Spawning induced by cubifrin in the Japanese common sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

The neuropeptide cubifrin-I and its derivative cubifrin-L have recently been shown to be potent substances that induce in vitro oocyte maturation and spawning in the Japanese common sea cucumber Apostichopus japonicus. We have examined the reproductive behavior of A. japonicus in vivo following the injection of cubifrin-L into the body cavity with the aim of determining the practical value of using this peptide to induce spawning in the hatchery setting. The responsiveness of ovarian fragments with oocytes >155 μm in diameter to cubifrin-L in vitro was well correlated with the spawning success induced in vivo by a cubifrin-L injection. Mature sea cucumbers injected with cubifrin-L displayed sequential reproductive behaviors, which comprised climbing up the side wall of the tank toward the water surface, waving of the head, and shedding of gametes. Gamete shedding started about 60 and 80 min after the injection of cubifrin-L in males and females, respectively, and was completed almost simultaneously in the two sexes about 2 h post-injection. Repeated injections of cubifrin-L at intervals of about 10 days successfully induced multiple spawnings in males and females. These results demonstrate that cubifrin-L can be used as an effective inducer of spawning in Japanese sea cucumber cultivation.     

Seasonal distribution pattern of adult sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> (Stichopodidae) in Yoshimi Bay,western Yamaguchi Prefecture,Japan

To examine the distribution pattern of adult Apostichopus japonicus, approximately monthly surveys were conducted during a 2-year period in the subtidal zone of a harbor area in Yoshimi Bay, western Yamaguchi Prefecture. Results showed a clear seasonal change in distribution pattern: many animals were distributed on the seabed and the lower part of a jetty during winter to spring, whereas most animals occurred on the upper part of a jetty during summer to autumn, namely, the aestivation season. Such change is considered to be due to the migration of animals within the present study site. Y. Yamana is a Research Fellow of the Japan Society for the Promotion of Science.     

Single spaghetti tagging as a high-retention marking method for Japanese common sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Populations of sea cucumbers, including the Japanese common sea cucumber Apostichopus japonicus, have been seriously depleted worldwide due to overfishing. Mark–recapture study is an efficient means of collecting ecological data. However, the use of such a method in sea cucumbers is difficult because they lack hard tissues in the body wall. Here we tested the viability of various tagging methods on A. japonicus. First, we applied conventional tags using four different methods [single spaghetti (T-bar) tagging, double spaghetti tagging, ribbon tagging, and Atkins tagging] to ten individuals per method in aquaria for 14 days. Of the methods used, single spaghetti tagging had the highest retention rate. Then we examined the retention rate of single spaghetti tags on ten individual sea cucumbers for up to approximately 6 months in rearing conditions. The single spaghetti tagging method showed a retention rate of 100% over at least 7 days, and 50% of the tags remained embedded after 56 days. The longest duration of tag retention was 174 days, at which time the experiment was terminated. These results indicate that single spaghetti tagging is reliable for both short- and longer-term studies, making it a useful tool for ecological and conservation studies in sea cucumbers.     

Effect of salinity and temperature on salinity tolerance of the sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Sea cucumber Apostichopus japonicus juveniles acclimated to different environmental conditions (23, 25, and 27°C combined with 25, 30, and 35 psu) were assessed for tolerance to increasing and decreasing levels of salinity at a rate of 2 psu h⁻¹. They were also tested for the LS₅₀ (median lethal salinity) when transferred directly into a series of higher salinity (32–46 psu) and lower salinity (9–25 psu). The CSMax (critical salinity maximum), CSMin (critical salinity minimum), USTL (upper salinity tolerance limit), and LSTL (lower salinity tolerance limit) were positively correlated to the acclimated salinity level but negatively correlated to temperature. The CSMax of A. japonicus was 6.2–10.0 psu higher than the USTL, and the CSMin was 5.5–8.5 psu lower than the LSTL, indicating that gradual changes in salinity resulted in the wide range of salinity tolerance that was observed, but that abrupt changes in salinity resulted in the narrow range of tolerance. Two-way analysis of variance revealed that salinity and temperature had a significant effect on 50% CSMax, 50% CSMin, USTL, and LSTL (P < 0.001). The information obtained in this study will be valuable for the further development of the sea cucumber aquaculture industry in China.     

