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.     

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.     

Characterization of the coccoid cyanobacterium <Emphasis Type="Italic">Myxosarcina</Emphasis> sp. KIOST-1 isolated from mangrove forest in Chuuk State,Federated States of Micronesia

Mangrove forests are known to be inhabited by diverse symbiotic cyanobacterial communities that are capable of N₂ fixation. To investigate its biodiversity, root sediments were collected from a mangrove forest in Chuuk State, Federated States of Micronesia (FSM), and an entangled yellow-brown coccoid cyanobacterium was isolated. The isolated cyanobacterium was reproduced by multiple fission and eventually produced baeocytes. Phylogenetic analysis revealed that the isolate was most similar to the genera Myxosarcina and Chroococcidiopsis in the order Pleurocapsales. Compositions of protein, lipid and carbohydrate in the cyanobacterial cells were estimated to be 19.4 ± 0.1%, 18.8 ± 0.4% and 31.5 ± 0.1%, respectively. Interestingly, total fatty acids in the isolate were mainly composed of saturated fatty acids and monounsaturated fatty acids, whereas polyunsaturated fatty acids were not detected. Based on the molecular and biochemical characteristics, the isolate was finally classified in the genus Myxosarcina, and designated as Myxosarcina sp. KIOST-1. These results will contribute to better understanding of cyanobacterial biodiversity in the mangrove forest in FSM as well as the genus Myxosarcina, and also will allow further exploitation of its biotechnological potential on the basis of its cellular characteristics.     

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.     

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.     

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.     

Feeding,egg production,and respiration rate of pteropods <Emphasis Type="Italic">Limacina</Emphasis> in Arctic seas

The feeding, egg production, and respiration rate of the dominant pteropod Limacina helicina have been studied in Russia’s Arctic seas. The sinking rates of fecal pellets and dead individuals have been measured to estimate their role in vertical carbon flux. As has been shown, the rate of ecophysiological processes taking place in the pteropods is higher than that of copepods, the main consumers of phytoplankton. The gut pigment content in Limacina (3084 ng ind^–1 as a maximum) was two orders of magnitude higher than in copepods. The egg production rate in Limacina even without feeding reached 4000 eggs ind^–1 versus 350–450 egg ind^–1 typical of the dominant copepods even with excess food. A close correlation between the pteropod feeding rate and individual body weight was observed for Limacina rather than a correlation with food concentration. The experimentally estimated sinking rate of Limacina fecal pellets was 270 m day^–1, higher than for most copepods. The sinking rate of dead pteropods reaches 2000 m day^–1. According to the literature, discarded mucous feeding nets sink at a rate of 80 to 1080 m day^–1. Evidently, pteropods play a significant role in biogeochemical cycles by accelerating sedimentation. High rates of all studied processes suggest that Limacina are an important component of plankton communities and play the most important role in trophodynamics at sites of their accumulation.     

The reproductive cycle of the humbug damselfish <Emphasis Type="Italic">Dascyllus aruanus</Emphasis> according to the changes in lunar phase in Micronesia

The moon is known to be an environmental factor that controls the reproductive cycle of fish, and fish have evolved a variety of reproduction patterns depending on the lunar phase. In this study, we examined the relationship between lunar phases and the reproductive cycle of the humbug damselfish Dascyllus aruanus inhabiting Weno Island, Chuuk Lagoon, Micronesia. We divided the one-month lunar cycle into eight phases, and measured the features of moonlight (peak wavelength and intensity) and indicators of fish maturity [gonadosomatic index (GSI) and sex hormones] in relation to the eight lunar phases. In addition, we investigated the daily rhythms of sex hormones in fish ovaries during the full moon phase. The results showed that the peak wavelength of moonlight was 430 nm (blue wavelength region) regardless of the lunar phase, and that moonlight intensity was highest during a full moon at 02:00. Furthermore, the GSI and sex hormones were both higher around the full moon phase. These findings support the hypothesis that humbug damselfish spawn once a month and that this event occurs at full moon, which is the phase of the moon with the strongest intensity. Based on these findings, we predict that blue wavelength, the dominant wavelength of moonlight, is one of the environmental factors influencing the monthly spawning of D. aruanus.     

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.     

