Degradation of outer membrane proteins of <Emphasis Type="Italic">Synechococcus</Emphasis> sp. <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis>

To examine the possibility that outer membrane proteins (OMP) of Synechococcus sp. remain in seawater, we investigated the stability of OMPs in vitro and in situ. Some fractions prepared from Synechococcus sp. CSIRO-94 were treated with trypsin and proteinase K. Four tightly bound OMPs were separated from Synechococcus. We designated the two major OMPs of 52 kDa and 48 kDa as Omp52Sy and Omp48Sy, respectively. Degradation of the OMP in natural seawater was monitored in microcosms to which intact Synechococcus cells and outer membrane (OM) were added. Omp52Sy and Omp48Sy were the most stable against trypsin and proteinase K among the OMPs when they were embedded in the OM. However, in the microcosm experiment using intact cells, Omp52Sy and Omp48Sy were detected in the particulate fraction only during the first 4 days, after which they could not longer be detected. Omp52Sy and Omp48Sy were the most stable proteins among the Synechococcus OMPs in vitro, but they might be degraded in situ. This indicates that stability of Synechococcus porin differs depending on complex formation with other membrane molecules, which might cause different preservation of microbial membrane proteins in the dissolved protein pool in the ocean. This study suggests that Gram negative bacterial OM with thin peptidoglycan forms a lipid bilayer that proptects OMP, but Synechococcus OM with thick peptidoglycan cannot form a lipid bilayer. The incomplete bilayer might not be able to protect from protease attack in the natural environment.     

Temporal variability and characterization of physicochemical properties in the neritic area of Sagami Bay,Japan

Seasonal and interannual variations in physicochemical properties were investigated in the neritic area of Sagami Bay, Kanagawa, Japan, from December 2000 to December 2005. Physicochemical properties (i.e. temperature, salinity, density, dissolved oxygen and dissolved inorganic nutrient concentration) revealed clear seasonal variations, which were similar to each other during all 5 years. Temperature, salinity and dissolved inorganic nutrients showed rapid, drastic variations within a few days and/or weeks. These variations are related to sea levels, principally due to the shifting effects of the Kuroshio Current axis: they were strongly affected by the Kuroshio Water and other waters, when sea level difference was greater than ca. 35 cm and lower than ca. 15 cm, respectively. Temperature difference (DF _T ) increased with sea level difference, and the difference of salinity and dissolved inorganic nutrients (NH₄ ⁺-N, NO₃ ⁻+NO₂ ⁻-N, NH₄ ⁺+NO₃ ⁻+NO₂ ⁻-N, PO₄ ³⁻-P and SiO₂-Si) increased and decreased with DF _T , respectively. All these correlations are significant. Total dissolved inorganic nitrogen (N), phosphate (P) and silicate (Si) revealed seasonal variations in the ranges of 0.57–16.08, 0.0070–0.91 and 0.22–46.38 μM, respectively. From the regression equations between these elements allowed the following relation to be obtained; Si:N:P = 14.8:13.4:1. Dissolved inorganic nutrients were characterized by Si and/or P deficiency, especially in the upper layer (0–20 m depth) during summer. Single and/or combined elements are discussed on the basis of potential and stoichiometric nutrient limitations, which could restrict phytoplankton (diatom) growth as a limiting factor.     

Vertical Distributions of Larvae of the Clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> and the Striped Horse Mussel <Emphasis Type="Italic">Musculista senhousia</Emphasis> in Eastern Ariake Bay,Southern Japan

We have examined the vertical distributions of planktonic bivalve larvae, particularly the clam Ruditapes philippinarum and the mussel Musculista senhousia which are common and abundant on tidal flats of eastern Ariake Bay, southern Japan. Submersible pumping gear was used to take samples at 2 stations every 2 hours during the spring tide over a whole tidal cycle and/or through daytime and nighttime. Water samples were pumped up from 3 to 5 depths from the surface to sea bottom. Regardless of tidal cycles, D-shaped larvae were concentrated near the surface, while umbo larvae were found at the surface to intermediate depths. On average, these larval densities were significantly higher at the station close to the shore than the offshore station, with no significant difference between daytime and nighttime and between flood and ebb tides at each station. The velocity and direction of water movement at both stations revealed remarkable difference between the surface and bottom waters. The larvae at the surface may quickly disperse and be transported elsewhere, while those in water close to the sea bottom may tend to be retained.     