Laboratory observations of habitat selection in aestivating and active adult sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

In order to verify which factors affect habitat selection for aestivating and in the active adult Apostichopus japonicus, animals were tested for their selection of attachment site in an experimental device (1-m pipes) in which the perceived environmental stimuli (light intensity, degree of contact with a hard surface, geotaxis) varied depending on the attachment site. During the aestivating season, the animals showed a strong selection for attachment sites during the daytime and nighttime; they also showed positive stereotaxis (thigmotaxis), negative phototaxis, and negative geotaxis. The results suggest that (1) habitats are not suitable for the aestivating adult A. japonicus unless these three environmental requirements are satisfied.     

Induced thermotolerance and expression of heat shock protein 70 in sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

ABSTRACT:   Thermal limits, induced thermotolerance and the expression of heat shock protein 70 (Hsp70) in an echinoderm Apostichopus japonicus were studied. The sublethal and lethal temperatures for the juveniles were 30 and 34°C, respectively; a previous sublethal heat shock exposure (30°C, 2 h) could increase the survival rates of the sea cucumbers when they were exposed to 34°C. This induced thermotolerance could last for at least 2 days. Levels of Hsp70 increased substantially after sublethal heat shock exposure and linearly decreased with time. This result indicated that a close relationship existed between the induction of thermotolerance and the levels of Hsp70 in A. japonicus.     

Feeding activity rhythm in Japanese sea catfish <Emphasis Type="Italic">Plotosus japonicus</Emphasis>

To examine the feeding activity characteristics of Japanese sea catfish Plotosus japonicus, access to feed box (feeding activity) and the number of food pellets consumed (food intake) were recorded under laboratory conditions. When fed ad libitum under a 12:12 h light–dark (LD) cycle, all fish exhibited feeding activity and consumed food during the dark period. Feeding activity increased in the presence of food, and this increased activity level continued for several days after food removal. During restricted food availability within the light period of the LD cycle, seven of nine fish exhibited food-anticipatory activity (FAA) both before and after the 12:00 food-restricted time. This FAA persisted under constant light conditions (in nine of nine fish). The average circadian free-running period was 24 h. These results suggest that P. japonicus exhibits a nocturnal feeding activity rhythm, which may be synchronized by a food-restricted time.     

Effects of heat-shock selection during pelagic stages on thermal sensitivity of juvenile sea cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka

The effects of heat-shock during pelagic stages on growth and survival of juvenile sea cucumber Apostichopus japonicus were investigated to test the possibility to acquire high thermal tolerance individuals after the heat-shock selection. Larvae at the stages of blastula, gastrula, and auricularia were heat-shocked at selected temperatures (21.5, control; 26, 28, and 30°C) for 45 min and returned to 21.5°C for continuous rearing. There was a significant difference in thermal sensitivity among different developmental stages, and thermal tolerance of the larvae was correlated with the expression of heat shock protein 70 (Hsp70). Juveniles after the heat-shock selection at pelagic stages showed higher induced thermotolerance than those without heat-shock. Therefore, heat-shock application at early pelagic development stages is potentially a useful way to select high-thermotolerance variety of the sea cucumbers.     

