Effect of diet on growth and body biochemical composition of juvenile four‐sided sea cucumber Isostichopus badionotus (Selenka, 1867)

The farming of sea cucumber is a growing industry that demands feeds that are tailored to the organism's nutritional needs. Therefore, in this study, the effect of diet composition, with and without additives, on the growth, survival, specific growth rate (SGR) and body biochemical composition of the four‐sided sea cucumber Isostichopus badionotus was evaluated over a period of 3 months. Four diets were used: D1 (Macrocystis sp. and Sargassum sp.), D2 (Macrocystis sp. and Spirulina sp.), D3 (Sargassum sp. and Spirulina sp.) and D4 (Macrocystis sp., Sargassum sp. and Spirulina sp.), and the same diets enriched with 0.22% of Algamac 2000^® (D1H, D2H, D3H and D4H) were also tested. The organisms fed the diets containing Spirulina sp. demonstrated the highest growth rates and significantly greater SGR (% per day) (p < .05), an effect that was enhanced by the food additive (D2 0.67%; D2H 0.78%; D3H 0.76%). Survival and body composition were not affected by diet type. Based on these results, the use of Spirulina sp. combined with supplements rich in PUFA content is recommended in the formulation of diets to promote the growth of I. badionotus juvenile.     

Spawning and Larval Development of the Four‐Sided Sea Cucumber,Isostichopus badionotus (Selenka 1867), under Controlled Conditions

The objective of this study was to generate information for the development of Isostichopus badionotus culture protocols by determining optimal spawning methods, egg production, length and duration of larval stages, and larval habitat settlement preferences. Studies performed during the spawning seasons of 2010 and 2011 showed that this organism spawns between July and November without mechanical or chemical induction. The larval development has five well‐defined stages: early, mid and late auricularia, doliolaria, and pentactula. Juveniles (654.3 µm length) were obtained on average 25 days after fertilization (DAF), when larvae were incubated at 25 ± 1 C. No preference was observed for any of the materials used for settlement of the larvae. The easy adaptation of I. badionotus to the culture conditions and the competence of the larvae to complete metamorphosis while feeding solely on microalgae concentrates make this species an excellent candidate for farming. However, it is necessary to continue designing techniques for larval culture and to establish appropriate captivity conditions that allow multiannual reproduction.     

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.     

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.     

Dietary supplements of guanosine improve the growth,non‐specific immunity of sea cucumber,Apostichopus japonicus Selenka,and its resistance against Vibrio splendidus

The growth response, non‐specific immune activities and disease resistance were measured in sea cucumber, Apostichopus japonicus Selenka (initial average weight 6.80 ± 0.10 g), when fed diets supplemented with graded levels of guanosine from the guanosine‐5’‐monophosphate disodium (GMP) at 0 (control), 0.3, 0.6 and 1.2 g/kg for 8 weeks. The results showed that GMP supplemented at 0.6 and 1.2 g/kg significantly enhanced the growth of sea cucumber. Sea cucumber fed a diet with 0.6 g/kg of GMP had significantly higher intracellular superoxide anion production, nitric oxide synthase activity, lysozyme activity and the total superoxide dismutase (T‐SOD) activity than those in control group (p < .05). Increased lysozyme activity and T‐SOD activity were also found in sea cucumber fed GMP at 1.2 g/kg. Moreover, there was significantly lower cumulative mortality after the disease challenge in sea cucumber fed the diets with 0.6 and 1.2 g/kg GMP than that in control and 0.3 g/kg GMP groups (p < .05), and no significant difference was observed between 0.6 and 1.2 g/kg GMP groups. These results suggested that feeding GMP at a dose of 0.6 g/kg could enhance growth, non‐specific immunity of sea cucumber as well as its resistance against Vibrio splendidus.     

Effects of rearing temperature on growth,metabolism and thermal tolerance of juvenile sea cucumber,Apostichopus japonicus Selenka: critical thermal maximum (CTmax) and hsps gene expression

Effects of different rearing temperatures (16, 21 and 26°C) on growth, metabolic performance and thermal tolerance of juvenile sea cucumber Apostichopus japonicus (initial body weight 7.72 ± 0.96 g, mean ±SD) were investigated in this study. During the 40‐day experiment, growth, metabolic performance, food intake and energy budget at different reared temperatures were determined. Sea cucumbers rearing at 16°C obtained better growth (final body weight 11.96 ± 0.35 g) than those reared at 21 (10.33 ± 0.41 g) and 26°C (8.31 ± 0.19 g) (P < 0.05), and more energy was allocated for growth at 16°C (162.73 ±11.85 J g^?1 d^?1) than those at 21(79.61 ± 6.76 J g^?1 d^?1) and 26°C (27.07 ± 4.30 J g^?1 d^?1) (P < 0.05). Critical thermal maxima (CTmax) values of juvenile sea cucumbers reared at 16, 21 and 26°C were 33.1, 34.1 and 36.6°C, respectively, and the upregulation of hsps in sea cucumbers reared at 26°C was higher than those acclimated at lower temperatures (16 and 21°C), indicating that temperature acclimation could change the thermal tolerance of the sea cucumber, and CTmax and hsps were sensitive indicators of the sea cucumber's thermal tolerance.     

