Optimum dietary protein level for growth and coelomic fluid non‐specific immune enzymes of sea cucumber Apostichopus japonicus juvenile

A feeding trial was conducted to determine the optimum dietary protein level of sea cucumber Apostichopus japonicus juvenile focusing on growth performance and non‐specific immune response. Diets with seven crude protein levels (42.0, 108.9, 155.2, 216.7, 258.0, 313.3 and 357.5 g kg^?1) were fed to sea cucumber juveniles (1.05 ±0.01 g) once a day for 100 days. More than 70% survival was observed, and there was no significant difference among all treatments. The sea cucumbers fed diets containing 108.9 g kg^?1 crude protein showed significantly (P < 0.05) higher body weight gain than those of the sea cucumbers fed diets containing 42.0, 216.7, 258.0, 313.3 and 357.5 g kg^?1 crude protein. No significantly differences (P > 0.05) were observed in moisture, crude protein, crude lipid, ash and carbohydrate content of the body wall among all treatments. The coelomic fluid catalase activity of the sea cucumbers generally increased with increasing dietary protein levels. Therefore, the acid phosphatase, superoxide dismutase and lysozyme activity increased with increasing dietary protein levels at first and decreased subsequently. The relationship between dietary protein levels and body weight gain was analysed by a second‐order polynomial regression analysis model. The result indicates that the optimum dietary protein level for sea cucumber juveniles is 135.4 g kg^?1.     

不同光照强度对刺参幼参生长的影响

采用室内受控的实验方法研究了不同光照强度对刺参幼参生长的影响。实验设暗光、微强光、强光3个光照处理,同时每个光照处理组分别投喂两种饵料(人工配合饲料、人工配合饲料加底栖硅藻)。研究结果表明,不同光照强度下,幼参的生长差异显著(P<0.05),每种饵料处理下强光处理组幼参的特定生长率和日增重明显高于暗光处理组,而相同光照强度下两种饵料处理对幼参生长的影响差异不显著(P>0.05)。3个光照处理下幼参的特定生长率平均分别为0.26%、0.63%、0.98%/d,日增重为0.53、1.39、2.26 g/d。     

Effect of dietary inclusion of various plant ingredients as an alternative for Sargassum thunbergii on growth and body composition of juvenile sea cucumber Apostichopus japonicus

Two feeding experiments were conducted to investigate the effects of dietary inclusion of different plant ingredients on growth and body composition of juvenile sea cucumber. In the first experiment, three replicate groups of the sea cucumber (average weight of 1.5 ± 0.05 g) were fed one of the seven experimental diets containing Sargassum thunbergii (CON), Undaria (UND), Laminaria (LAM), Brassica oleracea L. var. capitata (BRA), fermented soybean (FSB), distillers dried grain (DDG) and rice straw powder (RIC) for 10 weeks. After the feeding trial, survival was not significantly different among the dietary treatments. Specific growth rate (SGR) of sea cucumber fed the FSB diet (0.73) was significantly higher than that of sea cucumber fed the other diets (P < 0.05), except for the DDG diet (0.64). The lowest SGR was observed in sea cucumber fed the CON diet (0.14). Based on the result of the first experiment, the second experiment was conducted to determine the proper level of dietary DDG for growth of juvenile sea cucumber. Three replicate groups of sea cucumber (average weight of 1.6 ± 0.02 g) were fed one of the five experimental diets containing different levels (g kg^?1) of DDG (DDG0, DDG10, DDG20, DDG30 and DDG40) for 12 weeks. Survival was not significantly different among the treatments (P > 0.05). The best SGR was obtained in sea cucumber fed the DDG10 diet (10.1) and the value of sea cucumber fed the DDG40 diet (5.6) was the lowest (P < 0.05). The SGR of sea cucumber fed the DDG10 diet did not differ from that of sea cucumber fed the DDG20 diet (P > 0.05). Moisture, crude protein, crude lipid and ash contents of whole body in sea cucumber among the dietary treatments (P > 0.05) did not differ significantly at both experiments. Results of two feeding trial suggest that dietary inclusion with 100 g kg^?1 FSB or 100–200 g kg^?1 DDG may improve growth of juvenile sea cucumber, and especially DDG could be used as a good ingredient for the low‐cost feed formulation.     

