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.     

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.     

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.     

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 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.     

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.     

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.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

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.     

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 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.     

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.     

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.     

Dietary supplementation of red alga Pyropia spheroplasts on growth,feed utilization and body composition of sea cucumber,Apostichopus japonicus (Selenka)

A feeding experiment was conducted in a closed recirculating system to evaluate the effects of freeze‐dried spheroplasts prepared from Pyropia yezoensis (Ueda) on feed intake, growth and biochemical composition of sea cucumber, Apostichopus japonicus (Selenka). Pyropea spheroplasts (PS) were prepared through enzymatic treatment to break down the complex mixture of polysaccharides cell walls that might be easier for growth energy partitioning. Sea cucumbers were fed‐formulated diets with 10 (Diet 1), 30 (Diet 2) and 50 g/kg (Diet 3) inclusion level of PS. A diet without PS was used as a control (Diet 4). The experiment was conducted for 6 weeks maintaining water temperature 15 ± 1°C, photoperiod 18:06 hours (D:L). Feed was supplied ad‐libitum at 16.00 h once in a day, and the remaining feed and faeces were removed in the next day. Results showed that the highest growth was observed in the 50 g/kg PS diet compared to other treatments. Total weight gain, mean weight gain, net yield, protein efficiency ratio (PER) and protein gain (%) were significantly higher in the 50 g/kg PS diet (p < .05). A significantly higher percentage of energy was allocated for growth in the 50 g/kg PS diet. The highest specific growth rate and feed conversion efficiency (p < .05) were observed in the higher percentage of PS diet. Both the growth performance and biochemical analysis showed that superior growth was observed with increasing levels of PS in the diet. We infer that PS can be used as a new, cheaper feed ingredient in the formulated diet of A. japonicus.     

Effects of a probiotic mixture (Bacillus subtilis YB‐1 and Bacillus cereus YB‐2) on disease resistance and non‐specific immunity of sea cucumber,Apostichopus japonicus (Selenka)

The effect of a commercially available compound probiotics product containing Bacillus subtilis YB‐1 (50%) and Bacillus cereus YB‐2 (50%) fed to sea cucumbers, Apostichopus japonicus (Selenka) on challenge infections and non‐specific immune responses was assessed. Sea cucumbers (were randomly allocated into nine aquariums at a density of 30 sea cucumbers per tank and triplicate groups) were fed diets containing 0 (control), 10⁷ and 10¹⁰ cfu (g diet)^?1 of the probiotics mixture for 32 days. The growth factors and immunological parameters were measured. In addition, the effects on resistance against Vibrio alginolyticus infection were also evaluated. The results indicate that all the immunological parameters (phagocytic activity, superoxide anion production, lysozyme activity, catalase activity and phenoloxidase activity) measured and the growth rate of sea cucumbers fed 10¹⁰ cfu of the probiotics mixture were significantly (P < 0.05) improved than control groups at 16 and 32 days. After challenging, the cumulative mortality for the control was 100%, whereas the cumulative mortality for sea cucumbers fed 10¹⁰ cfu of the probiotics mixture was 47% (P < 0.05). Although the total autochthonous intestinal heterotrophic bacterial counts were not affected by dietary treatment (P > 0.05), Bacillus sp. levels were significantly elevated in sea cucumbers fed the probiotics mixture (P < 0.05). These results confirmed that administration of the probiotics mixture in the diet stimulated non‐specific immune responses and enhanced the growth performance of sea cucumbers, and was effective in controlling infections caused by V. alginolyticus.     

