Effects of temperature on growth,photophysiology, Rubisco gene expression in <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> and <Emphasis Type="Italic">Karenia mikimotoi</Emphasis>

To explore the effects of temperature changes on dinoflagellate bloom succession in the coastal waters of the East China Sea, changes in the growth, photophysiology, and Rubisco gene expression of Prorocentrum donghaiense and Karenia mikimotoi, two harmful algal species, were investigated at different temperatures (16 to 28°C). The maximal specific growth rate and the maximal mRNA expression of Rubisco gene in P. donghaiense and K. mikimotoi occurred at 20 and 24°C, respectively. The photosynthetic activity of P. donghaiense was generally stable, but K. mikimotoi photosynthesis increased when temperatures rose from 16 to 28°C. The effective photochemical efficiency (F _q ^′ /F _m ^′ ) and the maximal relative electron transfer rate (rETR_max) of K. mikimotoi increased significantly with increasing temperature, and the lowest and highest values occurred at 16 and 28°C, respectively. It seems that P. donghaiense has higher photosynthetic capacity than K. mikimotoi due to its higher F _q ^′ /F _m ^′ , rETR_max, and photosynthetic efficiency (α). However, K. mikimotoi has a higher growth rate than P. donghaiense. These results suggest that the photosynthetic activity and genetic responses of dinoflagellates are species-dependent. It is likely that temperature changes affect species composition during blooms, leading to the observed patterns of bloom succession.     

Temporal changes in the geographic distribution of two clam species <Emphasis Type="Italic">Meretrix lusoria</Emphasis> and <Emphasis Type="Italic">M. petechialis</Emphasis> along the coast of Japan and South Korea

The objectives of this study were to distinguish fossil specimens of Meretrix (Bivalvia, Veneridae) using discriminant score and multivariate analysis, and to examine temporal changes in the geographical distributions of M. lusoria and M. petechialis in Japan and South Korea. Fossil shells of Meretrix were collected from Miyagi, Fukui, Aichi and Kumamoto in Japan, and Gimhae, Seosan, Siheung and Ganghwa-do in South Korea. The outside of the right valve was photographed with a digital camera, and ten characteristics of the shell morphology, such as shell height and shell length, were measured using image analysis software and a digital slide caliper. The discriminant score and multivariate analysis of the shell morphology identified all of the examined fossils as M. lusoria, while living individuals collected from the western coast of South Korea were identified as M. petechialis. These results suggest that the fossil shells excavated from the western coast of South Korea are a different species from the living individuals collected from the same areas. Radioactive carbon isotopic results revealed that the ages of the four fossil shells collected from tidal flats in Ganghwa-do ranged between 3,270 ± 30 and 1,830 ± 30 Cal BP. These results suggest that M. lusoria inhabited the western coast of South Korea until at least 2,000 years ago, but was replaced by M. petechialis during the last 2,000 years. Therefore, we propose that either M. petechialis spats were introduced artificially, or planktonic larvae migrated naturally from China to the western coast of South Korea during the last 2,000 years.     

<Emphasis Type="Italic">Dracograllus trukensis</Emphasis> sp. nov. (Draconematidae: Nematoda) from a seagrass bed (<Emphasis Type="Italic">Zostera</Emphasis> spp.) in Chuuk Islands,Micronesia, Central Western Pacific Ocean

A new species of free-living marine draconematid nematode, Dracograllus trukensis sp. nov., is described based on the specimens collected from the sediments of a intertidal seagrass bed from Chuuk Islands, Micronesia. Dracograllus trukensis sp. nov. differs from other species of the genus by the combination of the following characteristics: the presence of numerous minute spiny ornamented body cuticular annules in both sexes, eight cephalic adhesion tubes inserted on the head capsule in both sexes, the presence of stiff posteriorly directed setae anterior to posterior adhesion tubes in both sexes, the shape (large, elongated, open loop-shaped in male and large, elongated, closed loop-shaped in female) and position (longer ventral arm extending to the first body annule in male) of amphideal fovea, shorter spicule length (34–42 μm), the presence of sexual dimorphism in shape and length of the non-annulated tail terminus, and number of posterior sublateral adhesion tubes (10 in male and 13–15 in female) and posterior subventral adhesion tubes (8–10 in male and 9–11 in female). A comparative table on the biogeographical and ecological characteristics of the species of Dracograllus is presented. This is the first taxonomic report on the genus Dracograllus from Chuuk Islands, Micronesia, central western Pacific Ocean.     