Effects of dietary lipid level on growth performance,body composition and digestive enzymes activity of juvenile sea cucumber,Apostichopus japonicus

A feeding trial aimed to determine the effects of dietary lipid level on growth performance, body composition and digestive enzymes activity of juvenile sea cucumber, Apostichopus japonicus. Diets with six crude lipid levels (1.9, 13.8, 29.1, 43.6, 59.6 and 71.6 g kg^?1) were fed to sea cucumbers (initial weights 0.65 ± 0.01 g) at a density of 30 juveniles, once a day. After 60 days, body weight gain (BWG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) decreased with increasing dietary lipid levels. The sea cucumbers fed 1.9 g kg^?1 crude lipid showed significantly higher (P < 0.05) BWG than those of the sea cucumbers fed 59.6 and 71.6 g kg^?1 crude lipid. Intestinal protease and lipase activities generally increased with increasing dietary lipid levels. Eicosapentaenoic acid (EPA) content of body walls generally increased with increasing dietary lipid levels. Docosahexaenoic acid (DHA) content of body walls reached the maximum value at a dietary lipid level of 13.8 g kg^?1. N‐3 highly unsaturated fatty acid content followed the same pattern of DHA. According to the growth performance and body composition of sea cucumbers, it can be indicated that the optimum dietary lipid level for juvenile sea cucumbers is between 1.9 and 13.8 g kg^?1.     

饲料中不同脂肪酸搭配对仿刺参幼参生长和体组分的影响

在水温13．0～23．0℃下,将平均质量1.45g的仿刺参Apostichopus japonicus幼参放养在容量50L（45cm×31cm×30cm）的水槽中,投喂以鱼粉、虾糠和玉米蛋白为蛋白源,添加马尾藻粉、海泥、贝壳粉和黄原胶等粗蛋白12．70％和粗脂肪5．48％的饲料。在饲料中含有不同比例的亚油酸（LA）、α-亚麻酸（LNA）、二十二碳六烯酸（22：6n-3）（DHA）和二十碳五烯酸（20：5n-3）（EPA）,以橄榄油平衡脂肪含量,以贝壳粉平衡其他成分,配制成LA和LNA的比例分别为20：0（S1）;20：1（S2）;1：2（S3）和2：1（S4）;LA、LNA、DHA和EPA的比例分别为6：1：5：0（S5）;10：1：30：5（S6）和10：5：6：1（S7）的膏状饲料。80d的饲养表明,幼参摄食只添加橄榄油（S1）、添加橄榄油和亚油酸（S2）的饲料时生长最慢,显著低于其他各组的幼参（P〈0．05）;摄食只添加亚麻酸（S3）或亚油酸和亚麻酸（S4）的饲料时,生长较慢,二者差异不显著（P〉0．05）,但均显著低于摄食添加DHA（S5）和EPA＋DHA（S6）的幼参（P〈0．05）;摄食添加上述4种脂肪酸（S7）饲料时,生长最快,即LA,LNA,DHA和EPA的比例为10：5：6：1时,体壁最厚,肠道中淀粉酶和蛋白酶的有效活力最高,对饲料脂肪和蛋白质的消化率显著高于其他各组（P〈0．05）。摄食实验饲料的仿刺参体内蛋白＋脂肪十灰分的含量之和（平均85．79％）远大于初始刺参（79．70％）的相应之和,由高至低依次为：S2（90．57％）〉S1（87．97％）〉S3（85．78％）〉S4（85．29％）〉S7（84．19％）〉S5（83．79％）〉S6（82．97％）。仿刺参摄食后体内18碳脂肪酸含量呈负增长,而体内EPA的含量均呈正增长,且恒定变化在4．36％～6．83％之间,表明饲料中的18碳脂肪酸能转化为仿刺参体内的EPA。添加了4种脂肪酸的S4、S5、S6和S7组幼参肠过氧化氢?     

