Distribution Pattern of Far Eastern Sea Cucumber Apostichopus japonicus in Busse Lagoon (Southern Sakhalin)

We analyzed the distribution pattern and size structure in aggregations of the far eastern sea cucumber (trepang), Apostichopus japonicus in Busse Lagoon (Southern Sakhalin). Trepang populations, which differ in their size composition and population density, have been grouped by their spatial differentiation. It has been shown that in the absence of solid substrates in the lagoon, the animals gather on substrates such as the alga Ahnfeltia tobuchiensis and on clusters of the oyster Crassostrea gigas. It is found that on the southeastern part of the lagoon, trepang makes up an extensive aggregation confined to the bed of Ahnfeltia; the distribution of the holothurians mostly has a random pattern. In the northwestern part of the lagoon, trepang makes up aggregations on oyster and algal beds; the distribution of these aggregations shows a random pattern.     

Vertical Distribution of the Far East Trepang Apostichopus japonicusin Vostok Bay,Sea of Japan

The abundance, weight, and age structure of aggregations of the Far East trepang Apostichopus japonicuswere studied in Vostok Bay, Sea of Japan, during the first part of September 2000. The highest density of aggregations (8.3 ± 0.5 ind/m²) and biomass (131.88 ± 10.50 g/m²) of animals was characteristic for coastal sites at the depth 0.5–1.5 m. With anincrease in depth the biomass and density of A. japonicusreduced 1.5 and 3 (depth 5–6 m), 220 and 830 times (depth 8–15 m) respectively. The ratio of large animals concurrently increased in the aggregation structure. The specifics of the distribution and spatial variability of the aggregation structure are discussed in relation to uncontrolled fishing of holothurians of large sizes.     

New species of pinaceous fossil wood from the Pliocene of southern Primorye (Russian Far East)

New species of the Pinaceae, Abies chavchavadzeae and Piceoxylon ussuriense, are described on the basis of fossil woods from the Pliocene of the Pavlovka lignite field (southern Primorye). For the first time, fossil wood of Abies is reported from the Russian Far East.     

Induction of larval metamorphosis in the sea cucumber Stichopus japonicus by periphitic diatoms

The mass production of juvenile seeds of the sea cucumber,Stichopus japonicus has recently developed by the SeaFarming Center of Saga Prefecture. Methods for the culture ofperiphicic diatoms have been improved. There are three importantsteps in propagating the diatoms. The first step is theenrichment, with the addition of the nutrient salts, undercontrolled light intensity. The second step is washing withhigh pressure seawater and reversal of the plates. The laststep is elimination of copepods, which feed on diatoms, usinga pesticide. Small periphitic diatoms such as Navicula,Amphora, Achnanthes, and Nitzschia are easily culturedat a density of more than one million cells cm^–2, andthese diatoms are able to induce larval metamorphosis andserve as a food source for juvenile sea cucumbers.     

Development and characterization of 45 novel microsatellite markers for sea cucumber (Apostichopus japonicus)

Simple sequence repeat‐enriched library screening and expressed sequence tag database mining were adopted to develop microsatellite markers for sea cucumber (Apostichopus japonicus). Eighty‐three microsatellite loci were selected for polymorphism assessment using 48 individuals. The results showed that 45 novel loci were polymorphic. The number of alleles ranged from two to 16, and the values of observed and expected heterozygosities varied from 0 to 0.9375 and from 0.1135 to 0.9674, respectively. No significant linkage disequilibrium between pairs of loci was found and 26 loci conformed to the Hardy–Weinberg equilibrium. These markers are therefore a potential tool for studies in the population structure and linkage map construction for A. japonicus.     

Reproductive cycle and releasing time for increase of resource of adult sea cucumber Apostichopus japonicus released to seed breeding grounds

The reproductive cycle and releasing time for effective increase of resource of adult sea cucumber (Apostichopus japonicus) have been studied in the west coast of Korea. Adult sea cucumbers collected in Seocheon-gun April 2013 for first release and in the uncontaminated Taean-gun area in the west coast of Korea. Divers monthly collected the specimens in the released area from December 2013 to November 2014 in order to investigate the reproductive cycle of A. japonicas in Taean-gun. Random specimens were dissected to examine the gonadal developmental stages and discharge rates of the guts and gonads. The reproductive cycle of A. japonicus in both sexes are classified into the following six successive stages in the Taean-gun: (1) Stage I (recovery stage from December to February), (2) Stage II (early growing stage from February to March), (3) Stage III (late growing stage from March to April), (4) Stage IV (mature stage from April to July), (5) Stage V (partly spawned stage in July), and (6) Stage VI (spent stage from August to November). The estivation period of this species is from July to October in the Taean-gun region while surface water temperature is approximately 20–25.4°C. Thus, the optimum period to easily harvest them is from October to November in the uncontaminated Taean-gun area. It is also the best releasing time because the recovery stage starts from December to February in the Taean-gun region.     

