Assessment of dietary lecithin and cholesterol requirements of mud crab, Scylla serrata, megalopa using semi-purified microbound diets

The effects of varying dietary lecithin and cholesterol levels on growth, development and survival of mud crab, Scylla serrata, megalopa were evaluated using six semi‐purified, microbound diets formulated to be iso‐energetic and containing three levels of supplemental lecithin (0, 20 and 40 g kg⁻¹ diet dry weight) and two levels of supplemental cholesterol (0 and 7 g kg⁻¹ diet dry weight). Fifteen megalopa were reared individually in each treatment and the nutritional value of diets was assessed on basis of mean dry weight and mean carapace width of newly settled first crab stage, as well as development time to the first crab stage and overall survival. A significant interaction between supplemental dietary lecithin and supplemental dietary cholesterol was found for final mean dry weight of newly settled crabs, and highest survival (60%) was recorded for megalopa fed diets containing the highest levels of dietary lecithin (39.7–44.1 g kg⁻¹) (diet 5 and 6) regardless of whether diets were supplemented with cholesterol; this rate of survival was identical to that of megalopa fed live Artemia nauplii. The results indicate that supplemental dietary cholesterol may not be essential for mud crab megalopa when fed diets containing sufficient levels of supplemental dietary phospholipids.     

The effects of fish hydrolysate (CPSP) level on Octopus maya (Voss and Solis) diet: Digestive enzyme activity, blood metabolites, and energy balance

As has been demonstrated in previous studies, Octopus maya can be fed on artificial diets. In the present study six different diets were assayed. Five diets were designed to test the effect of percentage of inclusion of fish protein concentrate (CPSP: 0, 5, 10, 15, and 20%) and were offered to octopuses as a specifically designed artificial diet. The sixth diet consisted of frozen crab (Callinectes spp) and was used as control diet. Blood metabolites and energy budget of octopuses were evaluated to determine how CPSP levels modulate the digestive capacity and allow retaining energy for growth. Wild animals (316.4 ± 9.8 g) were used in the study. Results showed that CPSP produced a positive specific growth rate (SGR, % day^− 1) with high value in octopuses fed 15% CPSP level. A maximum growth rate of 0.86% day^− 1 was recorded in these animals, a value that is extremely low when compared with the SGR obtained when animals were fed fresh crab (3.7% day^− 1). In general, blood metabolites were affected by diet composition, indicating that some metabolites could reflect the nutritional and/or physiological status of octopus. Preliminary reference values for O. maya fed crab were found for glucose (0.09 ±0.02 mg/ml), lactate (0.004 ± 0.002 mg/ml), cholesterol (0.16 ± 0.02 mg/ml), acylglycerol (0.14 ± 0.01 mg/ml), protein (0.37 ± 0.04 mg/ml), hemocyanin (1.85 ± 0.04 mmol/l), and digestive gland glycogen (1.86 ± 0.3 mg/g). Total energy content can be used as an indicator of tissue metabolic reserves. In the present study, higher energy content in the digestive gland and muscle was observed in octopuses fed crab, followed by animals fed 15% CPSP. Results from the digestive gland indicated that the retained energy derived from glycogen, suggesting that lipids and protein were the main sources of variation linked with energy content. In general, digestive gland proteases activity and trypsin were induced in octopuses fed 15% CPSP. The capacity of O. maya juveniles to adjust their digestive enzymes to different types of food was evidenced. Essential amino acid content (EAA) of the diet was not a limiting factor. When dietary EAA profiles were compared with O. maya EAA profiles, all dietary EAA resulted in a higher concentration than whole body octopus composition. In the present study, all experimental groups ingested between 3300 and 4106 kJ wk^− 1 kg^− 1 without statistical differences among treatments, indicating that experimental diets were as attractive as crab. Differences were recorded in the proportion of absorbed energy (Ab, %) between CPSP-based and crab meat diets, suggesting digestion limitations associated with artificial diets. The present results indicate that the 15% CPSP diet had characteristics that stimulate digestive enzymes and reduce energetic costs associated with its digestion (HiE or SDA), channeling more biomass production than the other experimental diets.     

Successful culture of larvae of Litopenaeus vannamei fed a microbound formulated diet exclusively from either stage PZ2 or M1 to PL1