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.     

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.     

Influence of vibration caused by sound on migration of sea cucumber Apostichopus japonicus

Vibrations exist widely in the ocean, and the one caused by sound and biological movement plays an important role in the sensory system of marine organisms. The vibration caused by sound has many characteristics that make it a good candidate for modifying the movements of marine organisms. However, few studies have tested whether hydraulic vibration caused by sound has effects on the migration of sea cucumbers, and this has never been systematically studied previously in Apostichopus japonicus (Selenka). In order to understand whether hydraulic vibration caused by sound at different frequencies could affect the migration of sea cucumbers, in the present study, hydraulic vibration caused by sound at various frequencies was tested in the laboratory to determine their effects on the migration of A. japonicus at different sizes. The mean probability distribution was used as a statistical index to demonstrate the moving tendencies of the species. The experimental results showed that medium and small A. japonicus (<10 g ind.^?1) tend to move towards the low‐frequency vibration source caused by sound (100 Hz), whereas all sizes of A. japonicus tend to move backwards the high‐frequency vibration source caused by sound (10 000 and 28 000 Hz). This study provides basic data on the behavioural ecology of this species, as well as theoretical reference for the design of artificial reefs and harvesting equipment for A. japonicus.     

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.     

间歇投喂添加蛋氨酸硒饲料对仿刺参（Apostichopus japonicus Selenka）幼参生长和免疫指标的影响

在水温9℃～13℃下,将平均体质量0.745±0.3 g的仿刺参（Apostichopus japonicus Selenka）随机投放到50cm×40cm×30cm的塑料水槽内,每箱10头,共分为五组,每组3个平行,采用不同间隔时间投喂每kg添加0.5 mg蛋氨酸硒的饲料：第1、2、3、4组分别间隔0、5、10和15...     

饲料中适宜的蛋白质和氨基酸水平对刺参生长的影响

利用单因素设计,以初始体重为4.5～4.8g的刺参Apostichopus japonicus为研究对象,分别配制以鱼粉、马尾藻和酒糟等为主要成分的配合饲料(蛋白含量为14.7%、17.7%、19.1%及21.5%)喂养刺参70d。实验表明,实验刺参的增重率和对配合饲料的消化率随着饲料中蛋白含量的增加而增加。配合饲料中粗蛋白的含量为21.49%时,试验幼参的增重率及蛋白质消化率达到最大。另一组试验对5种含有不同种类和数量的氨基酸配合饲料进行了饲喂对比试验,实验进行了40d。结果表明,饲喂富含甘氨酸、蛋氨酸、赖氨酸和精氨酸饲料的刺参增重率最高。     

The concentrating method of benthic diatom affects the growth of juvenile sea cucumber (Apostichopus japonicus) and water quality

Concentrating methods generally affect the vitality of the microalgae and may alter its feeding and environmental effect. This article studied the concentrating methods (centrifugation and settlement) on primary productivity of the microalgae Cylindrotheca fusiformis. The effects of free settled C. fusiformis, centrifuged C. fusiformis and macroalgae Sargassum polycystum on growth of the sea cucumber, farming water and sediment quality under two light intensities (1500 lx and 500 lx) were studied by a six‐week rearing experiment. The results showed that the primary productivity of free settled C. fusiformis was 3–4 times higher than that of those centrifuged, indicating centrifugation inhibited the photosynthesis process. In the rearing experiment, feed, light intensities and their interactions significantly affected the specific growth rate (SGR) of the animals (P < 0.01). The animals reared in higher light intensity also had higher SGR, but only those fed free settled C. fusiformis reached a significant level (P < 0.05). The group also had the highest SGR (P < 0.05), suggesting that the photosynthesis ability of C. fusiformis directly contributed to the growth of the animals. During the experiment, the soluble inorganic nitrogen, reactive phosphorus in farming water, total nitrogen and total organic carbon in the sediment gradually increased in all treatments. The S. polycystum group had slightly higher level of nutrients in water and sediment at the end of the experiment for its high organic content. These results provide evidence that the harvesting methods of C. fusiformis directly affect its vitality, which consequentially alter the feeding and environmental effect, especially under higher light densities.     