A comparison of the effects of light intensity on movement and growth of albino and normal sea cucumbers (Apostichopus japonicus Selenka)

Albinism in the sea cucumber Apostichopus japonicus (Selenka) may be a significant biomarker that can be used to obtain information about behaviors and growth of this species. The goal of this study was to compare the behaviors and growth rate between albino and normal A. japonicus. Total light intensity and specific growth rate were used as statistical indices. Diverse light intensities were tested in the laboratory and in the field to identify ecological characteristics of albino A. japonicus. Experimental results revealed that: (1) both albino and normal specimens had negative phototaxis; (2) normal A. japonicus were more responsive to intense light than albino species, and the latter could withstand longer periods of exposure to intense light; (3) normal A. japonicus grew faster than albino specimens; and (4) both normal and albino A. japonicus grew faster in natural light than in the other light intensities tested.     

Virulence genotypes and drug resistance of Helicobacter pylori from Vladivostok, Russia: another feature in the Far East

Helicobacter pylori in Vladivostok, Far Eastern Russia, was investigated during 2004 to 2009. The genotype cagA(+) vacA(+) (s1/m1 or m2) accounted for 74.7%, with cagA(-) vacA(+) (s2/m2) at 11.2%. The CagA EPIYA type was mainly Western ABC, with minor types (ABCCC and novel AAABC) or non-Western/non-East Asia type (AB). Regarding drug resistance, metronidazole resistance was the highest, with a marked decrease in 6 years (from 71.4% to 30.8%); in contrast, levofloxacin and clarithromycin resistance increased. The data indicate that in Vladivostok, H. pylori was mainly the Western (not East Asian) type and dynamic changes in drug resistance occurred during 6 years.     

Nervous system development of the sea cucumber Stichopus japonicus

The nervous system development of the sea cucumber Stichopus japonicus was investigated to explore the development of the bilateral larval nervous system into the pentaradial adult form typical of echinoderms. The first nerve cells were detected in the apical region of epidermis in the late gastrula. In the auricularia larvae, nerve tracts were seen along the ciliary band. There was a pair of bilateral apical ganglia consisted of serotonergic nerve cells lined along the ciliary bands. During the transition to the doliolaria larvae, the nerve tracts rearranged together with the ciliary bands, but they were not segmented and remained continuous. The doliolaria larvae possessed nerves along the ciliary rings but strongly retained the features of auricularia larvae nerve pattern. The adult nervous system began to develop inside the doliolaria larvae before the larval nervous system disappears. None of the larval nervous system was observed to be incorporated into the adult nervous system with immunohistochemistry. Since S. japonicus are known to possess an ancestral mode of development for echinoderms, these results suggest that the larval nervous system and the adult nervous system were probably formed independently in the last common ancestor of echinoderms.     

Amino acid sequence of the coelomic C globin from the sea cucumber Caudina (Molpadia) arenicola.

The sequence of a globin from a marine invertebrate, the sea cucumberCaudina (Molpadia) arenicola (Echinodermata), is reported. This globin, chain C, is one of four major globins found in coelomic red cells in this organism and is the second to be sequenced. Chain C consists of 157 residues, is amino-terminally acetylated, and has an extended amino-terminal region. This globin shares a 60% sequence identity with the other sequencedC. arenicola globin, D chain (Mauriet al., Biochem. Biophys. Acta1078, 63–67, 1991), but has a 93.6% identity with a globin from another sea cucumber,Paracaudina chilensis (Suzuki,Biochem. Biophys. Acta,998, 292–296, 1989).     

A framework for providing scientific advice for the management of new and developing invertebrate fisheries