High mortality and poor growth of green mussels, <Emphasis Type="Italic">Perna viridis</Emphasis>, in high chlorophyll-<Emphasis Type="Italic">a</Emphasis> environment

The current study was carried out from May 2014 to April 2015 to estimate the stock status of P. viridis in Marudu Bay. The gonad development was monitored by histological examination, while the population parameters including asymptotic length (L_∞), growth coefficient (K), mortality rate (Z, F and M), exploitation level (E) and recruitment of P. viridis were estimated using the lengthfrequency data. Results of the current study demonstrated that P. viridis in Marudu Bay spawned throughout the year with two major peaks, one in April to May and another one in October to December. The recruitment pattern was continuous with the peak in May to June 2014, which corresponded to the first spawning peak in April. However, no significant recruitment was observed from the second spawning peak due to the difference in spawning timing between male and female populations. The estimated asymptotic length (L_∞), growth coefficient (K), total mortality (Z), natural mortality (M), fishing mortality (F) and growth performance (ф) of P. viridis in Marudu Bay were estimate to be 117 mm, 0.97 yr^-1, 4.39 yr^-1, 1.23 yr^-1, 3.16 yr^-1 and 4.123, respectively. The exponent b of the lengthweight relationship was 2.4 and exploitation level (E) was 0.72. The high mortality, low condition indices and negative allometric of P. viridis in Marudu Bay is caused by a lack of suitable food in the surrounding water.     

Morphological identification of <Emphasis Type="Italic">Alexandrium</Emphasis> species (Dinophyceae) from Jinhae-Masan Bay,Korea

Outbreaks of paralytic shellfish poisoning (PSP) and dense blooms caused by Alexandrium species in Jinhae-Masan Bay, Korea have been nearly annual events for many years. However, excluding some Alexandrium species responsible for PSP, there are no critical reports on the morphology of Alexandrium species in this bay. To identify the Alexandrium species based on detailed morphological features, vegetative cells collected water samples and established by the incubation of resting cysts isolated from sediment trap samples were analyzed. Four species of Alexandrium were identified: Alexandrium affine, A. fundyense, A. catenella, and A. insuetum. Morphological features of these species were basically consistent with those outlined in previous studies. However, the ventral pore and the connecting pore on the sulcal plate, which have been accepted as diagnostic characteristics for the identification of A. fundyense and A. catenella, need to be reevaluated, indicating that useful morphological features for identifying these two species should be recommended to avoid confusion in the classification of species in genus Alexandrium.     

Phylogenetic relationship between symbionts of tubeworm <Emphasis Type="Italic">Lamellibrachia satsuma</Emphasis> and the sediment microbial community in Kagoshima Bay

Vestimentiferan tubeworms acquire their symbionts through horizontal transmission from the surrounding environment. In the present study, we constructed a 16S rRNA gene clone library to investigate the phylogenetic relationship between diverse microbes in the sediment and symbiotic bacteria in the trophosome of the tubeworm, Lamellibrachia satsuma, from Kagoshima Bay, Japan. Two symbiotic bacterial phylotypes belonging to the classes γ- and ε-Proteobacteria were found from this tubeworm trophosome. They were very closely related to the symbionts of several other marine invertebrates. The most predominant bacteria in the sediment were ε-Proteobacteria. A broad diversity of bacteria belonged to non-proteobacterial phyla such as Planctomycetes, Acidobacteria, and Chloroflexi was observed. The presence of sulfur oxidizers (i.e., ε-Proteobacteria and γ-Proteobacteria) and sulfur reducers (i.e., δ-Proteobacteria) may play a significant role in the sulfur cycle in these habitats and provide multiple sources of nutrition to the cold-seep communities. Closely related clones of ε-Proteobacteria symbiont in the species level and of γ-Proteobacteria symbiont in the genus level were found in the surrounding sediment. The similarity of symbiont clones of L. satsuma with other symbionts and free-living bacteria suggests the possibility of opportunistic symbiosis in ε-Proteobacteria and the co-evolution of γ-Proteobacteria having occurred after symbiosis with the tubeworms.     

