Effects of dietary peptide intake on growth,energy budget,body composition and non‐specific immunity of the sea cucumber Apostichopus japonicus (Selenka)

In this study, a 60‐day feeding trial was conducted to determine the effects of feed supplementating the feed of Apostichopus japonicus with peptides on its growth, energy budget, body composition and immune responses. Sea cucumbers were fed with five experimental diets supplemented with different proportions of peptides: 0 (D1), 12.5 (D2), 25 (D3), 37.5 (D4) and 50 (D5) g/kg in basal diet. Our results suggested that specific growth rate (SGR) and ratio of viscera to body wall (RVBW) of sea cucumbers fed with D4 were significantly improved. Relative to D1, ingestion rate (IR) and faeces production rate (FPR) for D3 and D4 were reduced considerably. Notably, the energy intake increased when peptide level increased from 0 to 50 g/kg. Meanwhile, the energy deposited for growth increased, and the energy loss decreased when peptide level increased from 0 to 37.5 g/kg. The sea cucumbers in D4 had the highest level of crude fat and lowest crude ash. The activities of immunoenzyme, such as SOD, CAT, T‐AOC, ACP and AKP, increased with peptide increase. Results suggested that supplementation of the feed with 25–37.5 g/kg peptides could significantly improve the growth performance, body composition and immune capacity of Apostichopus japonicus.     

Requirement of vitamin E of growing sea cucumber Apostichopus japonicus Selenka

This feeding trial was conducted to determine the vitamin E requirement of growing sea cucumber Apostichopus japonicus Selenka. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded levels of vitamin E (6.7, 81.2, 159.3, 237.8, 314.6, 395.9 mg/kg diet). Each diet was assigned randomly to 30 growing sea cucumber with initial body weight 15.43 g in triplicates for 8 weeks. Survival rate was not affected by dietary vitamin E; meanwhile, both the weight gain (WG) and specific growth rate were presented the trend of increasing first and then kept stable. With the increasing of dietary vitamin E, crude lipid content of body wall was evaluated firstly and dropped afterwards. When dietary vitamin E contents were lower than 159.3 mg/kg, vitamin E contents of body wall were increased by dietary vitamin E levels, but there were no more differences when dietary vitamin E higher than 159.3 mg/kg. There were minor effects on digestive enzymes of intestine by dietary vitamin E. Contents of malondialdehyde were decreased, while the total superoxide dismutase activity was increased first and then decreased with the increasing of dietary vitamin E. Activities of intestinal alkaline phosphatase were increased, and glutamic oxaloacetic transaminase were decreased first and then increased with the increasing of dietary vitamin E. In conclusion, analysis by a linear regression equation of WG or vitamin E contents in body wall indicated that the optimum requirement of vitamin E for growing sea cucumber (initial body weight 15.43 g) was 165.2–187.2 mg/kg diet.     

Effects of l‐tryptophan on the performance,energy partitioning and endocrine response of Japanese sea cucumber (Apostichopus japonicus Selenka) exposed to crowding stress

The effects of dietary tryptophan on growth performance, energy budget and endocrine response of sea cucumber Apostichopus japonicus (Selenka) were studied, to test whether the tryptophan could mitigate the crowding stress response of the sea cucumber. Four density treatments of the sea cucumber (i.e. 4, 8, 16 and 32 individuals per 40 L water, represented as L, ML, MH and H) were fed with diets containing 0 (control), 1%, 3% and 5% l ‐tryptophan, respectively, for 75 days. The results showed that the specific growth rate (SGR) and feed conversion ratio (FCE) of A. japonicus decreased with increasing in the stocking densities but increased after supplementation of 1% and 3% tryptophan. The energy allocation of the A. japonicus was affected significantly by stocking density and dietary tryptophan treatments (p < .05). In general, the energy consumed (C) and energy used for growth (G) decreased with the increasing in the stocking densities. The highest proportion of energy deposited to growth occurred in each 3% treatment (L, 6.38%; ML, 6.12%; MH, 4.56% and H, 4.14%), which were significantly higher than the controls (p < .05) respectively. Higher cortisol, glucose and lactate levels of A. japonicus were recorded in the control treatments of MH and H, but l ‐tryptophan supplementation could reduce the levels of cortisol, glucose and lactate of the test animals. The present results proved that the growth performance of the sea cucumber was inhibited by high stocking densities, while dietary supplementation of tryptophan could significantly mitigate the crowding stress, and improve the growth of sea cucumber.     