A framework is developed for the provision of scientific advice to support the management of new and developing marine invertebrate fisheries. These fisheries often occur on species for which little biological or exploitation information is available. The framework explicitly endorses the precautionary approach to fisheries management and research. Three general management strategies (size/sex limits, regulation by total allowable catch, control of the exploitation rate) and their needs for supporting scientific information are identified. The significance of spatial pattern, and of recognizing the need for different approaches to obtain scientific information and to manage sedentary benthic and mobile pelagic species, is a central theme. Three 'phases' are proposed to obtain the necessary scientific information: (a) Phase 0, 'collecting existing information', consisting of syntheses of available biological and fisheries information on the target (and similar) species, leading to formulation of potential management strategies; (b) Phase 1, 'collecting new information', to obtain the essential information that is lacking or insufficient from the Phase 0 analysis, and to evaluate alternative management strategies and propose regulatory actions; and (c) Phase 2, 'fishing for commerce', to implement the chosen management actions and to monitor fishing operations, so as to increase the information base available to refine the results from previous phases. Phase 1 activities may consist of surveys, site-specific depletion experiments and studies to obtain biological information, and development of experimental management areas to test different exploitation rates. A strategy that includes establishing reserve areas recognises the inherent uncertainties associated with developing fisheries and provides a buffer against mistakes or 'surprises'; it also provides control areas to compare stock productivity in fished and unfished locations. The application of this framework to a developing sea cucumber fishery in British Columbia is presented as an example. Throughout, strong interaction and collaboration among science, management, and stakeholders is crucial to the provision of scientific advice for precautionary management of new invertebrate fisheries.     

High abundance of Myriochele fragilis Nilsen & Holthe, 1985 (Polychaeta: Oweniidae) in the deep sea of the Eastern Mediterranean

An exceptionally high abundance of Myriochele fragilis Nilsen & Holthe, 1985 (Polychaeta: Oweniidae) is reported for the Ierapetra Basin in the Eastern Mediterranean Sea from depths of 4159 and 4260 m, representing the first record outside the Norwegian and Arctic Sea and extending the depth range by more than 1600 m. Specimens are described and compared to closely related species.     

Purification and Characterization of a Cathepsin L-Like Enzyme from the Body Wall of the Sea Cucumber Stichopus japonicus

Cathepsin L-like enzyme was purified from the body wall of the sea cucumber Stichopus japonicus by an integral method involving ammonium sulfate precipitation and a series of column chromatographies on DEAE Sepharose CL-6B, Sephadex G-75, and TSK-GEL. The molecular mass of the purified enzyme was estimated to be 63 kDa by SDS–PAGE. The enzyme cleaved N-carbobenzoxy-phenylalanine-arginine

7-amido-4-methylcoumarin with K _m (69.92 μM) and k _cat (12.80/S) hardly hydrolyzed N-carbobenzoxy-arginine-arginine 7-amido-4-methylcoumarin and L-arginine 7-amido-4-methylcoumarin. The optimum pH and temperature for the purified enzyme were found to be 5.0 and 50 °C. It showed thermal stability below 40 °C. The activity was inhibited by sulfhydryl reagents and activated by reducing agents. These results suggest that the purified enzyme was a cathepsin L-like enzyme and that it existed in the form of its enzyme-inhibitor complex or precursor.     

Stichopin-containing nerves and secretory cells specific to connective tissues of the sea cucumber

Stichopin, a 17-amino acid peptide isolated from a sea cucumber, affects the stiffness change of the body-wall catch connective tissues and the contraction of the body-wall muscles. The localization of stichopin in sea cucumbers was studied by indirect immunohistochemistry using antiserum against stichopin. Double staining was performed with both stichopin antiserum and 1E11, the monoclonal antibody specific to echinoderm nerves. A stichopin-like immunoreactivity (stichopin-LI) was exclusively found in the connective tissues of various organs. Many fibres and cells with processes were stained by both the anti-stichopin antibody and 1E11. They were found in the body-wall dermis and the connective tissue layer of the cloacae and were suggested to be connective tissue-specific nerves. Oval cells with stichopin-LI (OCS) without processes were found in the body-wall dermis, the connective tissue sheath of the longitudinal body-wall muscles, the connective tissue layer of the tube feet and tentacles, and the connective tissue in the radial nerves separating the ectoneural part from the hyponeural part. Electron microscopic observations of the OCSs in the radial nerves showed that they were secretory cells. The OCSs were located either near the well-defined neural structures or near the water-filled cavities, such as the epineural sinus and the canals of the tube feet. The location near the water-filled cavities might suggest that stichopin was secreted into these cavities to function as a hormone.     

Expression,purification, and characterization of arginine kinase from the sea cucumber Stichopus japonicus

The arginine kinase gene of sea cucumber Stichopus japonicus was cloned and inserted into the prokaryotic expression plasmid pET-21b. The protein was expressed in a soluble and functional form in Escherichia coli and purified by Blue Sepharose CL-6B, DEAE-32, and Sephadex G-100 chromotography with a final yield of 83 mgL(-1) of LB medium. The specific activity, electrophoretic mobility, and isoelectric focusing were all identical with those of arginine kinase that was purified from sea cucumber muscle. The fluorescence emission spectrum of arginine kinase had a maximum fluorescence at a wavelength of 330 nm upon excitation at 295 nm. These results are the first report of this purified protein.     