Effects of diatom concentration in prepared feeds on growth and energy budget of the sea cucumber Apostichopus japonicus (Selenka)

Eight test diets were used in the experiment: seven fresh diatom (Cylindrotheca fusiformis) diets in which diatom occupied 100%, 83.3%, 50%, 33.3%, 20%, 14.3% and 11.1% in dry matter of the feeds (marked as D100, D83, D50, D33, D20, D14 and D11), and a formulated diet (70% Sargassum thunbergii powder, 20% sea mud and 10% fish meal, based on dry weight, marked as ST). The results showed that the specific growth rates (SGRs) of the animals fed diets D33, D20, D14 and D11 were not significantly different from the SGR of those fed diet ST. However, the energy allocated to growth for the animals fed with diet D14 was twice that of diet ST. For sea cucumbers fed diet D14 the largest expenditure part was allocated to respiration (55.4%), but for those fed diet ST, the largest part was allocated to faeces (62.7%). The organic content of diet D14 was also much lower than that of diet ST. Frozen diatom diets containing 14% (dry matter weight) fresh diatom could be an environmentally friendly feed as an alternative option for macroalgae powder.     

Dietary supplement of fructooligosaccharides and Bacillus subtilis enhances the growth rate and disease resistance of the sea cucumber Apostichopus japonicus (Selenka)

The effects of two dietary supplements, entrapped fructooligosaccharide (FOS) and microencapsulated Bacillus subtilis, on the growth and disease resistance of the sea cucumber Apostichopus japonicus were investigated. Sea cucumbers (a total of 810 animals, with initial weight of 3.72 ± 0.16 g each) were fed with basal diet only or basal diet supplemented with three different doses of B. subtilis (0.2, 0.4 and 0.8%), three different doses of FOS (0.4, 0.8 and 1.6%), 0.2% B. subtilis plus 0.4% or 0.8% FOS. Growth and lysozyme activity were measured after 50 days feeding. The results showed that dietary 0.2% and 0.4% B. subtilis alone or 0.2% B. subtilis plus 0.8% FOS significantly increased the specific growth rate (SGR) of sea cucumbers compared with control (P < 0.05). The addition of B. subtilis with or without FOS had no significant effect on lysozyme activity. The animals were also challenged with Vibrio splendidus to assess the level of disease resistance provided by the two dietary supplements. Sea cucumbers fed with 0.8% B. subtilis or 0.2% B. subtilis plus 0.8% FOS exhibited the highest survival (P > 0.05). Under these experimental conditions, dietary B. subtilis and FOS had no synergistic effect on enhancing SGR and disease resistance of sea cucumber.     

Influence of temperature on growth and survival of juvenile winter flounder,Pseudopleuronectes americanus (Walbaum) reared under continuous light

Winter flounder, Pseudopleuronectes americanus, has emerged as a promising candidate flatfish for cold‐water aquaculture and restocking. Here, juveniles were reared for 8 weeks at three temperatures: 10, 15, and 20°C under 24‐hr light. All fish were imaged at stocking and at 2‐week intervals, where growth was measured as changes in standard length (S_L) and body area (B_A). By week 2, fish reared at 15 and 20°C were larger than those grown at 10°C. At weeks 4 to 6, fish at 15°C were larger than fish at 20°C. Linear regressions were used to model growth dynamics over time at each temperature. Highly significant linear growth trajectories were detected over time for S_L and B_A. S_L and B_A regressions also showed a significant difference among the slopes across temperatures, where comparing slopes showed the best temperature to rear the flounder was 15°C. Weights of fish held at 15°C and 20°C were greater than at 10°C at the termination of the experiment. Within each temperature, the growth rate of malpigmented fish was not different from that of the normally pigmented fish. Overall, growth of winter flounder was comparable to that of other commercially produced flatfish species, providing strong evidence for this flatfish species as a potential species for aquaculture.     

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C_maxe; J day^?1) and the specific growth rate in terms of energy (SGRe; % day^?1) in animals of three body sizes (mean±SE) – large (134.0±3.5 g), medium (73.6±2.2 g) and small (36.5±1.2 g) – were determined at water temperatures of 10, 15, 20, 25 and 30°C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20°C. This trend, however, was not apparent at 25 and 30°C, which could be influenced by aestivation. High water temperatures (above 20°C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15°C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3–139.3 g) was 24.5?25.5°C, while that for small animals (28.9–40.7 g) was between 25.5 and 30.5°C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.     