Natural growth of juveniles of the sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>: studying juveniles in the intertidal habitat in Hirao Bay,eastern Yamaguchi Prefecture,Japan

In order to determine the population dynamics of juvenile Apostichopus japonicus, successive surveys were conducted in the stony intertidal zone of Hirao Bay, Seto Inland Sea. The juveniles of the green and black types of this species grew well from January to June. Before this high growing season, zero year old juveniles were too small to be readily detected by visual observations during field surveys. During the high water temperature season from August to November, juveniles over 1 year of age estivated underneath rocks. They awakened from their estivation in November, and it took another month before they could eat and discharge. The juveniles mainly migrated from the intertidal zone before they reached 3 years of age. In the present work, three patterns of juvenile growth were found. It is suggested that these differences in juvenile growth are mainly caused by differences in their start times for growth initiation when they are zero years old.     

Microbial community in a multi-trophic aquaculture system of <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>, <Emphasis Type="Italic">Styela clava</Emphasis> and microalgae

An integrated multi-trophic aquaculture (IMTA) system consisting of the ascidian Styela clava and the sea cucumber Apostichopus japonicus with microalgae was grown in a seawater mesocosm. Microbial populations in the water, sediment and shelters were monitored over time. The composition of the water microbial community in the IMTA system did not differ significantly from that of the traditional culture system without ascidians. Bacterial populations in the water, sediment and shelters were low in the traditional A. japonicus aquaculture system, and lower in the IMTA system. The number of Vibrios in the water, sediment and shelters in the IMTA system was significantly lower than in the traditional culture (p < 0.05), while the quantity of Bacillus was higher in the sediment and shelters. All of these results indicate that the IMTA system effectively inhibited the growth of harmful bacteria, an important positive function in the health of the culture. The bacterial diversity index in the water in the IMTA system was between those of the baited and non-baited traditional aquacultures and was slightly reduced in the shelters. Over time, the bacterial diversity index in the sediment gradually dropped, especially in summer and autumn, when it was significantly lower than in the traditional culture. In winter, the diversity index increased to some extent, approaching that of the traditional culture.     

Effect of yeast polysaccharide on the immune function of juvenile sea cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka under pH stress

In the present study, we evaluated the effect of yeast polysaccharide (YSP) on the immune function of Apostichopus japonicus at different pH levels. Juvenile sea cucumbers were fed artificial feed containing 0.5% YSP. Phagocytic activity and activity of lysozyme (LSZ), acid phosphatase (ACP) and alkaline phosphatase (ALP) were measured under pH stress after 24 h. The results showed that the phagocytic index, the activity of LSZ and ACP in the intestines increased at pH 10.0, and ACP activity in the body wall decreased. The phagocytic rate and ALP activity in the intestines were not different from the control group. The phagocytic index was also not different from the control group at pH 5.5. The activity of LSZ and ACP in the intestines and the activity of ALP increased, while the activity of ACP in the body wall decreased. The phagocytic rate of YSP group increased and the phagocytic rate of control group decreased. Thus, this study indicated that YSP can modulate the non-specific immunity of A. japonicus, and the reasonable dose of YSP might be used as the immunostimulants in sea cucumbers farming under pH stress.     

Effects of dietary rhubarb, <Emphasis Type="Italic">Bacillus cereus</Emphasis>, yeast polysaccharide,and florfenicol supplementation on growth,intestinal morphology,and immune responses of sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis>)

