Nitrogen and phosphorus budget of a Haliotis discus hannai and Apostichopus japonicus polyculture system

The system nitrogen (N), phosphorous (P) budget, N/P utilization rate and the physiological response mechanism of the abalone Haliotis discus hannai (body weight: 12.87 ± 0.82 g) and the sea cucumber Apostichopus japonicus (body weight: 10.85 ± 1.16 g) to different co‐culture environment conditions were examined. Animals were kept in a multilayer, cubic recirculating aquaculture system at different polyculture densities (abalones at 400 ind/m²in monoculture [Group C] and abalones at 400 ind/m²with sea cucumbers at 10 ind/m² [AS1] or 20 ind/m² [AS2]). Each treatment was replicated four times, and the experimental cycle was 90 days. No significant difference in survival rate of abalones was detected when the stocking density of sea cucumbers increased from 10 to 20 ind/m², but the concentrations of total ammonia nitrogen and nitrite nitrogen in the water were significantly higher in AS2 than in AS1. Survival rate, specific growth rate (SGR) of body weight of sea cucumbers and SGR of body weight of abalones were significantly lower in AS2 than in AS1. No significant difference in protease (PES), lipase, amylase and cellulase activities of abalones was identified between Group C and AS1, but the PES and amylase activities of abalones and sea cucumbers in AS1 were significantly higher than those in AS2. In AS1, the N/P output from harvesting of abalones and sea cucumbers and the N/P utilization rates were significantly higher than those in AS2. Although the N/P output from faeces was significantly lower in AS2 than in Group C and AS1, the N/P output from the water layer was significantly higher than that in AS1. The expression levels of Cu/Zn superoxide dismutase (Cu/Zn‐SOD) and heat shock protein 70 (HSP70) of both abalones and sea cucumbers in AS2 were significantly higher than those in AS1. No significant difference in expression of catalase (CAT) and HSP90of abalones was identified among these groups, but the expression levels of CAT and HSP90of sea cucumbers in AS2 were significantly higher than those in AS1. These results indicate that stocking sea cucumbers at 10 ind/m² in the polyculture system will relieve the organic load on the system and improve the N/P utilization rate. It will also increase aquaculture production and improve the ecological and economic benefits of the system.     

Effects of flow velocity on growth,metabolism and nitrogen and phosphorus budget of Haliotis discus hannai Ino in a recirculating aquaculture system

To study the effects of different flow velocities on growth, metabolism and nitrogen (N) and phosphorus (P) budget of Haliotis discus hannai Ino in a recirculating aquaculture system, abalones (shell length: 45.71 ± 3.09 mm, body weight: 8.62 ± 0.75 g) were cultured at three different flow velocities with four replicates each: high (400 L/hr), medium (300 L/hr) and low (200 L/hr). At the end of the 70 days experiment, the survival rate, food intake and P utilization rate of abalones in the 200 L/hr group were significantly lower than those in any other group (p < .05), whereas the total ammonia nitrogen and nitrite nitrogen concentrations in the water were significantly higher than those in any other group (p < .05). The body weight specific growth rate, food conversion efficiency, abalone‐harvested N and P outputs and N utilization rate were ranked as follows: 300 > 400 > 200 L/hr (p < .05). In the 300 L/hr group, the activities of hexokinase, pyruvate kinase and lactate dehydrogenase were significantly lower than those in the 400 L/hr group, whereas the activity of succinate dehydrogenase was significantly higher than that in the other two groups (p < .05). Aerobic metabolism was the main source of energy acquisition for abalones in the 300 L/hr group. In the 400 L/hr group, the expression levels of manganese superoxide dismutase and thioredoxin peroxidase significantly increased from Day 35 to Day 70, but they still were significantly lower than the values in the 200 L/hr group (p < .05). In the 200 L/hr group, the expression levels of catalase and heat shock protein 70 were significantly higher than those in any other group, and they significantly increased from Day 35 to Day 70 (p < .05). These results indicate that maintaining flow velocity at 300 L/hr will minimize oxidative damage due to deterioration of water quality, reduce the energy consumption required for organisms to resist the impact of water flow, improve the N and P utilization rates of the recirculating aquaculture system, and reduce the cost of culturing abalones.     

Hybridization between Haliotis rufescens and Haliotis discus hannai: evaluation of fertilization,larval development,growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ × J♂ showed a fertilization rate of 55.3 ± 3.5% using 20‐min‐old eggs and sperm concentrations of 14 × 10⁶ cells mL^?1, while the reciprocal cross (J♀ × R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.     