In three separate experiments, harpaticoid copepods Tisbe monozota (alive and dead) and a microparticulate microbound diet were evaluated as alternatives to live Artemia nauplii as food, beginning at either stage PZ2 or M1, in the larval culture of Litopenaeus vannamei. Larvae were cultured in 2 L round bottom flasks at a density of 150 L^− 1 (Experiment 1) and 100 L^− 1 ( 3.2 and 3.3) at 28 °C, 35‰ salinity and 12:12 LD photoperiod, and fed 4×/day^- 1. Larvae were initially fed a mixture of phytoplankton to stages PZ2 or M1 and then fed either live Artemia, live or dead copepods, or a microparticulate microbound diet. The experiments were terminated and all larvae were harvested when more than 80% of larvae had molted to postlarvae 1 (PL1) within any flask representing any of the treatments. The comparative value of the different diets and feeding regimes was determined by mean survival, mean dry weight and total length of individual larva, and percentage of surviving larvae that were PL1. Trypsin activity of samples of larvae from each treatment was also determined. The microparticulate microbound diet effectively served as a complete substitute for Artemia nauplii when fed beginning at stage M1. When fed at the beginning of the PZ2 stage, survival was comparable to that of larvae fed Artemia, but mean dry weight, mean total length, and percent of surviving larvae that were PL1 generally were significantly less. Responses to the feeding of copepods, whether fed dead or live, as a substitute were generally significantly less than those of larvae fed either the Artemia nauplii or the microparticulate diet. Values of trypsin activity (10^− 5 IU/μg^- 1 dry weight) corresponded to the relative proportions of the different larval stages within a treatment, with higher activity being characteristic of early stages. Previously demonstrated successful results with another species of crustacean suggest that the microparticulate microbound diet has characteristics that should be effective in the culture of the carnivorous stages of other crustacean and fish larvae that are currently fed live Artemia nauplii.     

Growth, survival and biochemical composition of spider crab Maja brachydactyla (Balss, 1922) (Decapoda: Majidae) larvae reared under different stocking densities, prey:larva ratios and diets

The spider crab Maja brachydactyla is overexploited on the NW coast of Spain. Aquaculture of this species can be the solution to the problem, and consequently, several attempts of intensive larval rearing have been conducted. However, most of the studies already published do not provide enough zoo technical data, especially in terms of larval and prey densities or the nutritional quality of diets used for rearing.Three experiments were carried out to evaluate the conditions for intensive larval rearing of M. brachydactyla. Larval stocking density (10, 50 and 100 larvae L^− 1), prey:larva ratio (15, 30 and 60 enriched Artemia larva^− 1) and diet (enriched Artemia, non-enriched Artemia and polychaete supplement) effects on growth and survival of this species were studied. For larval culture nine, 35 L, 150 μm mesh-bottomed PVC cylinders (triplicates for each treatment and larval stage) connected to a recirculation unit, were used. Temperature and salinity were kept constant at 18 °C and 36‰ respectively. A 12 to 18 day trial was conducted for each experiment and samples of larvae were collected at each larval stage (zoea I, zoea II, megalopa) in the inter-molt phase and at first juvenile. Survival, carapace length and width, dry weight (DW), and proximate biochemical content (protein, carbohydrates and total lipid) as well as lipid class composition were determined.Stocking densities of 100 larvae L^− 1 resulted in higher growth in DW and higher content in lipids and protein for zoea I (ZI) and zoea II (ZII) than 10 larvae L^− 1. However, survival decreased with increasing stocking density.The use of 60 preys larva^− 1 produced larvae with significantly higher DW and protein content, especially at ZII stage, than lower prey densities. Survival rate obtained feeding 60 preys larva^− 1 up to the megalopa stage was almost two-fold (42.2%) the rate obtained using 15 preys larva^− 1 (24.8%).Larvae fed on enriched Artemia (EA) showed an increase in weight up to megalopa (518.9 ± 26.5 μg) in contrast to larvae fed on non-enriched prey (A) (467.9 ± 6.9 μg). Variation in DW correlated with the total lipid content (L) of the larvae (L_EA = 70.1 ± 37.5 μg ind^− 1; L_A = 28.9 ± 3.2 μg ind^− 1) especially in terms of neutral lipids. The use of an initial density of at least 50 larvae L^− 1 and 60 enriched Artemia larva^− 1 can be considered the most adequate rearing parameters in order to obtain good results in growth and survival of M. brachydactyla.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Feeding experiments on the larvae of the fiddler crab Uca pugilator (Brachyura,Ocypodidae), reared in the laboratory

Larvae of Uca pugilator (Bosc) were reared in the laboratory from hatching to the megalopa stage on three different diets: (1) newly hatched Artemia salina nauplii (diet A), (2) the rotifer Brachionus plicatilis (O.F. Müller) and a ciliate Euplotes sp. (diet RC), and (3) a combination of the above two diets (diet ARC). The survival rate of zoeae fed diet A (90.0%) and diet ARC (93.8%) was much higher than that of the larvae fed diet RC (22.5%). The duration of the zoeal stages was significantly shorter for the larvae fed diet ARC than for those fed diets A and RC. The survival rate of megalopa larvae (reared on diets A and ARC in the zoeal stages) was high (above 90%) for megalopa fed Artemia nauplii only, as well as for those fed a combination of Artemia nauplii and shrimp. No significant differences in duration of the megalopa stage were found between the latter diet groups.     