Effects of light intensity on larval development and juvenile growth of sea cucumber Apostichopus japonicus

The growth rate, survival rate, development and setting rate of larval sea cucumber Apostichopus japonicus were measured under four light intensities (0, 50, 500 and 2,000 lx), and the growth rate and metabolism of the juvenile sea cucumbers were investigated under four light intensity treatments (0, 100, 1,000 and 2,500 lx). The light requirements (i.e. intensity) of the sea cucumber changed as they grew. Better growth performance, development and survival rate were observed in embryos and larvae under 500 lx treatment. However, reduced light intensity (50 lx) increased the settlement rate of the larvae. The highest specific growth rate occurred in juvenile sea cucumbers under 1,000 lx. The sea cucumbers in this group also had the lowest oxygen consumption rate and ammonia excretion rate, but the highest O:N ratio, indicating that the optimal light intensity for the juvenile sea cucumber culture was 1,000 lx. Therefore, appropriate light intensities were suggested to provide larval and juvenile sea cucumbers with better growth and development conditions.     

A new system for the culture and stock enhancement of sea cucumber,Apostichopus japonicus (Selenka),in cofferdams

A new multilayer, plate‐type system for the culture and stock enhancement of sea cucumbers in cofferdam was developed. To optimize and evaluate the system, four experimental designs were implemented using polyethylene (PE)‐corrugated sheets of various colours, interval spacing and shapes/styles. Results showed that a system equipped with black PE‐corrugated sheets attracted more animals than either blue, green, transparent or a selection of mixed sheets (six transparent sheets in the upper layer and five black sheets in the lower layer) (P<0.05). Also, more animals gathered in the system with oblique‐angled sheets (30° to the base plate) than either a wavy (the bottom and every second sheet was at an angle of 10° to the base plate) or parallel arrangement (P<0.05), and more animals assembled in the system with 2 cm between sheets than spacings of 3, 4 or 5 cm (P<0.05). As expected, the upper layers of the systems attracted more animals than lower layers in most cases except for those with transparent and mixed oblique‐angled sheets with a 3 cm spacing (P<0.05). Thus, a system with black, oblique‐angled‐corrugated sheets and 2 cm spacing is recommended for Apostichopus japonicus (Selenka) culture and stock enhancement in cofferdams or ponds.     

Comparative study of water environment variation in the industrial aquaculture system of sea cucumber Apostichopus japonicus

This study investigated the microbial community, water quality, bacterial densities and growth performance in the industrial aquaculture system of sea cucumber Apostichopus japonicus without water exchange. Six treatments were set including, C (control treatment without sea cucumber culturing), S (small individual treatment), B (big individual treatment), Sd (small individual treatment with high density), Ss (small individual treatment adding carbohydrate source) and Sb (small individual treatment adding effective microorganisms). A total of 27916–32236 optimized reads and 564–742 operational taxonomic units (OTUs) were obtained from each samples. The phylum Proteobacteria, Actinobacteria, Bacteroidetes and Cyanobacteria predominated, representing 69.01–97.21% of the bacterial communities in the water samples. Cluster analysis and principal component analysis (PCA) showed that higher similarity was observed among S, Sd, Ss and Sb. The densities of TB and Vibrio in Ss were significantly higher than those in the other culture treatments after the 7th day. The concentrations of NH₄‐N, NO₂‐N, NO₃‐N and PO₄‐P in Ss and Sb were relatively lower than the other treatments. Conclusively, no deterioration was found in the water environment parameters during the 21‐day culture period without water exchange, indicating that low‐level water exchange protocol may be applied to the industrial aquaculture system. Based on the effects of different operations on culture system, industrial aquaculture is proved to be a viable way to rear sea cucumber.     