Effects of dietary heat‐killed Lactobacillus plantarum L‐137 (HK L‐137) on the growth performance,digestive enzymes and selected non‐specific immune responses in sea cucumber,Apostichopus japonicus Selenka

A feeding trial was conducted to evaluate the efficacy of dietary heat‐killed Lactobacillus plantarum L‐137 (HK L‐137) on growth performance, digestive, non‐specific immune and phagocytosis of sea cucumber, Apostichopus japonicus. Sea cucumbers (initial body weight 1.35 ± 0.04 g) were fed diets supplemented with five levels of HK L‐137 (0, 0.005, 0.025, 0.05 and 0.25 g HK L‐137 kg^?1 diets) for 60 days. Results indicated sea cucumbers fed with diets containing 0.05 and 0.25 g HK L‐137 kg^?1 diets showed significantly (P < 0.05) higher body weight gain and specific growth rate than other groups. Sea cucumbers fed with diets containing 0.05 and 0.25 g HK L‐137 kg^?1 diets showed significantly (P < 0.05) higher protease activity than control group. Higher amylase, lysozyme and phagocytic activities were found in 0.25 g HK L‐137 kg^?1 diet group. Higher superoxide dismutase enzyme and alkaline phosphatase activity was found in 0.05 g HK L‐137 kg^?1. While no significant differences (P > 0.05) were found in acid phosphatase activity. These results suggested that dietary supplementation of 0.05 g HK L‐137 kg^?1 diets would have benefit on growth, digestive enzymes and several non‐specific immune parameters of sea cucumber.     

Effects of differently processed diets on growth,immunity and water quality of the sea cucumber,Apostichopus japonicus (Selenka, 1867)

The type of formulated diet used in aquaculture systems affects the feed efficiency and water quality. In this study, the water quality, growth performance and immunity of the sea cucumber, Apostichopus japonicus (Selenka, 1867), fed diets produced using four different processing methods (extruded pellet diet, cold‐bonded pellet diet, flake diet and mash diet) were quantified for 60 days to identify the optimal feed type for culturing of this species. Sea cucumbers grew faster, the feed conversion ratio was much lower, and immunity indicators were higher when fed the extruded pellet diet compared to the other diets. Sea cucumbers fed the extruded pellet diet had the highest specific growth rate (0.96), whereas A. japonicus fed the cold‐bonded pellet diet had the lowest specific growth rate (0.51). The lysozyme activity of sea cucumbers fed the extruded pellet diet was higher (57.52) than that of A. japonicus fed the cold‐bonded pellet diet (39.22), flake diet (37.32) and mash diet (32.68). When animals were fed the mash diet, the ammonia–nitrogen, nitrite–nitrogen and phosphate productions were higher than in animals fed the other diets. These results indicate that the extruded pellet diet is the optimum feed type for use in the culture of A. japonicus.     

Effect of dietary carbohydrate levels on growth performance,non‐specific immune enzymes and acute response to low salinity and high temperature of juvenile sea cucumber Apostichopus japonicus

This study was performed to determine the optimum dietary carbohydrate (CHO) levels of sea cucumber, based on the parameters of growth, digestive enzymes, digestibility, non‐specific immune enzymes and acute low‐salinity (20 g/L) stress and high‐temperature (30°C) stress tolerance. Diets with eight different CHO (dextrin) levels (32.9, 107.6, 192.5, 257.2, 316.8, 428.0, 482.4 and 572.8 g/kg) were fed to sea cucumber juveniles (0.49 ± 0.01 g) for 60 days. Significant higher amylase activity was observed in sea cucumbers fed diet with CHO ranging between 32.9 and 192.5 g/kg than that of other treatments (p < .05). The sea cucumbers fed with 192.5 g/kg CHO showed significantly higher acid phosphatase activity than the treatments of 482.4 and 572.8 g/kg CHO (p < .05), and significantly higher alkaline phosphatase activity than other treatments (p < .05, except 257.2 g/kg). The treatments of 428.0–572.8 g/kg were found significantly lower values than other treatments in apparent digestibility coefficients for dry matter and crude protein (p < .05). The sea cucumbers fed with 192.5, 257.2 and 316.8 g/kg CHO showed better tolerance to high‐temperature (30°C) and low‐salinity (20 g/L) stress than other treatments. In brief, the optimal dietary CHO level for the growth of juvenile sea cucumber is 177.96 g/kg. However, excessive CHO will inhibit amylase enzyme activity and decrease digestibility, resulting in low growth of sea cucumber.     