Measuring chlorophyll <Emphasis Type="Italic">a</Emphasis> concentrations in the Sea of Japan using probe and flow fluorimeters

The spatial variability of chlorophyll a concentrations was studied from the data of two near-shore expeditions and the cruise of the R/V Akademik M.A. Lavrent’ev in October–November 2010 over the northwestern part of the Sea of Japan. The sections across eddies showed a maximum of chlorophyll a at a depth of 40 m. According to the data from the cruise, the chlorophyll a concentration was maximum in the north of the sea and decreased to the south. In parallel, the procedures for chlorophyll a determination were compared for spectrophotometry with a fluorescence probe and a fluorescence flow system. The probe data of chlorophyll a fluorescence showed a high correlation with the chlorophyll a concentrations by spectrophotometry. On the contrary, data on chlorophyll a concentrations from spectrophotometry did not agree with those from the flow system. It was shown that a fluorimeter in the flow system recorded dissolved organic matter along with the chlorophyll a fluorescence.     

Physiological Responses of the Mediterranean Subtidal Alga <Emphasis Type="Italic">Peyssonnelia squamaria</Emphasis> to Elevated CO<Subscript>2</Subscript>

The ecological consequences of ocean acidification are unclear due to varying physiological properties of macroalgae and species-specific responses. Therefore, in the present study, we used a laboratory culture experiment to analyse the eco-physiological responses of the Mediterranean subtidal red alga Peyssonnelia squamaria to CO₂-induced lower pH. Our results showed an increase in the photosynthetic performance and growth rate of P. squamaria, despite the reduction in CaCO₃ content in the low pH treatment. According to our results, we believe that samples exposed to elevated CO₂ could be regulated own nitrogen metabolism to support increased growth rate and it may be down-regulated nitrate uptake. As a result, we hypothesize that P. squamaria may benefit from ocean acidification.     

Species composition and seasonal dynamics of the population density and biomass of the genus <Emphasis Type="Italic">Pyramimonas</Emphasis> (Prasinophyceae) from the Russian waters of East/Japan Sea

The species composition and seasonal dynamics of the population density and biomass of the prasinophycean algae of the genus Pyramimonas were investigated in the Russian waters of the East/Japan Sea. According to literature data and the results of our observations, eight species of the prasinophycean algae were identified, and some of them were new for the Russian waters of the East/Japan Sea as follows: P. aff. amylifera Ñonrad, P. aff. cordata McFadden, Hill et Wetherbee, and P. nansenii Braarud. An analysis of their seasonal dynamics showed that the most conspicuous winter peak of the population density of Pyramimonas species in the Amurskii Bay was clearly distinguishable in February. In winter and early spring, the prasinophycean algae made a considerable contribution of 28 to 60% into the total population density on the background of a relatively low biomass, 1.1–14.4% of the total phytoplankton biomass in the Amurskii Bay. In the Golden Horn Bay, the summer peak of the population density of Pyramimonas species was most intensive in June. In summer, during the period of mass development of the algae of the genus Pyramimonas in the Golden Horn Bay, the prasinophycean algae contributed up to 71% of the total population density and up to 84% of the total microalgal biomass. An increase was noted in the density and biomass of the Pyramimonas species in the polluted waters near the sewage water outlets in the Amurskii and Golden Horn bays.     