Parentage assignment in hybrid abalones (Haliotis rufescens × Haliotis discus hannai) based on microsatellite DNA markers

Parentage analysis in aquaculture determines genealogical relationships between broodstock and progeny when the parents are unknown. Thus, parentage analysis is a useful tool to establish pedigree reports in molecular‐assisted selection programs. Here, we evaluated 10 heterologous microsatellite markers for parentage assignment in abalone hybrids produced from 43 abalone broodstocks of red abalone (Haliotis rufescens) and Japanese abalone (H. discus hannai). The allele frequencies, exclusion probabilities and broodstock contributions were calculated using CERVUS, PAPA and GERUD software. The polymorphic information content (PIC) values showed that most of the microsatellite loci were highly informative (>0.7) and more than 90% of parentage assignment was possible with a minimum of 5–6 microsatellite markers. Parentage assignment for hybrid and pure‐red progeny showed a better performance than pure‐Japanese progeny. This result could be due to the high level of allele loss in the parental genotypes. In addition, results indicated that only two sires contributed over 80% and 90% of red and hybrid progenies, respectively. This study gives a new molecular tool to support marker‐assisted selection in abalone hybrids produced in Chile.     

Protective effects of dietary selenium on abalone Haliotis discus hannai against the toxicity of waterborne cadmium

This study was conducted to investigate protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of cadmium (Cd). A 60‐day feeding trial was conducted with abalone (initial weight: 3.17 ± 0.01 g), which were exposed to 0.34 mg/L of waterborne Cd. During a feeding trial, abalone were fed graded levels of Se at 0.10 (controls), 1.31, and 4.20 mg/kg diet respectively. Results showed that there was no significant difference in specific growth rate and survival rate of abalone among the three treatments. Compared with the controls, dietary Se significantly decreased Cd concentrations in serum, muscle, mantle, gill, mantle, and hepatopancreas of abalone. Besides, compared with the controls, dietary Se significantly increased metallothionein concentration in the hepatopancreas of abalone. Additionally, compared with the controls, dietary Se significantly decreased concentrations of malondiadehyde and protein carbonyl in hepatopancreas of abalone. Meanwhile, compared with the controls, dietary Se significantly increased activities of glutathione peroxidase, thioredoxin reductase, and thioredoxin peroxidase, and concentration of glutathione in the hepatopancreas of abalone. Based on the data above, in abalone, dietary Se showed protective effects against waterborne Cd.     

Dose‐dependent protective effects of dietary selenium on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper

This study was designed to assess the protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper (Cu). A 60‐day feeding trial was conducted in a static water system for abalone (initial weight: 3.17 ± 0.01 g) exposed to 0.02 mg L^?1 of waterborne Cu. The animals were fed one of the three experimental diets with 0.10, 1.31 and 4.20 mg kg^?1 of Se from Na₂SeO₃·5H₂O respectively. Results showed that the abalone fed 1.31 mg kg^?1 of dietary Se had the lowest Cu concentration in shell, muscle, mantle, gill, hepatopancreas and serum. Meanwhile, the significant lowest contents of malondiadehyde and protein carbonyl in hepatopancreas were also found in the treatment with 1.31 mg kg^?1 of dietary Se (P < 0.05). In addition, this treatment had significant higher glutathione content and thioredoxin reductase activity in hepatopancreas (P < 0.05). However, the activity of Se‐dependent glutathione peroxidase (Se‐GPx) was significantly decreased in the treatment with 4.20 mg kg^?1 of dietary Se (P < 0.05). In this treatment, the protein carbonyl content in hepatopancreas was significantly higher than that in the group with 1.31 mg kg^?1 of dietary Se (P < 0.05). In conclusion, in terms of anti‐oxidation and Cu accumulation, the protective effects of dietary Se on abalone against waterborne Cu were dose‐dependent. The 1.31 mg kg^?1 of dietary Se had this effect, but not 4.20 mg kg^?1 of dietary Se. Moreover, the latter increased the oxidative stress in abalone exposed to the waterborne Cu.     