Excretion and respiration rates of the scyphomedusa <Emphasis Type="Italic">Aurelia aurita</Emphasis> from the Inland Sea of Japan

We measured the ammonium excretion, phosphate excretion and respiration rates of the scyphomedusa Aurelia aurita from Ondo Strait, in the central part of the Inland Sea of Japan, at 28 and 20°C. The rates measured at 28°C were converted to those at 20°C using the Q₁₀ values, i.e. 1.56, 1.57 and 2.80, for ammonium excretion, phosphate excretion and respiration rates, respectively. The composite relationships between metabolic rates and wet weight of a medusa (WW, g, range 11–1330 g) at 20°C were expressed by the following allometric equations. For ammonium excretion rate (N, μmoles N medusa⁻¹d⁻¹): N = 0.497WW ^1.09, phosphate excretion rate (P, μmoles P medusa⁻¹d⁻¹): P = 0.453WW ^0.84, and respiration rate (R, μmoles O₂ medusa⁻¹d⁻¹): R = 96.9WW ^1.06. Mean O:N ratios (i.e. atomic ratios of 16.9 and 11.0 at 28 and 20°C, respectively) indicated that the metabolism of A. aurita medusae was protein-dominated. These metabolic parameters enabled us to estimate the nitrogen and phosphorus regeneration rates of an A. aurita medusa population typical of early summer in the Ondo Strait (means of water temperature, medusa individual weight and population biomass: 20°C, 200 g WW and 50.8 g WW m⁻³, respectively). Regenerated nitrogen and phosphorus were equivalent to 10.0 and 21.6% of phytoplankton uptake rates, respectively, nearly twice that estimated for mesozooplankton, demonstrating that A. aurita medusae are key components of the plankton community, influencing the trophic and nutrient dynamics in the Ondo Strait during early summer.     

Crown foliage dynamics of mangrove <Emphasis Type="Italic">Kandelia obovata</Emphasis> in Manko Wetland,Okinawa Island,Japan

The crown foliage dynamics of Kandelia obovata (S., L.) Yong were investigated, including leaf recruitment, survival and leaf growth. Newly flushed leaves occurred successively throughout the year, with a maximum in July and a minimum in January. The highest leaf death was in August, whereas it was the lowest in January. The relative mortality rate of leaves seemed to depend on the season when new leaves flushed. The growth pattern of leaves varied among seasons. The maximum leaf area was significantly larger in winter than in autumn, but showed no significant difference between the winter and the other seasons. The half-expansion period and the intrinsic rate of increase were respectively longer and lower in winter than in the other seasons. Therefore, leaves flushed in summer grew faster in their initial stage and attained their maximum leaf area sooner than those flushed in winter. This most likely results from the difference in temperature between summer and winter. The crown leaf area was almost stable throughout the year, as if homeostatic control is likely to function. Stipule litterfall decreased significantly with increasing flower and propagule litterfalls, indicating that a high production of flowers and propagules results in low leaf recruitment. This may be because flower production and propagule growth impose a severe burden on K. obovata.     

Genetic diversity and population structure of three dominant myctophid fishes (<Emphasis Type="Italic">Diaphus theta,Stenobrachius leucopsarus</Emphasis>, and <Emphasis Type="Italic">S. nannochir</Emphasis>) in the North Pacific Ocean

We compared the genetic diversity of three dominant myctophid fishes in the North Pacific Ocean that have different diel vertical migration patterns on the basis of the nucleotide sequences of the mitochondrial gene for cytochrome b. No genetic structure was detected for each of these three species. The genetic diversity progressively increased for Diaphus theta, a diel migrant species showing clear diel vertical migration; Stenobrachius leucopsarus, a semi-diel migrant, in which only part of the population migrates vertically; S. nannochir, a non-diel migrant. All three species were suggested to have experienced a recent, sudden population expansion. Interspecific differences in genetic diversity might be attributable to differences in the degree of population size reduction during the glacial periods; this degree in turn corresponds to the energy demand of the fishes.     

Production of <Emphasis Type="Italic">Acartia steueri</Emphasis> (Copepoda: Calanoida) in Ilkwang Bay,Southeastern Coast of Korea

Production of the marine calanoid copepod Acartia steueri was measured from 2 October 1991 to 8 October 1992 at a station in Ilkwang Bay, on the southeastern coast of Korea. Phytoplankton standing stock ranged over 1.0 to 9.3 mg chl.a m⁻³, and annual primary productivity (by the C-14 method) at three stations was estimated at 200 gC m⁻² yr⁻¹. Acartia steueri (nauplii + copepodids + adults) were present in the plankton throughout the year, with seasonal variation in abundance. Biomass of A. steueri, excluding the NI stage, was 0.01–4.55 mgC m⁻³ (mean: 0.68 mgC m⁻³) with peaks in November, February, May and July-early August, and relatively low biomass in September– January. Instantaneous growth rates of the nauplius stages were higher than the copepodid stages. Annual production of A. steueri was 25.1 mgC m⁻³ yr⁻¹ (or 166 mgC m⁻² yr⁻¹), showing peaks in November, May and July–August with a small peak in February, and low production in December–April and September–October. There were no significant relationships between the daily production rate of A. steueri and temperature or chlorophyll a concentration, indicating that unknown other factors might be related to the variation of the production rate.     