A 60-day feeding trial was conducted to investigate the effects of dietary Chinese herbal medicine (rhubarb, Rheum officinale Bail), probiotic (Bacillus cereus BC-01), prebiotic (yeast polysaccharide), and antibiotic (florfenicol) supplementation on the growth, intestinal morphology, and immune responses of juvenile sea cucumbers (Apostichopus japonicus Selenka; initial weight, 4.67 ± 0.06 g). The control treatment was fed only the basal diet (CT treatment), whereas four other treatments were fed the basal diet supplemented with rhubarb (RH treatment), B. cereus BC-01 (BC treatment), yeast polysaccharide (PO treatment), or florfenicol (FL treatment). Groups of ten sea cucumbers were housed in separate tanks, and each diet was fed to five randomly selected groups. The specific growth rates of the individuals in the RH and BC treatments were significantly higher than those in the CT, PO, and FL treatments. The apparent digestibility coefficient of crude protein in the RH and BC treatments was significantly higher than that of crude protein in the CT and PO treatments. The alkaline phosphatase activity of the coelomocytes in the RH and BC treatments was significantly higher than that in the CT and FL treatments. Coelomocytes in the PO treatment had the highest acid phosphatase activity. Higher than average lysozyme activity was observed in the PO and FL treatments. Fold and enterocyte heights in the mid-intestinal tract of sea cucumbers in the CT and RH treatments were lower than those in the BC, PO, and FL treatments. Microvillus height was greatest in the RH treatment and lowest in the FL treatment. Scanning electron microscopy revealed remarkable atrophy of microvilli and apoptosis of intestinal epithelial cells in the FL treatment. These results suggest that dietary rhubarb, B. cereus, and yeast polysaccharide supplementation can improve the health of sea cucumbers, whereas long-term florfenicol supplementation may have a negative impact on their intestinal healthy and immune response of sea cucumbers.     

Japanese nephropid lobster <Emphasis Type="Italic">Metanephrops japonicus</Emphasis> lacks zoeal stage

ABSTRACT:   Larvae of the Japanese nephropid lobster Metanephrops japonicus hatched in the laboratory were reared at 15°C, and the development and feeding were observed. All larvae hatched at the 'prezoea stage' with no natatory setae on the exopodite of the pereiopods. Without feeding, 50% of prezoea molted into the megalopa stage, having small buds as the exopodite, within 1 h and all molted within 22 h. The megalopa fed with Artemia nauplii, shrimp meat and pelleted food molted into the first juvenile stage with no exopodite after approximately 17 days. The average carapace lengths of prezoea, megalopa and the first juvenile stage were 3.2, 3.6 and 4.4 mm, respectively. The survival rate from hatching to the first juvenile stage was high (90–100%). This lobster may be the only known nephropid species with no zoeal stage.     

Bathymetric distribution of beni-zuwai crab <Emphasis Type="Italic">Chionoecetes japonicus</Emphasis> in the northern part of the Sea of Japan

Surveys of beni-zuwai crab were conducted to clarify the bathymetric distribution and fecundity of beni-zuwai crab populations without the effects of bias due to fisheries. The results were compared with previous ones obtained at the heavily exploited Yamato Bank. Two sites in the northern part of the Sea of Japan, off southwest Hokkaido (200–2500 m deep) and West-Tsugaru Basin (300–1900 m deep), both of which are experiencing low exploitation levels, were surveyed in August to September, 2007. There was an abundance of morphologically immature crabs of both sexes in the depth range 1800–2100 m, and a few occurred at depths >2200 m. Immature crabs of <40 mm carapace width were found to migrate downslope to depths of 1800–2100 m and then to migrate upslope during molting for males and after maturity molting for females. Although data were lacking for depths >2000 m at the West-Tsugaru Basin, these relationships were significant for the pooled data of both localities, and they were similar to the results at the Yamato Bank. Clutch fullness of ovigerous females was negatively correlated with depth, which indicates that the low clutch fullness was not induced by fisheries, although the similar phenomenon observed at the Yamato Bank had previously been considered to be due to fishery effects.     

Release size and stocking density for grow-out of <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> in the sea with raft-cultured macroalgae

Sea cucumber, Apostichopus japonicus, is the main cultured species in China. The main culture style for this species is the sea ranching model. Field trials were conducted in bottom-cages to preliminary reveal optimal releasing size, as well as maximum density of A. japonicus in an integrated multi-trophic aquaculture sea ranching area. Different sizes (4–30 g ind^?1) and densities (336–1342 g m^?2) of sea cucumbers were cultured for 13 months in Rongcheng Bay, Shandong Province, China. The size experiment showed that sea cucumber of all sizes grew throughout the experimental period. Sea cucumbers <15 g had high mortality in summer and low SGR in winter, while larger individual (>20 g ind^?1) had no advantage of growth. Sea cucumber sizes of 15–20 g may be suitable for release, considering their higher survival rate and SGR. The density experiment showed that the high biomass group had a low SGR and that the maximum release biomass was 793 g m^?2 based on a regression analysis. The optimal practical release season for sea cucumber was spring based on the results of two field experiments.     

