gene was developed. The population sizes of beta-AOB ranged from 1.6 x 10(7) to 3.0 x 10(8) cells g(-1) in dry sediments, which corresponded to 0.1-1.1% of the total bacterial population. An immunofluorescence staining using anti-hydroxylamine oxidoreductase (HAO) antibody was also tested to obtain complementary data. The population sizes of ammonia oxidizers ranged between 2.4 x 10(8) and 1.2 x 10(9) cells g(-1) of dry sediments, which corresponded to 1.2-4.3% of the total bacterial fraction. Ammonia-oxidizing bacteria cell numbers deduced by the two methods were correlated (R = 0.79, P < 0.01). In both methods, the number of AOB increased with the distance from the river mouth; ammonia-oxidizing bacteria were most numerous at B30, where the ammonium concentration in the porewater was markedly lower and the nitrite concentration was slightly higher than nearby sites. These results reveal spatial distribution and shifts in the population structure of AOB corresponding to nutrients and organic inputs from the river run-off and phytoplankton bloom.

Electrostatic interaction between redox cofactors in photosynthetic reaction centers.

Intramolecular electron transfer within proteins is an essential process in bioenergetics. Redox cofactors are embedded in proteins, and this matrix strongly influences their redox potential. Several cofactors are usually found in these complexes, and they are structurally organized in a chain with distances between the electron donor and acceptor short enough to allow rapid electron tunneling. Among the different interactions that contribute to the determination of the redox potential of these cofactors, electrostatic interactions are important but restive to direct experimental characterization. The influence of interaction between cofactors is evidenced here experimentally by means of redox titrations and time-resolved spectroscopy in a chimeric bacterial reaction center (Maki, H., Matsuura, K., Shimada, K., and Nagashima, K. V. P. (2003) J. Biol. Chem. 278, 3921-3928) composed of the core subunits of Rubrivivax gelatinosus and the tetraheme cytochrome of Blastochloris viridis. The absorption spectra and orientations of the various cofactors of this chimeric reaction center are similar to those found in their respective native protein, indicating that their local environment is conserved. However, the redox potentials of both the primary electron donor and its closest heme are changed. The redox potential of the primary electron donor is downshifted in the chimeric reaction center when compared with the wild type, whereas, conversely, that of its closet heme is upshifted. We propose a model in which these reciprocal shifts in the midpoint potentials of two electron transfer partners are explained by an electrostatic interaction between them.

Effect of dietary medium- and long-chain triacylglycerols (MLCT) on accumulation of body fat in healthy humans.

We investigated whether a structured medium- and long-chain triacylglycerols (MLCT) diet could decrease accumulation of body fat in healthy humans. The study was conducted under a double-blind randomized design. Ninety-three subjects participated in this study. However, 10 subjects could not consume the specified meal, and one subject wished to opt out. Consequently, the study included 82 subjects. The experimental subjects consumed the test bread, which was made with 14 g of MLCT containing 1.7 g MCFA, daily at breakfast during the study period of 12 weeks, and the control subjects consumed bread made with long-chain triacylglycerols (LCT). All subjects consumed the same standard packaged meals. Body composition parameters were body weight, total body fat and abdominal fat, and blood analyses included serum cholesterol, triacylglycerols and phospholipids. Significant decreases of body weight, the amount of body fat, subcutaneous and visceral fat were noted in the MLCT group as compared with those of the LCT group for 12 weeks (P<0.05). Furthermore, a significant decrease in serum total cholesterol was noted in the MLCT group as compared with that of the LCT group at 8 weeks (P<0.05). However, other serum parameters were not different between the MLCT and LCT groups. The results suggest that the daily intake of MLCT diet could result in a reduction in body weight and in accumulation of body fat, and, moreover, it could reduce serum total cholesterol.

Crude oil bioremediation field experiment in the Sea of Japan.

Experimental bioremediation of crude oil was conducted for approximately 3 months in the intertidal zone of the Sea of Japan, Hyogo Prefecture. Artificial mixtures of weathered Arabian light crude oil and sand taken from the experimental site were wrapped in polyester net envelopes. The envelopes were placed in drum-shaped acrylic vessels with perforated sides to facilitate seawater exchange. The vessels were laid in the intertidal area. Slow release nitrogen and phosphorus synthetic fertilizer granules were added to the oil-sand mixtures in three different amounts. Some oil-sand mixtures were unfertilized controls. The oil-sand mixtures were periodically sampled and changes in the composition of the residual oils were monitored. Oil samples were subjected to gas chromatography coupled with mass spectrometry for analysis of some representative semi-volatile aliphatic and aromatic compounds. All values for each analyte were normalized against that of hopane to evaluate the extent of oil biodegradation. Significant increases in the concentrations of both nitrogen and phosphorus were found in the fertilized sections in accordance with the amounts of added fertilizers. Although significant natural attenuation of oil was observed in the unfertilized sections, fertilization stimulated the degradation rate of the oil in the early stage of the experimental term. The extent of the oil biodegradation increased as the amount of added fertilizer increased. However, the final degradation efficiencies for each oil component in the fertilized sections were not significantly different from those in the unfertilized sections, and the degradation of each oil component had almost ceased after 6 weeks. We conclude that excessive amounts of macronutrients are required to accelerate oil biodegradation and that fertilization is only effective in the early stages.

Monitoring of alkane-degrading bacteria in a sea-water microcosm during crude oil degradation by polymerase chain reaction based on alkane-catabolic genes.