Effect of salinity on growth and energy budget of red and green colour variant sea cucumber Apostichopus japonicus (Selenca)

Sea cucumber, Apostichopus japonicus (Selenca), tolerates salinity fluctuations inhabiting intertide zone. This study deals with growth, food intake, food conversion and the bioenergetic responses of the red variant (wet weight of 2.60 ± 0.11g) and green variant (wet weight of 2.56 ± 0.08 g) A. japonicus to different salinities of 22, 26, 30, 34, and 38 psu at 16.5 ± 0.5°C. The results showed that salinity had a significant effect on specific growth rate (SGR) of both green and red variants A. japonicus (P < 0.05). Both colour variants of sea cucumber had highest SGR at 30 psu, and then decreased when salinity below or above this point. Maximum SGR (the green 1.07 ± 0.08% day^?1, the red 1.14 ± 0.09% day^?1 respectively) is related with maximum food intake (FI) and highest food conversion efficiency (FCE) (P < 0.05) occurring at 30 psu. Only under 22 psu, the green variant grew faster than the red variant (P < 0.05), and under other four salinity treatments there was no significant difference between SGR of two colour variant holothruians (P > 0.05). Values of adaptable salinity scope for green and red variants sea cucumber survival are 18.5~39 psu and 20.9~38.6 psu respectively. The average energy budget formula of sea cucumber at 30 psu was: 100C = 6G +42F +3U+49R (C, energy ingested; G, energy for growth; F, energy loss as faeces; U, energy used for ammonia excretion; R, energy loss for respiration). The sea cucumber had maximum energy ingested (C) and highest proportion of energy for growth (G) at 30 psu, and then decreased when salinity is above or below this salinity. Both red and green variants of A. japonicus deposited for growth were very low, and the energy loss in faeces and energy for respiration accounted for the majority of assimilation energy. The result clearly showed that the optimum condition for farming green and red variants A. japonicus, both with respect growth and energy allocation, is the salinity scope of 26 ~ 30 psu.     

Utilization of dietary carbohydrates by sea cucumber Apostichopus japonicus (Selenka) as indicated by carbon stable isotope analysis

Carbon stable isotopes were used as trophic markers to investigate the utilization of dietary terrestrial‐source carbohydrates by sea cucumber Apostichopus japonicus. Sea cucumbers were fed by five different types of diets with the ingredients containing Sargassum muticum either without starch or with one of the four starches including corn starch, sweet potato starch, tapioca starch and potato starch. After the 70‐day feeding trial, the carbon isotopic compositions of A. japonicus appeared to reflect those of corresponding dietary components. The average contribution of corn starch (22.0%) to the growth of A. japonicus was slightly higher than the expected contribution (20%). While the proportional contributions of sweet potato, tapioca and potato starches (6.0%, 7.0% and 4.0%, respectively) were all considerably lower relative to the expected contributions. These results indicated that A. japonicus could utilize corn starch more efficiently than sweet potato, tapioca or potato starch. Moreover, A. japonicus fed diet containing corn starch showed the highest specific growth rates which were significantly higher than those fed diets containing potato or tapioca starch. The results of the present study suggested that the corn starch could be used as dietary carbohydrate source in the artificial feeds for A. japonicus farming.     