<Emphasis Type="Italic">S</Emphasis>-wave velocity structure and tectonic implications of the northwestern sub-basin and Macclesfield of the South China Sea

Based on the optimum P-wave model, the S-wave velocity structure of a wide angle seismic profile (OBS2006-1), across the northwestern sub-basin (NWSB) and the Macclesfield, is simulated by a 2-D ray-tracing method. The results indicate the S-wave velocities in the upper and lower crust of the NWSB are 3.2–3.6 km/s and 3.6–4.0 km/s, with Vp/Vs ratios of 1.82–1.88 and 1.74–1.82, respectively, which reflect typical oceanic crust characteristics. The S-wave velocity in the upper crust of the NWSB is a little higher in the NNW segment than that in the SSE segment, while the lateral variation of Vp/Vs ratio is in the opposite. We suggest that the NWSB might have experienced asymmetrical magma flows during sea floor spreading, which may have blurred the magnetic anomaly lineation. The comparison of S-wave velocities along the northern margin of the SCS shows that the west section is different from the east section, and the northwestern margin has a non-volcanic crust structure. The S-wave structures and P-wave velocity models along the northern margin, Macclesfield and Reed Bank show that the Macclesfield might have a conjugate relationship with the Reed Bank.     

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.     

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.     

Spatial Distribution and Seasonal Dynamics of the Chlorophyll <Emphasis Type="Italic">a</Emphasis> Concentration in the Sea of Azov Based on MERIS Images

The chlorophyll a concentration (Cchl a) in the Sea of Azov is estimated by the two-band NIR-red algorithm [34] from MERIS images for 2002–2012. The sea-truth spectrophotometric measurements and MERIS remote estimates of Cchl a are compared. The monthly average Cchl a values are mapped from MERIS data for its lifetime for the first time. The features of the spatiotemporal distribution of Cchl a are ascertained. Differences between the seasonal dynamics of Cchl a in the Sea of Azov according to the literature data and the dynamics derived from MERIS data are found, namely: the summer–autumn phytoplankton growth period is longer than the spring period and is characterized by higher Cchl a values throughout the water area.     

Annual gametogenesis and reproductive effort of the limpet <Emphasis Type="Italic">Cellana grata</Emphasis> (Gould, 1859) (Gastropoda: Nacellidae) in a rocky intertidal beach at Ulleungdo Island off the east coast of Korea

Widely distributed from the northern coast of Vietnam to the northern Japan, the limpet Cellana grata (Gould, 1859) occurs commonly on the south and east coasts of Korea. Despite their wide distribution range, few studies have investigated the annual gametogenesis and reproductive effort of C. grata. In an attempt to understand the reproductive physiology of the limpet, we investigated the annual gametogenesis and reproductive effort of C. grata from Ulleungdo Island off the east coast of Korea. Histology revealed that the gonial mitosis commenced in January, as the female exhibited small oogonia (10–40 μm) in the follicle. From March to June, the oocyte size increased dramatically, and fully mature eggs (110–170 μm in diameter) appeared in early summer. First spawning males and females were observed in July, as the surface seawater temperature (SST) reached 22.1°C. The spawning male and females could be observed until the end of December. Gonad somatic index (GSI), a ratio of gonad mass to the total tissue weight, of the male ranged from 0.6 (April) to 17.9 (July), while the female GSI varied from 1.0 (February) to 18.3 (July). GSI of male and female declined rapidly from July to August, suggesting that the major purse of the spawning at the study site was between July and August. Our study suggested that the commercial catch of C. grata during July and August must be suspended at Ulleungdo Island, in order to protect the spawning limpets, which enhances C. grata recruitment and the population.     

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.     

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.     

Reproductive biology of blood cockle <Emphasis Type="Italic">Anadara granosa</Emphasis> (Bivalvia: Arcidae) in the northern region of the Strait of Malacca

A study on the reproductive cycle of the blood cockle Anadara granosa (Bivalvia: Arcidae) was conducted at three different areas in the northern region of the Strait of Malacca. A total of 1,920 samples of adult A. granosa (38–71 mm length) were collected from June 2009 until September 2010. Qualitative techniques (gonadal microscopic fresh smear test and histology analysis) as well as quantitative techniques (analysis of condition index and gonadal index) were used to predict monthly gonadal development stages of A. granosa. The gonadal index of A. granosa from Banda Aceh (Indonesia) (r = 0.469, P > 0.05) and Pulau Pinang (Malaysia) (r = 0.123, P > 0.05) did not show any correlation to their condition index, whereas the gonadal index of A. granosa from Lhokseumawe (Indonesia) (r = 0.609, P < 0.05) showed moderate positive correlation to the condition index. During the 16 month sampling period, four reproductive cycles were observed: each from three to six months. The process of releasing gametes is termed dribble spawning, and is the same in all populations. The principle component analysis (PCA) indicated that A. granosa reproduction was affected by interaction between internal physiological factors and indigenous environmental factors. In all sampling areas, phytoplankton density played a key role in the reproductive cycle in A. granosa. Information on the reproductive biology of this species is essential for species management and to improve the sustainability practices of the fisheries industry. These findings will provide basic information on the biology of the blood cockle A. granosa for stock management in the region.     

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.