Behaviour of microbial populations responsible for degrading n-alkanes, a major component of crude oil, was monitored during crude oil degradation in a sea-water microcosm by both traditional colony culturing and molecular techniques. A DNA extraction method applicable to crude oil-amended sea-water samples was developed to obtain DNA applicable to most probable number (MPN) polymerase chain reaction (PCR). The population of alkane-degrading bacteria responsible for degradation of n-alkanes in a crude oil-amended microcosm altered, so that shorter alkanes were degraded first by alkane-degrading bacteria possessing alkane hydroxylase genes from group I (Kohno et al., 2002, Microb Environ 17: 114-121) and longer ones afterwards by those possessing alkane hydroxylase genes from group II. Thus, the degradation mechanism of the n-alkanes can be clarified during crude oil degradation. Application of the method of detecting different types of alkane-catabolic genes, as shown in the present study, enabled bacterial groups preferring alkanes of either shorter or longer chain lengths to be enumerated selectively.

Effect of medium-chain triglycerides on the postprandial triglyceride concentration in healthy men.

This study compared the serum lipid concentrations after a single dose of medium-chain triglycerides (MCT) or long-chain triglycerides (LCT) between individuals grouped according to the body mass index (BMI). Twenty-five males participated as volunteers, the test diet containing 10 g of MCT or LCT. Blood samples were collected up to 6 h after the intake of a test diets. The LCT diet resulted in significantly greater increases in areas under the curves (AUCs) for serum and chylomicron triglyceride in the BMI > or = 23 kg/m2 group than those in the BMI < 23 kg/m2 group. The magnitude of response after intake of the MCT diet by the BMI > or = 23 kg/m2 group was significantly lower than that after the LCT diet. These results suggest that, in subjects with BMI > or = 23 kg/m2, the intake of MCT is preferable to that of LCT for maintaining postprandial triglyceride at a low concentration.

Chimeric photosynthetic reaction center complex of purple bacteria composed of the core subunits of Rubrivivax gelatinosus and the cytochrome subunit of Blastochloris viridis.

A gene coding for the photosynthetic reaction center-bound cytochrome subunit, pufC, of Blastochloris viridis, which belongs to the alpha-purple bacteria, was introduced into Rubrivivax gelatinosus, which belongs to the beta-purple bacteria. The cytochrome subunit of B. viridis was synthesized in the R. gelatinosus cells, in which the native pufC gene was knocked out, and formed a chimeric reaction center (RC) complex together with other subunits of R. gelatinosus. The transformant was able to grow photosynthetically. Rapid photo-oxidization of the hemes in the cytochrome subunit was observed in the membrane of the transformant. The soluble electron carrier, cytochrome c(2), isolated from B. viridis was a good electron donor to the chimeric RC. The redox midpoint potentials and the redox difference spectra of four hemes in the cytochrome subunit of the chimeric RC were almost identical with those in the B. viridis RC. The cytochrome subunit of B. viridis seems to retain its structure and function in the R. gelatinosus cell. The chimeric RC and its mutagenesis system should be useful for further studies about the cytochrome subunit of B. viridis.

Effects of margarine containing medium-chain triacylglycerols on body fat reduction in humans.

We found previously that the ingestion of margarine containing medium-chain triacylglycerols (MCT) resulted in a significant increase in postprandial thermogenesis when compared with long-chain triacylglycerols (LCT). Diets that included margarine containing MCT and LCT were compared for 12 weeks in 73 subjects to investigate the effects on body weight, body fat, areas of subcutaneous and visceral fat, serum total cholesterols, triglycerides, lipoproteins, plasma glucose, serum insulin, total ketone bodies, and the activities of aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyltranspeptidase. We conducted a double-blind, controlled study and used blended rapeseed oil and soybean oil (LCT) as a comparison. Two groups ingested 2100-2400 kcal/day of energy, 65-73 g/day of total fat, and 14 g/day of test margarine (5 g/day of MCT or LCT). The subjects on the MCT diet demonstrated significant decreases in body fat weight (- 3.8 +/- 2.4 kg vs - 2.4 +/- 1.7 kg; MCT vs LCT, mean +/- SD), subcutaneous fat (- 38.2 +/- 29.9 cm(2) vs - 22.6 +/- 19.3 cm(2)), and visceral fat (- 12.2 +/- 11.2 cm(2) vs - 1.6 +/- 12.8 cm(2)) after 12 weeks. There were no clinical differences in measured blood parameters. We suggest that the postprandial increase in thermogenesis and control of postprandial triglyceride levels may explain these results.

Comparison of diet-induced thermogenesis of foods containing medium- versus long-chain triacylglycerols.

The purpose of this study was to investigate the effect of 5-10 g of medium-chain triacylglycerols (MCT) on diet-induced thermogenesis in healthy humans. The study compared diet-induced thermogenesis after ingestion of test foods containing MCT and long-chain triacylglycerols (LCT), using a double-blind, crossover design. Eight male and eight female subjects participated in study 1 and study 2, respectively. In both studies, the LCT was a blend of rapeseed oil and soybean oil. In study 1, the liquid meals contained 10 g MCT (10M), a mixture of 5 g MCT and 5 g LCT (5M5L), and 10 g LCT (10L). In study 2, the subjects were given a meal (sandwich and clear soup) with the mayonnaise or margarine containing 5 g of MCT or LCT. Postprandial energy expenditure was measured by indirect calorimetry before and during the 6 h after ingestion of the test meals. Diet-induced thermogenesis was significantly greater after 5M5L and 10M Ingestion as compared to 10L ingestion. Ingestion of the mayonnaise or margarine containing 5 g MCT caused significantly larger diet-induced thermogenesis as compared to that of LCT. These results suggest that, in healthy humans, the intake of 5-10 g of MCT causes larger diet-induced thermogenesis than that of LCT, irrespective of the form of meal containing the MCT.