饲料中不同碳水化合物水平对刺参幼参生长和能量收支的影响

通过设置不同碳水化合物水平(25.61%,34.55%,45.31%,55.82%,66.09%),探讨对刺参幼参[体重(1.67±0.06)g]生长、饲料利用、体组成和能量收支的影响。实验周期为60 d,每组设3个重复,每个重复40头幼参。结果表明:(1)随着摄食碳水化合物水平的升高,特定生长率和饲料转化率均呈现先上升后下降的趋势。基于SGR和FCE的二次回归模型估算幼参碳水化合物的最适需求量为50.15%~53.95%;(2)蛋白质效率和摄食能在45.31%~66.09%碳水化合物水平组均显著高于25.61%~34.55%组,说明碳水化合物为幼参获得蛋白质节约效应提供了能量;(3)饲料中碳水化合物水平对幼参摄食率、排粪率和表观消化率均有显著性影响,适宜的碳水化合物水平会提高机体对营养物质的利用效率;(4)全参、体壁及消化道中粗脂肪含量均随摄食碳水化合物水平的升高而有所增加,表明摄食过量碳水化合物会造成幼参体组织脂肪的沉积;(5)在66.09%碳水化合物水平下,幼参生长能占摄食能的比例显著降低,粪能、排泄能及呼吸能占摄食能的比例有所升高,说明过高的碳水化合物水平会造成机体能量利用效率的降低。综上,适宜的碳水化合物水平可以促进幼参生长,提高饲料利用效率,而摄食过量碳水化合物会造成其体组织脂肪的沉积,并影响机体能量代谢水平。     

Effect of replacing dietary fish meal with silkworm (Bombyx mori L) caterpillar meal on growth and non‐specific immunity of sea cucumber Apostichopus japonicus (Selenka)

The effect of dietary substitution of silkworm (Bombyx mori L) meal (SM) for fishmeal (FM) on the growth performance and non‐specific immunity of sea cucumber (Apostichopus japonicus) (initial weight: 12.8 ± 0.16 g) was determined. Four isonitrogenous and isocaloric diets were formulated: Diet 1, which served as the control diet, contained 5% FM; Diet 2 contained 3.75% FM and 1.25% SM; Diet 3 contained 2.5% FM and 2.5% SM; and Diet 4 contained 5% SM. Other ingredients in each of the four diets were kept in the same proportion. After 8 weeks of feeding, the results showed that sea cucumbers fed Diet 2 had 18.7% increases in weight over those fed the control diet, but no significant difference was observed. No obvious difference in body wall composition was detected among the sea cucumbers fed the four different diets. Immunity analysis indicated that phagocytosis and serum alkaline phosphatase activity were not significantly (P > 0.05) affected when FM was partially or completely replaced with SM. Serum lysozyme activity of sea cucumbers fed Diet 4 showed a significant (P < 0.05) growth increase compared with those fed control diet. The results revealed that SM could be an effective substitute for FM in sea cucumber diet.     

Cryopreservation of sea cucumber Apostichopus japonicus (Selenka) sperm

A simple and convenient method for the cryopreservation of sea cucumber Apostichopus japonicus (Selenka) sperm was tested in the present study. The highest motility (76.7±2.9%) of post‐thawing sperm was obtained in 15% dimethyl sulphoxide (DMSO) with a 1:9 dilution (semen volume to DMSO volume) when 0.5 mL semen–DMSO mixture was frozen at 6 cm above liquid N₂ in a closed styrofoam box. After thawing, sperm cryopreserved in glycerol almost lost motility entirely. Although there was no significant difference in percentage of motile sperm between 15% and 20% DMSO, the duration of sperm motility of 15% DMSO group was longer than that of 20% DMSO group. The motility of post‐thawing sperm enhanced when the dilution ratio of semen increased from 1:1 to 1:9. Morphological changes such as the loss of mitochondria, swollen plasma membrane and broken or rolled‐up tails were observed in post‐thawing sperm using an eosin–nigrosin staining. The fertility of cryopreserved sperm was significantly lower than that of unfrozen sperm. The 10‐fold increase in sperm to egg ratio resulted in double fertility for cryopreserved sperm, and about 70% fertility relative to the control.     