Numerical experiment on <Emphasis Type="Italic">Kyucho</Emphasis> around the Tango Peninsula induced by Typhoon 0406

A numerical experiment using a three dimensional level model was performed to clarify the mechanism generating a strong coastal current, Kyucho, induced by the passage of Typhoon 0406 around the tip of the Tango Peninsula, Japan in June 2004. Wind stress accompanied by Typhoon 0406 was applied to the model ocean with realistic bottom topography and stratification condition. The model well reproduced the characteristics of Kyucho observed by Kumaki et al. (2005), i.e., the strong alongshore current with maximum velocity of 53 cm s⁻¹ and its propagation along the peninsula with propagation speed of about 0.6 m s⁻¹ one half-day after the typhoon’s passage. Coastal-trapped waves (CTW) accompanied by downwelling were induced along the northwest coast of the peninsula by the alongshore wind stress. The energy density flux due to the CTW flowed eastward along the coast, and indicated scattering of the CTW around the eastern coast of the peninsula. In addition, significant near-inertial internal gravity waves were also caused in the offshore region from the west of the Noto Peninsula to the north of the Tango Peninsula by the typhoon’s passage. The energy flux density of the near-inertial fluctuations flowed southward off the Fukui coast, and part of the energy flux was trapped on the tip of the Tango Peninsula, flowing with the coast on its right. It was found that the strong current, Kyucho, at the northeastern tip of the Tango Peninsula was generated by superposition of the near-inertial internal gravity waves and subinertial CTW.     

Distribution of ephyrae and polyps of jellyfish <Emphasis Type="Italic">Aurelia</Emphasis><Emphasis Type="Italic">aurita</Emphasis> (Linnaeus 1758) <Emphasis Type="Italic">sensu lato</Emphasis> in Mikawa Bay,Japan

We surveyed the distribution of colonies of polyps of Aurelia aurita sensu lato (s.l.) in Mikawa Bay, Japan. First, we surveyed the distribution of ephyrae of A. aurita s.l. at 75 stations encompassing the whole of Mikawa Bay in early 2008. A total of 37 ephyrae were sampled mostly from fishing ports. Ephyrae were most abundant around the islands located near the mouth of the bay, and decreased from the western part to the eastern part of Mikawa Bay. Next, we selected five fishing ports in Mikawa Bay where ephyrae occurred and surveyed the underside of floating piers and underwater overhangs of wharfs. We found dense colonies of polyps of A. aurita s.l. under nearly all of the floating piers at the two islands located near the mouth of the bay. Fitting a logistic regression model to the dataset showed that the percentage coverage of Aurelia polyps was significantly greater at the two islands compared with the other locations. In addition, the coverage of Aurelia polyps was greater when the coverage of other fouling organisms was in the range of 65–90%, and the coverage of Aurelia polyps was lower on floating piers with a vinyl surface and on concrete wharfs. The combined distribution of polyp colonies of A. aurita s.l. in Ise Bay and Mikawa Bay suggested that A. aurita s.l. in the two bays probably forms a single population and shoals of medusae mainly originate from protected harbors along the mouth-part of the bays.     

Molecular ecological study of <Emphasis Type="Italic">Siganus spinus</Emphasis> and <Emphasis Type="Italic">S. guttatus</Emphasis> from Okinawan waters based on mitochondrial DNA control region sequences