Intra‐specific effects of sea cucumber (Apostichopus japonicus) with reference to stocking density and body size

The present studies deal with the intra‐specific effects of sea cucumber Apostichopus japonicus with unlimited food resources, especially the effects of stocking density on growth variation of the animal and energetic changes of small individuals under the stress of large individuals. The results showed that with the initial body weight of 5.12±0.09–6.11±0.26 g of sea cucumber among the densities of 5, 10, 20, 30, 40 and 50 ind./100 L, the density of 20 ind./100 L was the optimum stocking density because of its highest specific growth rate, crude protein content and crude lipid content in tissue. Individual growth variation of A. japonicus increased with the increase of stocking densities, whereas no significant differences in variation were found when the density was over 30 ind./100 L (P>0.05). The low‐weight individuals under the stress of heavy‐weight individuals exhibited obvious changes in energetics, such as lower ingestion rate, lower energy deposited as growth but higher respiration and excretion. The coefficient of variation in growth of the animals was over 70% due to the simultaneous action of aggression and maybe a factor of chemical mediator, and led to significant changes in the energetics of small‐sized individuals.     

Microsatellite genetic variation in wild and hatchery populations of the sea cucumber (Apostichopus japonicus Selenka) from northern China

Farming of the sea cucumber Apostichopus japonicus (Selenka) started 20 years ago and is still in rapid expansion in China. In order to assess the genetic status of both wild and cultivated stocks of this species, we used eight microsatellite markers to estimate the level of genetic diversity within five hatchery stocks and two wild populations of A. japonicus, and compared the degree of genetic differentiation between them. High levels of polymorphism were observed over all loci. The mean alleles and expected heterozygosities over the seven stocks were 10.4–12.3 and 0.735–0.783 respectively. The results of the microsatellite survey provide no evidence to show that hatchery practice of the sea cucumber in China to date has significantly affected the genetic variability of the cultured stocks. Significant differentiation was found between most pairs of the hatchery stocks and wild populations (F_st range: 0.008–0.036), and no obvious difference was detected between the wild populations (F_st=0.008). The information on the genetic variation and differentiation obtained in this study can be applied for future genetic monitoring of A. japonicus aquaculture stocks and will be useful for future genetic improvement by selective breeding, and for designing suitable management guidelines for these genetic materials.     

饲料对吊笼养殖仿刺参生长及体成分的影响

在水温16.6~11.6℃、盐度29.3~27.6的海域环境条件下,将1 080 cfu初始体质量为(51.8±6.9)g的仿刺参(Apostichopus japonicus)放养于4组共36个网箱吊笼(规格为72 cm×39 cm×31 cm,分为6层),每个吊笼30 cfu(每层5 cfu),分别投喂海带(A组)、海带+鲜杂鱼糜(B组)、海带+扇贝边粉(C组)和仿刺参粉状配合饲料(D组),研究了不同饲料对仿刺参生长和体成分的影响。84 d的饲养结果表明,各处理组仿刺参成活率(SR)和脏壁比(VBWR)均差异不显著(P0.05),但增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)和蛋白质效率(PER)却不同。各组WGR、SGR分别介于172.3%~79.6%和(1.19~0.70)%·d-1之间,B、C两组WGR、SGR均差异不显著(P0.05),但显著高于A、D两组(P0.05),D组最小。B组FCR最低,D组最高,各处理组之间差异显著(P0.05)。A组PER最高,D组最低,各处理组之间差异显著(P0.05)。饲料影响仿刺参体成分,D组水分含量显著低于其它3组(P0.05),其它3组之间差异不显著(P0.05);各组粗蛋白含量变化在40.0%~48.1%之间,C组显著高于其它3组(P0.05),其它3组之间差异不显著(P0.05);粗脂肪含量变化较大,为1.3%~3.8%之间,B组最高,C组最低,各处理组之间差异显著(P0.05);A组灰分含量显著高于其它3组(P0.05),B组显著高于C、D两组(P0.05),而C、D两组之间无显著差异(P0.05)。综合仿刺参各生长指标,在试验饲料组中,饲喂海带与鲜杂鱼糜搭配饲料(B组)对网箱吊笼养殖仿刺参效果较好。