Heavy metal accumulation in tissue/organs of a sea cucumber, Holothuria leucospilota

Holothuria leucospilota Brandt, the large black sea cucumber, is a non-selective deposit feeder, and is commonly found in the bottom of shallow waters in Hong Kong, where the sediments are often polluted with heavy metals. This study was designed to test the possibility of heavy metal accumulation by the sea cucumber at two sites in Hong Kong. Atomic absorption spectrophotometer was used to measure Cu and Zn concentrations in various tissue/organs of the animal as well as in the sediments. The result indicated that H. leucospilota accumulated zinc in the longitudinal muscle bands (97.27–98.07 ppm in dry weight) and in the respiratory tree (83.92–89.64 ppm in dry weight). Copper concentrations in these two organs were much lower than that of zinc. After the animals were kept in the aquarium without sediment for40 days, zinc concentration of the longitudinal muscle and respiratory tree decreased by 48% and 39%respectively whereas copper concentration remained unchanged. The concentrations of zinc and copper in the sediment at the two sites differed significantly but the metal level in the animals from the two sites were similar, suggesting that this sea cucumber was not an ideal bioindicator of heavy metal pollution in the sediment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Allele-specific PCR genotyping of the HSP70 gene polymorphism discriminating the green and red color variants sea cucumber （Apostichopus japonicus）

Color variation is a well-known feature of sea cucumbers (Apostichopus japonicus),which are classified into three groups based on their colors of red,green and black.It is also one of the most important traits related to how they taste,and it thereby affects their market price.Attempts were made to identify single-nucleotide polymorphisms (SNPs) and to analyze differences associated with SNP genotypes between green and red color variants using HSP70 as the target gene.The HSP70 gene,which is found universally in organisms from bacteria to humans,is one of the most evolutionarily conserved genes and the most widely studied biomarker of stress response.DNA fragments of 1074 bp covering a partial sequence of the sea cucumber HSP70 gene,were amplified from both red and green variants,and subsequently analyzed for the presence of SNPs.Twenty-seven polymorphic sites in total,including heterozygous sites,were observed.Of these,six sites were found to be significantly different SNP genotypes between green and red variants.Furthermore,PCR with an internal primer designed to include an allelespecific SNP at the 3' end (site 443) showed differentiation between the two variants,100％ and 4.2％ amplification in green and red variants,respectively.The validated SNPs may serve as informative genetic markers that can be used to distinguish variants at the early developmental stage,prior to color differentiation.     

Isolation and characterization of a sea cucumber fucoidan‐utilizing marine bacterium

Aims: To isolate a fucoidan‐utilizing strain from seawater for sea cucumber fucoidan degradation. Methods and Results: The utilization of sea cucumber fucoidan was monitored by H₂SO₄–phenol assay for neutral sugar. The bacterium CZ1127 was isolated from seawater and shown to have a relatively large maximum fucoidan‐utilizing rate of 81·5%. CZ1127 was confirmed to belong to the family Flavobacteriaceae by 16S rDNA and physiological analyses. This strain has an ability to utilize fucoidans extracted from various sea cucumbers to different degrees. Both extracellular and intracellular enzymes of CZ1127 could degrade sea cucumber fucoidan, as confirmed by high‐performance size exclusion chromatography. The M_r of sea cucumber fucoidan could be reduced from 792·6 kDa to at least 3·7 kDa by the crude intracellular enzyme of this strain. Conclusions: The marine bacterial strain CZ1127, which belongs to the family Flavobacteriaceae, was found to utilize various sea cucumber fucoidans and furthermore showed promise in sea cucumber fucoidan enzymatic degradation and oligosaccharide preparation. Significance and Impact of the Study: The finding of a novel source can be applied in sea cucumber fucoidan enzymatic degradation. Furthermore, it is the first definite report of a bacterial strain that can utilize the fucoidans from various sea cucumbers.     

Isolation and Characterization of Twenty Microsatellite Loci in Japanese Sea Cucumber (Stichopus japonicus)

Twenty microsatellite markers were first developed from the Japanese sea cucumber Stichopus japonicus using an enrichment protocol. Of the 20 microsatellite loci, 19 loci were polymorphic in the population examined. At these polymorphic loci, the number of alleles per locus varied from 2 to 15, and the observed heterozygosities ranged from 0.03 to 0.97, which is considerably higher than those previously found for allozymes. The high variability of the microsatellite markers identified in this study will make them excellent tools for genetic analyses of S. japonicus.