The mitochondrial DNA (mtDNA) control region was sequenced to examine the genetic variability and gene flow of juvenile Siganus spinus and S. guttatus. In total, 461 nucleotide sequences were obtained from 69 specimens of juvenile S. spinus from Okinawa Island and Ishigaki Island, in which 17 variable sites and 22 haplotypes were identified. Haplotype diversity (h) was high (0.9244 in Okinawa and 0.8984 in Ishigaki), whereas nucleotide diversity (π) was low (0.0063 in Okinawa and 0.0059 in Ishigaki). The two populations were not genetically distinct. Siganus guttatus, which do not form large schools at recruitment, in contrast S. spinus, were also analyzed by studying 152 individuals collected off Okinawa Island, Miyako Island, and Ishigaki Island. Of 476 nucleotide sequences, 50 were variable, and 42 haplotypes were identified. Genetic variability values were high (h = 0.8766 and π = 0.0151 in Okinawa; h = 0.9640 and π = 0.0192 in Miyako; h = 0.9161 and π = 0.0199 in Ishigaki). The Okinawa population was genetically isolated from the Miyako and Ishigaki populations. As a result, genetic diversity was high for each of these siganid populations despite their being target species for fisheries; however, the degree of inter-population gene flow was higher for S. spinus than S. guttatus, suggesting that these species exhibit different dispersal strategies.     

Age and Growth of <Emphasis Type="Italic">Sillago aeolus</Emphasis> in Okinawa Island,Japan

In a study of the age and growth of Sillago aeolus, a total of 414 fish were collected from December 2000 to March 2002, around Okinawa Island. Ages of 403 specimens were determined by otoliths. Monthly changes in the percentage of otoliths with an opaque zone in the outer margin indicated that an opaque ring formed once a year, occurring between February and May. The rings can therefore be defined as annual rings. Ages of females were estimated to be 0+ to 4+, while those of males were 0+ to 2+. The von Bertalanffy growth curves were well fitted to age-standard length data of males and females, as follows: Male: L_t = 209.6 [1 − exp{−0.70 (t + 0.58)}]. Female: L_t = 297.7 [1 − exp{−0.42 (t + 0.61)}].     

Characterization of proteins in the muscle of<Emphasis Type="Italic">limanda yokohamae</Emphasis> from the masan bay,Korea

Increasing industrial development in the Masan Bay area of Korea over the past decades increased the risk for the survival of marine organisms in the bay area by the deterioration of the water quality. Since living organisms have the ability to adapt contamination-associated stimuli by the alteration of gene expression, changes in proteins can be used as an important criterion for assessing the levels of environmental conditions. In this study, therefore, alterations of the expression of proteins in the muscle ofLimanda yokohamae from Dukdong and Dotsum in the bay area were surveyed and characterized as compared with Haegumgang, which served as a control site. The results demonstrated that the twenty spots detected from Dukdong and Dotsum were similar to each other. Fifteen proteins were found to be predicted or undefined proteins, while five proteins were identified as heavy polypeptide 11 of myosin, apolipoprotein A-I, fibroblast growth factor 17b precursor, G protein-coupled receptor kinase 1 b and bonnie and clyde. These data suggest that local fish in the bay area have dysfunction in muscle physiology including contraction, lipid metabolism, proliferation and differentiation and nervous system.     

Turbulent structure of the subsurface layer of the sea according to the data of the <Emphasis Type="Italic">Sigma-1</Emphasis> measuring complex

We present the results of experimental investigations of the characteristics of turbulence in the layer of wave-induced mixing. The data on the fluctuations of velocity, temperature, and conductivity are obtained with the help of a Sigma-1 measuring complex. The computed values of the dissipation rate of turbulent energy are compared with different models proposed for the subsurface layer. It is shown that the available models fail to guarantee satisfactory agreement of the numerical results with the experimental data for the layer of active wave action and, in particular, in the presence of swell. This leads us to the conclusion concerning the necessity of parametrization and assimilation of more complete data on the state of the sea surface, the structure of currents, and the surface layer of the atmosphere in the models. __________ Translated from Morskoi Gidrofizicheskii Zhurnal, No. 2, pp. 15–28, March–April, 2007.     

The transition zone chlorophyll front can trigger <Emphasis Type="Italic">Acanthaster planci</Emphasis> outbreaks in the Pacific Ocean: Historical confirmation

We hypothesize that the North Pacific transition zone chlorophyll front (TZCF) can episodically deliver enhanced phytoplankton levels that are linked to the emergence of adult populations of the coral eating starfish Acanthaster planci. In some years, the seasonally migrating TZCF bathes the northwest Hawaiian Islands with chlorophyll-a rich waters during the winter months that coincide with peak starfish spawning and provide ideal conditions for A. planci larval survival. We found significant relationships between starfish populations in the North Pacific and the southernmost latitude of the TZCF, chlorophyll-a concentrations, sea surface temperature, and Ekman transport indices since 1967. We propose that TZCF-triggered primary outbreaks are followed by secondary outbreaks throughout the region, in accordance with the surface currents and separated by a sequential time lag. Our historical confirmation suggests outbreaks are predictable, which has immediate coral reef conservation and management consequences.     

