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.     

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.     

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.     

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.     

Reproductive ecology of the dominant dinoflagellate, <Emphasis Type="Italic">Ceratium fusus</Emphasis>, in coastal area of Sagami Bay,Japan

The seasonal abundance of the dominant dinoflagellate, Ceratium fusus, was investigated from January 2000 to December 2003 in a coastal region of Sagami Bay, Japan. The growth of this species was also examined under laboratory conditions. In Sagami Bay, C. fusus increased significantly from April to September, and decreased from November to February, though it was found at all times through out the observation period. C. fusus increased markedly in September 2001 and August 2003 after heavy rainfalls that produced pycnoclines. Rapid growth was observed over a salinity range of 24 to 30, with the highest specific rate of 0.59 d⁻¹ measured under the following conditions: salinity 27, temperature 24°C, photon irradiance 600 μmol m⁻²s⁻¹. The growth rate of C. fusus increased with increasing irradiance from 58 to 216 μmol m⁻²s⁻¹, plateauing between 216 and 796 μmol m⁻²s⁻¹ under all temperature and salinity treatments (except at a temperature of 12°C). Both field and laboratory experiments indicated that C. fusus has the ability to grow under wide ranges of water temperatures (14–28°C), salinities (20–34), and photon irradiance (50–800 μmol m⁻²s⁻¹); it is also able to grow at low nutrient concentrations. This physiological flexibility ensures that populations persist when bloom conditions come to an end.     

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.     

Morphological differences between three groups of <Emphasis Type="Italic">Luciogobius guttatus</Emphasis> on the Korean Peninsula

Three ecological and geographical forms of Luciogobius guttatus from the western and eastern coasts of the Korean Peninsula as well as the coasts of, Jeju Island which is located to the south of the peninsula are described and compared based on morphological characteristics. The environments of these three coastal areas are different, and the morphometrics show a clear pattern of differentiation and reveal the discreteness of the western, eastern and Jeju groups. In addition, a discriminant analysis (DA), a principal component analysis (PCA) of morphometric characteristics, and a t-test for meristic characteristics all show that the three populations of L. guttatus differ from each other. Three divergent lineages of L. guttatus are described with the possibility of niche differentiation between the western, eastern and Jeju coastal habitats. The standard length of eastern populations was greater than that of western and Jeju populations, and those with morphometric characteristics had a longer body. In addition, eastern populations had a higher number of vertebrae. We speculate that the characteristics of eastern populations evolved through the difference in tide level and gravel density, whereas the difference between western and Jeju populations is due to geographical distance.     

Molecular phylogeny of the genus <Emphasis Type="Italic">Chondracanthus</Emphasis> (Rhodophyta), focusing on the resurrection of <Emphasis Type="Italic">C. okamurae</Emphasis> and the description of <Emphasis Type="Italic">C. cincinnus</Emphasis> sp. nov.

Determining the taxonomic status of the red algal genus Chondracanthus based on morphological characters is challenging due to the similarity and high degree of plasticity of the thallus. Since the taxonomic history of several Chondracanthus species remains unclear, we analyzed the plastid rbcL and mitochondrial COI genes of the specimens from Korea and Japan, in combination with morphological observations, to examine their phylogenetic relationships. Our results confirmed the distinction of C. okamurae, which is separated from C. intermedius, and identified a novel species, C. cincinnus sp. nov. Three species (C. okamurae, C. intermedius and C. cincinnus) formed a monophyletic clade with C. tenellus. C. okamurae is distinguished by linear, narrow, cylindrical to compressed, slightly recurved axes, and a high-intertidal to subtidal distribution. It was collected from Korea and Japan, while C. intermedius was identified from Japan only. A new species, Chondracanthus cincinnus sp. nov., is characterized by linear, compressed, strongly recurved axes, and a low-intertidal to subtidal distiribution. Based on the molecular phylogeny using rbcL and COI data, we herein resurrect C. okamurae as a distinct species and identify C. cincinnus as a new species.     

New polymorphic microsatellite markers derived from hemocyte cDNA library of Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> challenged by the protozoan parasite <Emphasis Type="Italic">Perkinsus olseni</Emphasis>

Manila clam Ruditapes philippinarum is one of the most important benthic animals in the coastal north Pacific region, where clam populations have been mixed genetically through trade and aquaculture activities. Accordingly, identification of the genetically different clam populations has become one of the most important issues to manage interbreeding of the local and introduced clam populations. To identify genetically different populations of clam populations, we developed 11 expressed sequence tag (EST)-microsatellite loci (i.e., simple sequence repeat, SSR) from 1,128 clam hemocyte cDNA clones challenged by the protozoan parasite Perkinsus olseni. Genotype analysis using the markers developed in this study demonstrated that clams from a tidal flat on the west coast contained 6 to 19 alleles per locus, and a population from Jeju Island had 4 to 20 alleles per locus. The expected heterozygosity of the 2 clam populations ranged from 0.472 to 0.919 for clams from the west coast, and 0.494 to 0.919 for clams from Jeju Island, respectively. Among the 11 loci discovered in this study, 7 loci significantly deviated from the Hardy-Weinberg equilibrium after Bonferroni correction. The 5 loci developed in this study also successfully amplified the SSRs of R. variegatus, a clam species taxonomically very close to R. philippinarum, from Hong Kong and Jeju Island. We believe that the 11 novel polymorphic SSR developed in this study can be utilized successfully in Manila clam genetic diversity analysis, as well as in genetic discrimination of different clam populations.     

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.     

<Emphasis Type="Italic">In situ</Emphasis> Enclosure Experiment Using a Benthic Chamber System to Assess the Effect of High Concentration of CO<Subscript>2</Subscript> on Deep-Sea Benthic Communities

In order to evaluate the environmental impact associated with sequestration of carbon dioxide in the deep sea, a free fall type field experimental device, the benthic chamber, was developed. In situ experiments to expose deep-sea communities to elevated concentrations of carbon dioxide (average of 20,000 ppm, 5,000 ppm and control) were carried out using this device 3 times, viz., in the winter of 2002 and in the spring and the summer of 2003, in the Kumano Trough at a depth of 2,000 m. In the long-term experiments (about two weeks in winter of 2002 and summer of 2003), the abundance of meiobenthos declined whereas that of bacteria increased under the condition of 20,000 ppm carbon dioxide compared with the control. Among meiofauna, the abundance of foraminifers at the same concentration of carbon dioxide became less than the control even in the short-term (3 days in spring of 2003) experiment, suggesting that organisms with a calcium carbonate exoskeleton are more sensitive to the raised concentration of carbon dioxide. The respiration rate of the benthic community exposed to 20,000 ppm was lower in the early stage of the experiment than in the latter half, whereas it was opposite under the condition of 5,000 ppm. The increase of biological activity in the 20,000 ppm exposure group is probably due to an increase of bacteria adapted to high carbon dioxide concentrations. The present results suggest that the influence of carbon dioxide on the deep-sea benthic ecosystem does not follow a simple, linear relationship with concentration.