Changes in the egg production rate of two copepod species of the genus <Emphasis Type="Italic">Pseudocalanus</Emphasis> in relation to temperature in the White Sea

This study examines changes in egg production rates of two co-occurring Pseudocalanus species, P. acuspes and P. minutus, in response to changing temperatures (0, 3, 6, 9 and 12°C) in the White Sea. The boreal P. acuspes (sample size >650 individuals) increased its reproductive rate gradually across the entire range of temperatures from 0 to 12°C. Significant differences in egg production rates and reaction to temperature were also observed between P. acuspes females incubated during the early vs. the mid-summer season. The sample size of the Arctic P. minutus was significantly smaller due to the low numbers at which this species occurred. However, the results suggest that this species increases its egg production rates from 0 to 3°C from 3.3 to 8.5% dry weight female^–1 day^–1, but at 9° its reproductive rate drops significantly and the animals demonstrate markedly reduced fitness. Our results indicate that closely related co-occurring species, and even different generations within one species, can demonstrate significantly different responses to changes in the physical environment.     

Hybrid <Emphasis Type="Italic">N</Emphasis>-order Lagrangian interpolation Eulerian-Lagrangian method for salinity calculation

The Eulerian-Lagrangian method (ELM) has been used by many ocean models as the solution of the advection equation, but the numerical error caused by interpolation imposes restriction on its accuracy. In the present study, hybrid N-order Lagrangian interpolation ELM (LiELM) is put forward in which the N-order Lagrangian interpolation is used at first, then the lower order Lagrangian interpolation is applied in the points where the interpolation results are abnormally higher or lower. The calculation results of a step-shaped salinity advection model are analyzed, which show that higher order (N=3-8) LiELM can reduce the mean numerical error of salinity calculation, but the numerical oscillation error is still significant. Even number order LiELM makes larger numerical oscillation error than its adjacent odd number order LiELM. Hybrid N-order LiELM can remove numerical oscillation, and it significantly reduces the mean numerical error when N is even and the current is in fixed direction, while it makes less effect on mean numerical error when N is odd or the current direction changes periodically. Hybrid odd number order LiELM makes less mean numerical error than its adjacent even number order LiELM when the current is in the fixed direction, while the mean numerical error decreases as N increases when the current direction changes periodically, so odd number of N may be better for application. Among various types of Hybrid N-order LiELM, the scheme reducing N-order directly to 1st-order may be the optimal for synthetic selection of accuracy and computational efficiency.     

Physiological and biochemical responses of <Emphasis Type="Italic">Prorocentrum minimum</Emphasis> to high light stress

Prorocentrum minimum is a common bloomforming photosynthetic dinoflagellate found along the southern coast of Korea. To investigate the adaptive responses of P. minimum to high light stress, we measured growth rate, and generation of reactive oxidative species (ROS), superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) in cultures exposed to normal (NL) and high light levels (HL). The results showed that HL (800 μmol m^?2 s^?1) inhibited growth of P. minimum, with maximal inhibition after 7–9 days. HL also increased the amount of ROS and MDA, suggesting that HL stress leads to oxidative damage and lipid peroxidation in this species. Under HL, we first detected superoxide on day 4 and H₂O₂ on day 5. We also detected SOD activity on day 5 and CAT activity on day 6. The level of lipid peroxidation, an indicator of cell death, was high on day 8. Addition of diphenyleneiodonium (DPI), an NAD(P)H inhibitor, decreased the levels of superoxide generation and lipid peroxidation. Our results indicate that the production of ROS which results from HL stress in P. minimum also induces antioxidative enzymes that counteract oxidative damage and allow P. minimum to survive.     

New records of three <Emphasis Type="Italic">Paraprionospio</Emphasis> species (Polychaeta: Spionidae) from Korean waters

The spionid polychaete Paraprionospio pinnata (Ehlers 1901) has been widely reported from Korean waters. We examined some specimens belonging to the genus Paraprionospio that had been collected from Korean waters, and identified them as Paraprionospio coora Wilson, 1990, Paraprionospio cordifolia Yokoyama, 2007 and Paraprionospio patiens Yokoyama, 2007, which are new to the fauna of Korea. The present study suggests misidentification of the specimens that have been previously reported as P. pinnata from Korean waters.     