Stage-V copepodites of Calanus sinicus and Calanus jashnovi (Copepoda: Calanoida) in mesopelagic zone of Sagami Bay as identified with genetic markers, with special reference to their vertical distribution

We applied genetic makers to identify Calanus species occurring in Sagami Bay, Japan, in order to investigate their vertical distribution in the upper 1000 m. First, interspecific genetic distances of three gene loci, mitochondrial small ribosomal RNA (srRNA), nuclear internal transcribed spacers 1 (ITS1) and 2 (ITS2), were estimated from morphologically distinguishable adult females of Calanus sinicus, Calanus jashnovi and Calanus pacificus that were collected from Sagami Bay, the Kuroshio Extension and the Oyashio region, respectively. The highest levels of interspecific genetic distance were observed in srRNA, followed by ITS1 and ITS2. The intraspecific genetic distances within C. sinicus were much lower than the interspecific genetic distances, indicating that DNA sequences in these loci are consistent with the morphological differences. This information was used as a criterion for species identification based on DNA sequence variation, and allowed us to identify the fifth copepodites (CVs) or younger stages of these species. Next, the vertical distribution of Calanus species was investigated in Sagami Bay in May 2006, on the basis of a stratified sampling in the upper 1000 m. By applying the genetic markers, 23 individuals comprising all copepodite stages were allocated into either C. sinicus or C. jashnovi, and the small- and large-sized CVs were identified as C. sinicus and C. jashnovi, respectively. The total abundance of C. sinicus was highest at 0-50 m and decreased with depth. On the contrary, CV individuals of C. sinicus were abundant not only in 0-50 m but also below 200 m with minimum occurrences in 150-200 m depth. C. jashnovi was much less abundant than C. sinicus and comprised of only CIV and CV which occurred in the upper 100 m and deeper than 50 m depths, respectively. The abundance of C. sinicus in the 1000-m water column of Sagami Bay was at a level comparable to that in shelf waters, suggesting the importance of off-shelf individuals in the biological production and organic transport in the respective areas.     

Direct examination of the dietary preference of the copepod<Emphasis Type="Italic">calanus helgolandicus</Emphasis> using the colorimetric approach

The food selectivity of tethered females of the copepodCalanus helgolandicus was examined by using the colorimetric approach. First, feeding behavior of the copepod did not show any differences between the red-color stained with neutral red and non-stained diets using the diatomCoscinodiscus curvatulus. Then, the copepods were fed a mixtures of two diets, the diatomC. curvatulus, stained with neutral red, and the dinoflagellateGymnodinium sanguineum for 14~60 minutes of feeding duration. The foregut colors of females were examined using a stereo-microscope and a video monitor. The foreguts of animals fed with a high density of diatoms in mixed diets showed a dark red color, whereas those fed with a high density of dinoflagellate in mixed diets were a dark yellow. The results suggest that this species of copepod may not selectively feed either one of the diets used for this study. Their feeding activity may be more likely related to the density of available prey in their environment. Therefore, this quick and easy colorimetric approach could provide very useful information, like the pre-screening procedure before designing and conducting the time-consuming and complex feeding experiments to understand the feeding ecology of copepods.     

Intercellular localization of nitrogenase in a non-heterocystous cyanobacterium (cyanophyte), <Emphasis Type="Italic">Trichodesmium</Emphasis> sp. NIBB1067

The nitrogenase activities of cyanobacteria of the genus Trichodesmium are strictly light-dependent, although they do not develop heterocysts. Recently, the development of heterocyst-equivalent cells (diazocytes) was suggested in Trichodesmium spp. However, no cells with a similar appearance to diazocytes could be found in Trichodesmium sp. NIBB1067. An immunocytochemical analysis of nitrogenase in individual cells was performed using polyclonal antibodies generated from DNA fragments of genes encoding the Fe-protein and the α-and the β-subunit of the MoFe-protein of nitrogenase of Trichodesmium sp. NIBB1067. Visualization of the antibody binding was carried out using a horseradish peroxidase-conjugated secondary antibody and a chromogenic substrate, 3,3′-diaminobenzidine, to avoid the masking effects caused by the bright autofluorescence emitted by Trichodesmium cells. Nitrogenase proteins were detected in almost all the cells (always higher than 95% of total cells) grown under nitrogen-limited conditions. These results indicate that Trichodesmium sp. NIBB1067 does not differentiate heterocyst-equivalent cells.