Feeding and growth in captivity of the octopus Enteroctopus megalocyathus Gould, 1852

The effect of diet on the growth and survival in captivity of the octopus Enteroctopus megalocyathus Gould, 1852 was researched to establish culture potential. This is one of two commercially important octopus species that inhabit the Chilean coast. A positive energy budget, averaging 678 J day^?1 g of dry weight (dw), was recorded in specimens maintained in tanks and fed ad libitum on a diet of three species of crustacean decapods. On the other hand, the energy budget of octopus specimens fed on the mussel Mytilus chilensis (Hupé) was negative, averaging ?250.90 J day^?1 g dw^?1. Ingestion rates varied between diets (t=46.45; d.f.=18; P<0.001), but no variations were recorded in energy loss resulting from metabolism (t=0.67; d.f.=16; P>0.05) or ammonia excretion (t=1.39; d.f.=16; P>0.05). An average instantaneous growth rate (IGR) of between 1.96% and 0.49% was recorded in specimens fed on crustaceans, depending on the size of the octopus and the period of time. The IGR in specimens fed on mussels was ?0.32±0.05. The IGR of specimens fed on crustaceans and maintained in suspended systems was 1.36±0.9%. The mean mortality with crustacean and mussel diets was 11.07±5.5% and 74±1% respectively. The results indicate that E. megalocyathus can survive and grow in captivity, both in tanks and in suspended systems, when fed on a diet of crustaceans.     

Effect of stocking density on growth performance of juvenile sea cucumber Holothuria tubulosa (Gmelin, 1788)

Stock density is one among the most important factors in aquaculture that directly influences the growth of organisms; however, there is limited information about the effects of stocking density on growth performance of sea cucumber Holothuria tubulosa (Gmelin, 1788) that has a commercial potential and represents a new species for aquaculture. In this study, we investigated the effect of stocking densities on the growth performance of juvenile H. tubulosa (40.3 ± 3.34 g) in laboratory conditions. Stocking densities were selected as 6, 15 and 30 ind m^?2 with total biomass 253.3 ± 0.18, 601.2 ± 0.11 and 1201.4 ± 0.15 g m^?2 respectively. We monitored the growth by wet weights and calculated the growth performance through weight gain, growth rate, specific growth rate, relative weight gain, coefficients of variation and survival rate during 8 weeks of research period. We found that individual mean weight gain in 6 ind m^?2 group was approximately 29.53 g while it was 3.03 g and ?4.36 g for 15 ind and 30 ind m^?2 groups respectively. Results have shown that the specific growth rate tends to decrease as the number of individuals in unit area increases. The final mean weight, growth rate, specific growth rate, relative weight gain and coefficients of variation were significantly different among treatment groups. In conclusion, a density of 6 ind m^?2 is recommended for stocking juvenile H. tubulosa under rearing conditions. Stocking density of 15 ind m^?2 is not favourable for long term as the growth rate is negligible where 30 ind m^?2 should be avoided in tank‐based rearing units.     

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.     

Growth,immunity and ammonia excretion of albino and normal Apostichopus japonicus (Selenka) feeding with various experimental diets

An experiment was conducted to evaluate the effects of six experimental diets on growth performance, ammonia excretion and immunity of albino and normal Apostichopus japonicus. A factorial design was used, the factors being type of diets (six levels) and colour of A. japonicus (two levels). A total of 30 randomly selected albino A. japonicus were housed in each (60 × 50 × 30 cm³) of 18 blue plastic aquaria to form six groups in triplicate, and the same set‐up was used for the normal A. japonicus. Each group of animals was fed with one of the six experimental diets. Apparent dry matter digestibility (ADMD) and apparent crude protein digestibility (ACPD) were analysed using acid‐insoluble ash (AIA) content method. At the end of the experiment, all A. japonicus were harvested and weighed to calculate growth parameters. After weighing, six individuals from each aquarium were randomly sampled for immune indices. Results indicated that all growth parameters of A. japonicus increased with decreasing nutrient content in their diets (p < .01), whereas an opposite result was observed in case of the ammonia‐nitrogen production by A. japonicus. Normal A. japonicus grew better (p < .01) and produced lower (p < .01) quantity of ammonia nitrogen compared to the albino A. japonicus. Immunity particularly superoxide dismutase and lysozyme activities was higher (p < .05) in normal compared to albino A. japonicus. Considering all measured variables, D1 (diet containing crude protein, crude lipid, carbohydrate and crude ash 51.8, 8.7, 231.3, 708.2 g/kg, respectively) was the best diet among all experimental diets. More research is still needed to optimize nutrients in the diet of A. japonicus, as this study does not provide information about critical threshold level of nutrients in diets. Until then, diet D1 can be recommended for A. japonicus aquaculture.