Effects of dietary n‐3 PUFA supplements on composition of n‐3 PUFA and expression of fatty acid elongase 5 (AJELOVL5) in sea cucumber,Apostichopus japonicus

This is the first comprehensive study on the effect of dietary polyunsaturated fatty acid (PUFA) levels on the expression of fatty acid elongase 5 (AJELOVL5), PUFA composition, and growth in juvenile sea cucumbers. The specific growth rate (SGR_w) was improved in n‐3 PUFA‐rich diets compared to low n‐3 PUFA diets. AJELOVL5 expression was apparently upregulated in juveniles fed lower PUFA diets relative to higher PUFA diets, with higher expression in the body wall and respiratory tree of juveniles fed diets without ɑ‐linolenic acid (ALA, 18:3n‐3) compared to juveniles fed higher ALA level diets; similar results were also detected in juveniles fed diets with lower eicosapentaenoic acid (EPA, 20:5n‐3), docosahexaenoic acid (DHA, 22:6n‐3), and none of ALA, EPA, or DHA respectively. The concentrations of ALA, EPA, and DHA in tissues were positively related to the content of dietary corresponding PUFA, with higher ALA content in juveniles fed diet ALA_12.71 than in the ALA_7.46 and ALA₀ groups. Similar results were also obtained in sea cucumber fed diets enriched with either EPA or DHA. Interestingly, considerable levels of EPA and DHA were found in the tissues of juveniles fed diets of CK₀ and DHA₀, with no specific input of EPA or DHA, showing that the sea cucumber was capable of biosynthesizing EPA and DHA from their corresponding precursors as ALA and linoleic acid (LA, 18:2n‐6).     

Effect of dietary chitosan oligosaccharide complex with Ce (IV) on growth,immunity and disease resistance against Vibrio splendidus of sea cucumber,Apostichopus japonicas

An 8‐week feeding trial was conducted to investigate the effects of dietary chitosan oligosaccharide complex with cerium (Ce IV) (COS‐Ce) on growth performance, nonspecific immunity and disease resistance of sea cucumber, Apostichopus japonicas. Five isonitrogenous (18.6%) and isolipidic (1.1%) practical diets were formulated with graded level of COS‐Ce (0, 150, 300, 600 and 1200 mg kg^?1 dry feed), which were named as COS‐Ce/0, COS‐Ce/150, COS‐Ce/300, COS‐Ce/600, COS‐Ce/1200 respectively. Each diet was allocated to four replicates of sea cucumbers (Initial weight: 6.72 ± 0.02 g). Sea cucumbers were fed to apparent satiation once daily (19:00 hours) for 56 days. During the experiment, water temperature was kept at 16 ± 0.5°C, pH 7.8–8.2, dissolved oxygen beyond 5 mg L^?1, ammonia nitrogen below 0.5 mg L^?1 and salinity from 30‰ to 31‰. Results showed that the specific growth rate of sea cucumbers was significantly higher in COS‐Ce/600 than that in other four treatments. Activities of phagocytosis, respiratory burst, acid phosphatase and alkaline phosphatase in COS‐Ce/600 were significantly higher than that in COS‐Ce/0 (P < 0.05) respectively. On the contrary, cumulative mortality was the lowest in COS‐Ce/600 following 14 days exposure to Vibrio splendidus (P < 0.05). In conclusion, these results confirmed that dietary COS‐Ce had beneficial effects on growth performance, nonspecific immunity and disease resistance of sea cucumber.     

Immune responses of the sea cucumber Apostichopus japonicus to stress in two different transport systems

We report the effect of transport stress on the immune responses of the sea cucumber Apostichopus japonicus. The total coelomocytes counts, phagocytosis activity, reactive oxygen species production, acid phosphatase activity, phenoloxidase activity and catecholamine levels were measured in A. japonicus following both damp and water transportation methods. With both transport methods, stress resulted in significant elevation of catecholamine levels. Total coelomocytes counts increased following transportation, whereas phagocytic activity and reactive oxygen species production decreased. Damp transport significantly depressed acid phosphatase and phenoloxidase activity in A. japonicus. However, there were no significant differences in acid phosphatase and phenoloxidase activity after water transportation. These results show that transport stress induces modifications in the immune system of A. japonicus and that damp transport elicited more severe stress than water transport. This study increased understanding of the mechanism of transport stress inducing neuroendocrine responses and immune modification in A. japonicus. The information obtained should be helpful to optimize transport process in sea cucumber culture.     