Influences of dietary antimicrobial peptide APSH‐07 on the growth performance,immune response and vibriosis resistance of abalone Haliotis discus hannai Ino

A 115‐day feeding trial and subsequently a 10‐day challenge test with Vibrio parahaemolyticus were conducted to investigate the effects of dietary antimicrobial peptide APSH‐07 on growth performance, immune response, antioxidative status and vibriosis resistance of abalone Haliotis discus hannai (initial body weight: 2.06 ± 0.01 g; initial shell length: 25.42 ± 0.18 mm). Four artificial diets were designed with 0 (artificial diet control), 7.5, 15.0 and 22.5 mg/kg of APSH‐07, respectively. The brown alga Laminaria japonica was used as the live food control. Results showed that the specific growth rates of abalone in the groups with 7.5 and 15.0 mg/kg dietary APSH‐07 were significantly higher than those in the controls (p < .05). The total haemocyte counts and respiratory burst activity in haemolymph of abalone in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the groups with 0 and 22.5 mg/kg of dietary APSH‐07 (p < .05). The gene expression levels of Mn‐superoxide dismutase and thioredoxin peroxidase 2, and the activities of glutathione peroxidase in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the other groups (p < .05). Cumulative mortality of abalone after the challenge test was significantly decreased in the group with 7.5 mg/kg of dietary APSH‐07 supplementation. Supplementation of 22.5 mg/kg dietary APSH‐07 significantly increased the cumulative mortality. In conclusion, 7.5 mg/kg of dietary APSH‐07 supplementation had the better growth performance, higher antioxidation, immune and disease resistance capacity of abalone. Excessive supplementation of dietary antimicrobial peptide APSH‐07 (22.5 mg/kg) had significantly negative effects. Further studies are needed to determine the optimal level of dietary APSH‐07 supplementation for abalone.     

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.     

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.     

基于线性食物网模型估算荣成俚岛人工鱼礁区刺参和皱纹盘鲍的生态容纳量

俚岛人工鱼礁区是中国北方近海典型的海珍品增殖型人工鱼礁生态系统,刺参(Apostichopus japonicus)和皱纹盘鲍(Haliotis discus hannai)是该生态系的主要增殖放流种类。基于2009年在荣成俚岛人工鱼礁区进行的周年生物资源调查数据,利用EWE软件构建俚岛人工鱼礁区生态系统生态通道模型(Ecopath),对该模型进行了不确定性及输入参数敏感度检验,系统分析了俚岛人工鱼礁区生态系统特征和能量流动规律,估算了俚岛人工鱼礁区生态系统内刺参和皱纹盘鲍的生态容纳量。模型由19个功能组构成,基本涵盖了俚岛人工鱼礁区生态系统能量流动的主要过程。分析结果表明,模型可信度P指数为0.51,小型底层鱼类生产量与生物量的比值(P/B)下降50%会使小型底层鱼类生物量(B)估算值变化1.36,影响最大;系统总流量为10 786.680 t/(km2.a),其中27%流向碎屑,17%以捕捞和沉积的形式流出系统;系统的总初级生产力为4 131.966 t/(km2.a);系统的总能量转换效率为10.5%,流量中来自碎屑的比例为39%,其中的61%直接来源于初级生产者,能流通道以牧食食物链为主导;刺参和皱纹盘鲍的生态容纳量为309.4 t/km2和198.86 t/km2,目前礁区刺参和皱纹盘鲍的生物量分别占估算生态容纳量的31.72%和26.15%,仍具一定的增殖空间。     

Effects of dietary protein levels on the activity of the digestive enzyme of albino and normal Apostichopus japonicus (Selenka)

We evaluated the effects of different proportions of dietary protein (5%, 10%, 15%, 20%, 25% and 30% protein) on the activity of digestive enzymes of normal and albino Apostichopus japonicus. The experimental diets were fed for 60 days, the optimal conditions for digestive enzyme activity in sea cucumbers were studied. The optimal temperature for protease was 29.3°C and the optimal pH was 1.8. The optimal temperature for amylase was 34.3°C and the optimal pH was 6.7. The optimal temperature for cellulase was 56°C and the optimal pH was 5.9. The activity of intestinal protease increased at first and then decreased as the proportion of dietary protein increased, reaching the maximum when the proportion of protein was 19.7%. The activity of protease in the intestine of normal sea cucumber was significantly lower than that of albino sea cucumber, and the activity of amylase was significantly higher than that of albino sea cucumber. This study is expected to provide a basis for further explaining the ecological difference of albino and normal A. japonicus.     