Effects of stocking density and body physical contact on growth of sea cucumber,Apostichopus japonicus

The present study was conducted to investigate the effects of physical contact on the growth of Apostichopus japonicus through various stocking densities, group culture and physical isolation culture. The results showed that the specific growth rates (SGR), the feed conversion efficiencies and the energy used for growth of the animal decreased with the increase in stocking density, while the coefficient of variations (CV) in group culture treatments increased. In high density treatments (4, 6 ind./40 L), the SGRs in isolated culture treatments were not different to those in group culture treatments of the same density, however, the CVs in the latter (58.0% and 60.4%, respectively) were significantly higher than those in the former (37.8% and 35.8%, respectively). The CV difference between group one and isolated one of the same density (20.2% and 24.7%, respectively) was less than that in single individual culture (32.7%), indicating that the effect of physical contact on growth variation was weaker than that of genetic factor. This study proved that the individual growth variation in sea cucumber resulted mainly from genetic factor and physical contact, while some chemical mediator might also be the factor affecting the growth rate of sea cucumber.     

An annual faecal 16S amplicon sequencing of individual sea cucumber (Apostichopus japonicus) demonstrates the feeding behaviours against eukaryotes in natural environments

To fill in the gaps in knowledge as to how individual sea cucumber Apostichopus japonicus behave consuming eukaryotic food sources in natural environments, eukaryotic communities in the faeces of sea cucumbers and sediments were analysed through one whole year based on 16S rRNA gene sequencing of the organelle genomes. A total of 390 eukaryotic features were obtained, and 99.7% of the features were assigned to chloroplasts. The eukaryotic communities in faeces and sediments showed seasonal fluctuations through one whole year based on Bray‐Curtis distance and community composition. Comparison of eukaryotic communities between faeces and sediments showed that 12 families including Chaetocerotaceae and Laminariaceae were more abundant in faeces than in sediments, suggesting that sea cucumbers may choose sediment containing these algal taxa more often compared with others in natural environments. All features of Laminariaceae were assigned to Saccharina japonica, which is consistent with the fact that this alga is one of the most suitable diets in the aquaculture of A. japonicus. Assessments of individual 16S amplicon sequences of both faecal and sediment samples could be an alternative tool to help us understand dynamic feeding behaviours of sea cucumber populations in contributing to bioresource conservation and development of a superior approach to aquaculture.     

Effect of temperature on growth,digestion and immunity of green,white and purple sea cucumbers,Apostichopus japonicus (Selenka)

Three color morphs (white, green and purple strains) of Apostichopus japonicus (Selenka) were cultured in artificial seawater, for approximately 90 days, in three temperature ranges: 27–22°C (high), 22–17°C (mid) and 17–12°C (low). All strains grew in all temperature ranges. Temperature significantly affected growth rate, digestibility, digestive enzymes and immune‐related enzymes. Highest specific growth rates were exhibited in 4‐month‐old sea cucumbers at mid and high temperatures, and in 16‐month‐old sea cucumbers at mid and low temperatures. Specific growth rates of green and purple strains were not significantly different, but were significantly higher than that of the white strain at mid temperatures. The digestibility of each strain was significantly higher at 27°C, 22°C and 17°C than at 12°C. Green‐strain digestibility was higher than that of purple and white strains at specific temperatures. Protease and amylase activities of all strains followed bell‐shaped temperature curves with maximum digestive enzyme activity at 17°C. The activities of alkaline and acid phosphatases were higher in the guts of the green strain than in the white or purple strains at the same temperature. Superoxide dismutase activity was higher in the purple strain than in white and green strains.     