Use of Nannochloropsis sp. isolated from the East China Sea in larval rearing of sea cucumber (Apostichopus japonicus)

The supply of microalgae to hatcheries is a limiting factor for the mass larval production of sea cucumber (Apostichopus japonicus) in Fujian Province, China. In this study, Nannochloropsis sp. isolated from the East China Sea was tested as food for A. japonicus larvae. The first trial compared the effect of mono‐, bi‐ and trialgal diets comprising three microalgae (Chaetoceros muelleri, Dunaliella tertiolecta and Nannochloropsis sp.) on A. japonicus larval growth, survival, settlement and juvenile growth. The results showed that there were no significant differences in survival and settlement between larvae fed with Nannochloropsis sp. and other diets. All diet treatments yielded similar juvenile sea cucumber output. In the second trial, A. japonicus larvae were fed equally four times daily at three different rations (5000, 20 000 and 40 000 cells mL^?1 day^?1). Larvae fed 20 000 cells mL^?1 day^?1 were significantly larger than larvae in other groups and experienced the highest survival rate. In the third trial, A. japonicus larvae were fed 20 000 cells mL^?1 day^?1 in three different frequency (2, 3 and 4 meals day^?1). The greatest body length was observed in larvae that received 3 meals day^?1. Survival and settlement of larvae fed 2 meals day^?1 were significantly lower than other two groups. These results suggest that Nannochloropsis sp. can be used as a diet for the large‐scale production of A. japonicus seed, and larvae fed three times daily at a ration of 20 000 cell mL^?1 day^?1 are recommended for hatchery production of A. japonicus.     

饲料添加半乳甘露寡糖对刺参幼参生长、体壁营养组成及免疫力的影响

以初始体质量为(1.79±0.06) g的刺参(Apostichopus japonicus)为研究对象, 进行为期8周养殖实验, 研究饲料中添加半乳甘露寡糖对刺参生长、消化、体壁营养组成及免疫力的影响。以基础饲料(E0)为对照组, 在基础饲料中分别添加0.2% (E1)、0.4% (E2)、0.8% (E3)、1.2% (E4)和1.6% (E5)的半乳甘露寡糖, 共配制5种实验饲料。结果表明: (1) 饲料中添加不同浓度的半乳甘露寡糖对幼参增重率及特定生长率具有显著影响, 各处理组均显著高于对照组(P<0.05), 而对脏壁比及肠壁比无显著影响(P>0.05), 1.2%(E4组)添加水平下, 刺参增重率及特定生长率最高。(2) 饲料中添加不同浓度的半乳甘露寡糖对幼参肠道蛋白酶、淀粉酶、纤维素酶活力均无显著影响(P>0.05)。(3) 饲料中添加不同浓度的半乳甘露寡糖对幼参体壁酸性黏多糖及糖醛酸含量无显著影响(P>0.05)。添加半乳甘露寡糖比例0.2%(E1)、0.4%(E2)、0.8%(E3)、1.2%(E4)的处理组, 刺参体壁氨基酸总量及谷氨酸、甘氨酸、精氨酸含量显著高于对照组(P<0.05), 而1.6%(E5)组体壁氨基酸总量及Glu、Gly、Arg含量与对照组无显著差异(P>0.05)。(4) 饲料中添加半乳甘露寡糖对体壁及体腔液超氧化物歧化酶、碱性磷酸酶及溶菌酶活性具有显著影响(P<0.05), 同时对体腔细胞溶菌酶活性影响显著(P<0.05)。实验可以得到以下结论: (1) 饲料中添加半乳甘露寡糖不仅可以提高刺参免疫力, 并且可以提高特定生长率, 并改善体壁氨基酸组成; (2) 刺参饲料中半乳甘露寡糖的合适添加比例为1.2%。