The ecological importance of the dwarf seagrass <Emphasis Type="Italic">Zostera japonica</Emphasis> in intertidal areas on the southern coast of Korea

The intertidal seagrass Zostera japonica, which is distributed in the intertidal and shallow subtidal zones, is exposed to nutrients from over-enriched land-based discharge and storm water runoff before they can be washed out to sea. Despite its ecological importance, only a few studies have examined the ecology and physiology of Z. japonica in northeast Asia. In this study, we investigated the productivity and tissue nutrient contents of above- and below-ground tissues to evaluate the potential role of Z. japonica in carbon capture and as a nutrient sink. The average total, above-, and below-ground productivity per shoot was 0.56, 0.34, and 0.21 mg DW shoot^-1 day^-1, respectively. Annual leaf production was 1.5 times higher than annual below-ground production. Estimated annual whole-plant carbon, nitrogen, and phosphorus incorporation based on shoot production and tissue nutrient contents was 312.8 g C m^-2 y^-1, 25.7 g N m^-2 y^-1, and 1.6 g P m^-2 y^-1, respectively. These values were equivalent to 7.8 × 10³ kg C y^-1, 6.4 × 10² kg N y^-1, and 40 kg P y^-1 for all Z. japonica beds in Geoje Bay. This suggests that Z. japonica meadows can incorporate a considerable amount of carbon, nitrogen, and phosphorus in the intertidal zone. High N:P ratios of above- and below-ground tissues suggest that Z. japonica growth is probably limited by phosphorus availability in the study area.     

Carbon and nitrogen dynamics of the intertidal seagrass, <Emphasis Type="Italic">Zostera japonica</Emphasis>, on the southern coast of the Korean peninsula

Seagrasses require a large amount of nutrient assimilation to support high levels of production, and thus nutrient limitation for growth often occurs in seagrass habitats. Seagrasses can take up nutrients from both the water column and sediments. However, since seagrasses inhabiting in the intertidal zones are exposed to the air during low tide, the intertidal species may exhibit significantly different carbon (C) and nitrogen (N) dynamics compared to the subtidal species. To examine C and N dynamics of the intertidal seagrass, Zostera japonica, C and N content and stable isotope ratios of above- and below-ground tissues were measured monthly at the three intertidal zones in Koje Bay on the southern coast of Korea. The C and N content and stable isotope (δ¹³C and δ¹⁵N) ratios of seagrass tissues exhibited significant seasonal variations. Both leaf and rhizome C content were not significantly correlated with productivity. Leaf δ¹³C values usually exhibited negative correlations with leaf productivity. These results of tissue C content and δ¹³C values suggest that photosynthesis of Z. japonica in the study site was not limited by inorganic C supply, and sufficient inorganic C was provided from the atmosphere. The tissue N content usually exhibited negative correlations with leaf productivity except at the upper intertidal zone, suggesting that Z. japonica growth was probably limited by N availability during high growing season. In the upper intertidal zone, no correlations between leaf productivity and tissue elemental content and stable isotope ratios were observed due to the severely suppressed growth caused by strong desiccation stress.     

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.     

Age determination and growth rate of <Emphasis Type="Italic">Mactra chinensis</Emphasis> (Bivalvia: Mactridae) by external rings and chondrophore growth bands

Age, growth and mortality of Mactra chinensis were investigated during the period from October 2012 to September 2013 in Busan, South Korea. The monthly variation of the marginal index (MI) of the shell and chondrophore showed that the ring of this species was formed once a year during July. We estimated the age of M. chinensis by reading the external rings on the shell and the growth bands of the chondrophore to compare growth parameters between the two growth characters. The age of this species ranged from 0 to 8 years (shell-based age reading) and from 0 to 10 years (chondrophore-based age reading). Based on external rings and growth bands of chondrophore for the same period, the von Bertalanffy growth functions were expressed by the equation, L _t = 101.53[1-exp {-0.15(t + 0.75)}] and L _t = 90.03[1-exp {-0.20(t + 0.50)}], respectively. The likelihood test showed that there was a significant difference in L _∞ (P < 0.001), K (P < 0.001), to (P < 0.001) estimated from non-linear regression between the two growth characters.