Effects of light intensity on daily activity rhythm of juvenile sea cucumber,Apostichopus japonicus (Selenka)

Eight light‐intensity treatments (natural light, continuous darkness, and 15, 30, 60, 125, 250 and 500 lx under LD 12:12 cycle) were used to investigate the effects of light intensity on the daily activity of 30.27±3.08 g sea cucumber Apostichopus japonicus (Selenka). Cyclic nocturnal activity patterns of behaviour were observed at different light intensities in the range of 15–500 lx under LD 12:12 cycle. And an ongoing nocturnal cycle persisted in DD cycle for up to 8 days, but longer feeding time and less marked rhythm occurred at continuous darkness. Under poor light conditions (I<5.18 lx), the daily activity rhythm of A. japonicus was governed by an innate biological clock and the effect of light intensity was not significant among different treatments. And more individuals tended to retreat to shelters (from 56.04% to 91.83%) with the increase of light intensity within the weak light condition (from 5.18 to 278 lx). However, the daily behaviours of A. japonicus were influenced under strong light conditions (>278 lx). Less than 8.17% individuals kept actively feeding and the proportion was not decreased with the increase of light intensity.     

Fatty acid composition in sea cucumber Apostichopus japonicus fed with microbially degraded dietary sources

Fatty acids (FAs) in the body walls of sea cucumber, Apostichopus japonicus, reared under diet‐administered conditions were analysed to determine the accumulative status of dietary FAs. Four test diets different in diet source and microbial degradation, that is, a diet composed of powdered pennate diatoms (diet PD), one microbially degraded of diet PD (diet PDM), one composed of brown algae [tangle weed, Saccharina (Laminaria) longissima (Miyabe); diet BA], and one microbially degraded of diet BA (diet BAM), were fed to juvenile A. japonicus, respectively, for 2 months and the FA composition of the juvenile body wall was determined. FAs in body wall were influenced and changed by diet source and microbial degradation of diet. Degradation significantly promoted juvenile's growth and increased iso‐15:0 in the body wall; however, iso‐15:0 was not a growth factor. FA 20:4n6 and 20:5n3 were dominant in all groups despite being a minor dietary constituent, suggesting that these FAs can be readily accumulated or synthesized in the animal.     

Effects of dietary macroalgae meal and lipid source on growth performance and body wall fatty acid composition of sea cucumber Apostichopus japonicus

A 70‐day experiment was conducted to examine the effects of different macroalgal meals and lipid sources on growth, body wall composition and fatty acid (FA) profile of sea cucumber Apostichopus japonicus. Two macroalgal meals including Sargassum muticum (SM) and Gracilaria lemaneiformis (GL) and two lipid sources including fish oil (FO) and vegetable oil (VO) were formulated into four diets, i.e., S. muticum and fish oil (SF), S. muticum and vegetable oil (SV), G. lemaneiformis and fish oil (GF) and G. lemaneiformis and vegetable oil (GV). The results showed that the specific growth rates (SGR) of A. japonicus fed diets containing SM were significantly higher than those fed diets containing GL. No significant differences in SGR between the FO‐based and VO‐based groups were observed. Similar results were observed in the body wall lipid content. Most body wall FAs changed to resemble the dietary FA proportions because of the dietary effect. Concentrations of 20:4n‐6 of the SF and GF groups were significantly lower than the SV and GV groups, while levels of 20:5n‐3 and 22:6n‐3 were significantly higher than the SV and GV groups. The n‐3/n‐6 polyunsaturated fatty acids (PUFA) ratios of the SF and GF groups were significantly higher than the SV and GV groups. Moreover, the SF group had significantly higher 20:5n‐3 and 22:6n‐3 contents and n‐3/n‐6 PUFAs ratio than the GF group. These findings reveal that the SF diet can show beneficial effects on both growth performance and body wall n‐3 PUFAs content of A. japonicus.     

An artificial oyster‐shell reef for the culture and stock enhancement of sea cucumber,Apostichopus japonicus,in shallow seawater

Because of its high nutritional value and health benefits, aquaculture production of Apostichopus japonicus in China is the largest of any single species. Therefore, the development of new farming methods is of considerable significance. In this study, discarded oyster shells have been used to create an artificial reef for the culture of this species. The results have shown that from 6th March 2009 to 26th November 2009, the wet weight of sea cucumber increased from 49.57 ± 1.16 to 79.87 ± 1.46 g ind^?1. Between 16th July and 18th October, the specific growth rate and daily weight gain of A. japonicus differed significantly from other periods. Population density was higher within the reef compared with outside the reef area, and the difference was significant (P < 0.01). The maximum distance between A. japonicus individuals within the reef area on 3rd March and 16th July was 65.0 ± 3.3 and 62.9 ± 4.4 cm, respectively, but the difference was not significant (P > 0.05). The diatom species attached to the oyster shells were similar to those found in the stomach content of A. japonicus. In conclusion, the oyster‐shell reef provides a suitable habitat and shelter for the culture of the sea cucumber, A. japonicus.     