A study of organic carbon,nitrogen and phosphorus budget in jellyfish–shellfish–fish–prawn polyculture ponds

A budget describing the flow of organic carbon (OC), nitrogen (N) and phosphorus (P) through two polyculture ponds (jellyfish–shellfish–fish–prawn) was constructed. The total input of OC was 3107 kg ha^?1 in pond 1 and 3358 kg ha^?1 in pond 2, while total output was 1759 kg ha^?1 in pond 1 and 1325 kg ha^?1 in pond 2. In pond 1, the total input of N was 364 kg ha^?1 and output was 359 kg ha^?1, whereas, in pond 2, the total input of N was 439 kg ha^?1 and total output was 331 kg ha^?1. The total input of P was 75 kg ha^?1 in pond 1 and 66 kg ha^?1 in pond 2, while total outputs for pond 1 and pond 2 were 74 and 65 kg ha^?1 respectively. Primary production from phytoplankton contributed the largest proportion of total OC (49–56%), while feed contributed the largest proportion of N (78–81%) and P (79–80%). Animals harvested from the aquaculture ponds accounted for the largest proportion of N (50–73%) and P (49–52%), and respiration accounted for the largest proportion of OC (43–61%) output from the system. The OC, N and P use efficiency of harvested animals was 30.30%, 70.19% and 50.14% in pond 1, respectively, and 21.03%, 46.95% and 46.47% in pond 2 respectively. In terms of nutrient use, the filter‐feeding bivalve, Sinonovacula constricta, was the most efficient species within the polyculture system.     

Effects of dietary inclusion of yacon,ginger and blueberry on growth,body composition and challenge test of juvenile rockfish (Sebastes schlegeli) against Edwardsiella tarda

Effects of dietary inclusion of yacon, Polymnia sonchifolia (YC), ginger, Zingiber officinale (GG), and blueberry, Vaccinium ashei (BB), on growth, body composition and challenge test of rockfish against Edwardsiella tarda compared to ethoxyquin were investigated. Three hundred and sixty fish were randomly distributed into 12 flow‐through tanks. Four experimental diets were prepared: the control diet (Con) with 0.1 g/kg ethoxyquin, and YC, GG and BB diets. Each diet was assigned to triplicate tanks of fish and hand‐fed for 8 weeks. Externally normal fish after fourth and eighth weeks of feeding trial were infected with Edwardsiella tarda for challenge test. Weight gain and specific growth rate (SGR) of fish fed the YC diet were greater than those of fish fed all other diets. Feed efficiency, protein efficiency ratio and protein retention of fish fed the YC diet were higher than those of fish fed all other diets. In the both fourth and eighth weeks of infection trials, mortality of fish fed the Con diet was higher than that of fish fed all other diets. In conclusion, dietary inclusion of YC, GG and BB increased weight gain and SGR of fish. YC, GG and BB for 4 and 8 weeks lowered mortality of fish at occurrence of E. tarda.     

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.     

Total organic carbon budget of integrated aquaculture system of sea cucumber Apostichopus japonicus,jellyfish Rhopilema esculenta and shrimp Fenneropenaeus chinensis

The total organic carbon (TOC) budget of the integrated aquaculture system of sea cucumber Apostichopus japonicus, jellyfish Rhopilema esculenta and shrimp Fenneropenaeus chinensis was investigated in a cofferdam (1.202 km²) for 13‐month field study. The results showed that the total input of TOC to the system was 498 300 kg year^?1, of which photosynthesis accounted for 26.8% of input TOC. The TOC production of the sea cucumber was 3668 kg year^?1, and that of the jellyfish and the shrimp was 714 kg year^?1. The TOC utilization rate of the input TOC by sea cucumber production was 7.4‰, and that of the integrated system increased by 18.9% than that of sea cucumber monoculture model through integration with jellyfish and shrimp. The outflow TOC from the system was 336 600 kg year^?1, which account for 92.7% of the inflow TOC. The integrated aquaculture system not only produce aquatic products, but also eliminate 7.3% of the inflow TOC, therefore, this system was not only a production system of aquatic products, but also an organic matter purification system for the coastal environment. The TOC utilization rate of the system can be probably improved by means of increasing stocking density of the sea cucumber and/or the shrimp.     

Prediction of cannibalism in juvenile black rockfish,Sebastes schlegelii (Hilgendorf, 1880), based on morphometric characteristics and paired trials

To predict the occurrence of cannibalism in the culture of larval black rockfish, Sebastes schlegelii, we developed a regression model based on morphometric measurements. First, 156 black rockfish were measured to obtain a theoretical model, that is L_{T prey} = 0.624 L_{T cannibal} + 0.835. Then, 202 trials of paired fish, together with 31 pairs of cannibalistic fish, were used to revise this model, and the practical model was constructed: L_{T prey} = 0.522 L_T cannibal + 4.908 (R² = 0.99). According to this equation, the threshold of size ratio was deduced, that is 1.30–1.69, which could be used to grade black rockfish by size and remove potential cannibals.