Biomass fluctuation of two dominant lanternfish <Emphasis Type="Italic">Diaphus</Emphasis><Emphasis Type="Italic">garmani</Emphasis> and <Emphasis Type="Italic">D. chrysorhynchus</Emphasis> with environmental changes in the East China Sea

Acoustic surveys have been conducted for estimating the biomass of commercially important fish (e.g., anchovy, jack mackerel), lanternfish (Diaphus garmani and D. chrysorhynchus), and pearlside (Maurolicus japonicus) in summer in the East China Sea (ECS) since 1997. The biomass of lanternfish and pearlside was 2.26–19.16 times that of commercially important fish, and these species represented substantial biomass in the ECS. Though there were no correlations between biomass of pearlside and environmental indices, significant correlations between biomass of lanternfish and southern oscillation index (SOI) in March (positive correlation), arctic oscillation (AO) in March (negative) and October (positive), monsoon index (MOI) in February (positive), and Kuroshio flow mass in winter (positive) were observed. Weak AO and strong MOI would cool down the sea temperature and would lead to increased primary and secondary production in the ECS, thereby enhancing larval survival of lanternfish. The SOI would affect the Kuroshio meander in the ECS, and strong SOI and Kuroshio flow mass would transport larvae of lanternfish to the present survey area. This is the first report on the lanternfish standing stock and its fluctuation in the ECS.     

Occurrence of estuarine and sea eels <Emphasis Type="Italic">Anguilla japonica</Emphasis> and a migrating silver eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis> in the Tokyo Bay area,Japan

Otolith microchemical analyses of the strontium (Sr) and calcium (Ca) concentrations in the eels Anguilla japonica and A. anguilla caught in Tokyo Bay were undertaken to reconstruct the eels’ migratory histories. A. japonica in the yellow stage (immature stage) were caught in a bay without any adjacent rivers or streams. A. anguilla was in the silver stage (early maturing stage), and the eel was confirmed to have just begun spawning migration to the open ocean from Tokyo Bay based on the otolith Sr:Ca ratios, which showed a typical catadromous life history with low Sr:Ca ratio values throughout the eel’s life after recruitment. The mean Sr:Ca ratios in A. japonica from the elver mark to the otolith edge indicated the eels belonged to several general categories of migratory histories, including sea eels (average Sr:Ca ratio ≥6.0 × 10⁻³) and estuarine eels (average Sr:Ca ratio 2.5 to 6.0 × 10⁻³) based on the criteria reported previously in A. japonica. All eels had a certain freshwater life period, although the period was highly variable among fish. These results indicate that A. japonica has a flexible pattern of migration, with the ability to adapt to various habitats and salinities.     

Spatial variation of quantitative color traits in green and black types of sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> (Stichopodidae) using image processing

It has been suggested that the Japanese sea cucumber, Apostichopus japonicus, has three color types (red, green, and black), although the qualitative difference between the color types, particularly between the green and black types, is unclear because of continuous color variation among color types. This study elucidated the color variation between green and black types using image processing (RGB, red–green–blue system) and multivariate analysis to demonstrate whether or not the black and green types can be quantitatively classified. Moreover, spatial variation of the RGB value among various local sites was clarified to estimate potential environmental factors that may affect the color variation. The series of analyses revealed that a quantitative boundary between green and black types could be provisionally established, and also that spatial variability in the intermediate (continuous) color trait between green and black types was significant. Potential environmental factors (depth and industrial activity index) were correlated with the color traits in both color types. These results suggest that the green and black types cannot be regarded as independent color traits and that the color variation between green and black types may be influenced by local environmental factors.