Microbial communities in sea cucumber (Apostichopus japonicus) culture pond and the effects of environmental factors

This study investigated microbial community composition as well as their correlation with environmental factors of Apostichopus japonicus culture ponds in northern China by 16S rRNA gene amplicon sequencing. The results showed that microbiota richness varied consistently with diversity in the pond ecosystem. Microbiota richness and diversity were highest in sediment, followed by gut of A. japonicus and water. The dominant bacterial phylum in the pond ecosystem is Proteobacteria. Gammaproteobacteriaeria and Flavobacteria are two dominant bacterial classes in the ecosystem. There is significant difference (p < 0.05) between dominant bacterial communities at the levels of order, family and genus. There is also remarkable regional difference (p < 0.05) between microbial community composition in the pond ecosystems. Specifically, microbial community composition in Changhai and Yingkou show a high similarity, so do those of Laoting and Rushan. According to the redundancy analysis of the microbial community composition and pond environmental factors, chemical oxygen demand is the dominant environmental factor determining microbial community composition in pond water; sulphide has the greatest influence on the microbial community composition in pond sediment; the rest of environmental factors have varied influence on microbial community composition in pond ecosystems.     

Spontaneous captive breeding and larval development in the green and red variants of the Japanese sea cucumber Apostichopus japonicus (Selenka 1867)

We report the spontaneous spawning, larval development, survival rate and larval growth rate patterns observed in the green and red variants of the Japanese sea cucumber Apostichopus japonicus. The green variant adapted well to the captive conditions in the Sasebo City Fisheries Center and spontaneously spawned without any induction or stimulation. One hundred individual green variants spawned nine times and produced approximately 155 million eggs. In contrast, 50 individual red variants showed poor adaptation to captivity and spawned spontaneously only three times, producing about 12 million eggs. Larval development and growth rate pattern was almost identical between the two variants of A. japonicus. In contrast, the larval survival rate for the green variant was over 90% up to the auricularia stage (10 days), but much low (less than 30%) for the red variant. We demonstrated that the green variant of A. japonicus was easier to rear in captivity. This provides a useful method for maintaining sustainable harvests and eventually contributing to restocking and sea ranching of the existing population. Further studies about optimal ecological conditions and behaviour are needed to improve egg production and survival rate of the red variant of A. japonicus.     

Energy budget adjustment of sea cucumber Apostichopus japonicus during breeding period

Reproductive success is unpredictable for many cultured echinoderms. And cost of reproduction plays an important role in animal life‐history. Therefore, understanding cost of reproduction contributes to improving breeding techniques during broodstock rearing in aquaculture. In this study, energetic costs during breeding were explored in the sea cucumber, Apostichopus japonicus, an important aquaculture species in China, Japan and South Korea. The food intake, digestive functions and energy budget patterns were estimated at different breeding stages. Unexpectedly, in the growing gonad phase of the sea cucumber lifecycle, animals showed a decline in feed consumption (from 4.49 to 2.6 g ind^?1 day^?1), ingestion rate (from 0.4 to 0.13 g g^?1 day^?1) and apparent digestive rate (from 14.73% to 10.92%), resulting in a reduced energy input. In addition, the increased energy investment in reproduction (from 1.76% to 15.61%) resulted in a decrease in energy allocated to soma growth (from 17.1% to 5.64%) and self‐maintenance (from 47.82% to 34.67%). These results suggest that reproduction impairs energy acquisition ability in breeding A. japonicus. And broodstock can adapt energy shortage by internal adjustment of energy allocation strategy.