Effect of different forms of Artemia biomass as a food source on survival,molting and growth rate of mud crab (Scylla paramamosain)

An experiment was conducted to evaluate the effect of different forms of Artemia biomass as a food source on survival, molting and growth rate of mud crab Scylla paramamosain. Instar 1 crablets with a mean weight of 0.0082 ± 0.0007 g were reared both individually and communally and fed with different diets consisting of fresh shrimp meat (control feed), live Artemia biomass, frozen Artemia biomass and a dried Artemia‐based formulated feed for 40 days. The highest survival was obtained for crablets receiving live Artemia (92.5% and 75.8%) followed by the groups fed with frozen biomass (90.0% and 47.5%), the control feed (72.5% and 24.2%) and the dried Artemia‐based diet (60.0% and 21.7%) for individual and communal cultures, respectively. The intermolt period, the total number of moltings and the growth rate, which were determined on individually reared crabs, showed the same pattern as for survival. The results suggest that crab performance decreased in the following order: live Artemia>frozen Artemia > fresh shrimp meat > dried Artemia‐based formulated feed. Live Artemia biomass proved an ideal feed for nursery of Scylla paramamosain crabs. Frozen Artemia biomass may be an alternative in times of shortage. Our findings illustrate the high potential for local utilization of Artemia biomass in Vietnam for reliable production of mud crab juveniles.     

The effects of algal diets on population growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis

As natural diets of fish larvae, a number of calanoid copepod species are being investigated for use as live prey in aquaculture hatcheries. One of these, the tropical calanoid copepod, Acartia sinjiensis, has good potential as a live feed for tropical reef fish larvae. However, the rearing techniques for A. sinjiensis require further development to improve productivity. This study was carried out to investigate the population growth and egg hatching success of A. sinjiensis when fed a range of mono-and binary algal diets, including algae in the form of frozen paste.For the population growth experiment, the final A. sinjiensis population, including eggs, nauplii, copepodites and adults, was determined after feeding eight algal diets (two frozen algae, four live monoalgal and two live binary algal diets) for 8 days at temperature 28 ± 1 °C; salinity 34 ± 1 psu and photoperiod 12 L:12 D. Five replicates, with an initial 12 adult A. sinjiensis per replicate, were set up for each treatment. In a separate experiment, effects of diets on egg hatching success were examined after 48 h incubation of eggs produced by A. sinjiensis fed the same eight diets.The results showed that diet significantly affected both population growth and hatching success of A. sinjiensis. Of the diets tested, the binary algal diets were more successful than monoalgal diets, while the frozen algae had little dietary value. The highest population growth was recorded on A. sinjiensis fed a binary diet of Tetraselmis chuii and the Tahitian strain of Isochrysis sp. (T-ISO) (final population: 1091 ± 80), which was significantly higher (P < 0.001) than all other diets tested except for the other binary diet of Nannochloropsis sp. and T-ISO (final population: 897 ± 123). Diet also had a significant effect (P < 0.001) on egg hatching rate, though the highest hatch rate was recorded with eggs produced by A. sinjiensis fed binary diet Nannochloropsis sp. and T-ISO (88.1 ± 2.1%), this was not significantly different from that of eggs produced by A. sinjiensis fed either T-ISO alone (88.0 ± 1.7%) or the binary algal diet of T. chuii and T-ISO (76.4 ± 7.1%). The results of this study suggest that among the diets tested, a combination of live T. chuii and T-ISO was the best for the culture of A. sinjiensis.     

Effects of water quality,antibiotics, phytoplankton and food on survival and development of larvae of Scylla serrata (Crustacea: Portunidae)

Larvae of the portunid crab Scylla serrata were successfully reared using a combination of antibiotics (penicillin-G + polymyxin-B), phytoplankton (Chlorella sp.) and appropriate food (Artemia salina nauplii).Antibiotics enhanced premetamorphic survival of zoeae while leaving rate of zoeal development and success of metamorphosis to megalopa unaltered. Water filtration and ultraviolet sterilization had no significant effect on rates of zoeal survival or development. Although antibiotics did not affect metamorphosis of megalopa to the crab stage, the antibiotic mixture may have been detrimental to survival of megalopae.The presence of Chlorella, unlike antibiotics, left zoeal survival unaffected while stimulating production of megalopae. As with antibiotics, time from hatching to megalopa formation was unaltered by the presence of Chlorella.Survival and development of zoeae varied with type and concentration of food organisms used. Feeding with Artemia nauplii produced highest larval survival and continued development. Rotifers and zooplankton as a food source failed to support zoeal survival to the onset of metamorphosis and failed to enhance survival when fed in combination with Artemia nauplii.A daily optimum food concentration of ten Artemia nauplii per ml was established for zoeal survival, while success of metamorphosis of zoeae to megalopa increased as food concentration increased from 5 to 16 nauplii per ml. Rate of development was comparable in all zoeal feeding experiments.Megalopae survived better at the higher food (Artemia) concentrations used and tended to develop more rapidly to the crab stage at higher food levels.     

Suitability of the copepod, Acartia clausi as a live feed for Seabass larvae (Lates calcarifer Bloch): Compared to traditional live-food organisms with special emphasis on the nutritional value

Though artificial propagation of Asian seabass Lates calcarifer (Bloch) in captivity through induced breeding techniques is standardized under Indian conditions, larval and nursery rearing techniques including suitable nursery feeds have to be standardized to obtain better survival and growth. Feeding experiments in triplicate were conducted to evaluate the suitability of the marine copepod Acartia clausi as live prey for fourteen day-old seabass larvae (6.53 ± 0.06 mm; 8.58 ± 0.33 mg) and compared with the traditional live prey, rotifers and Artemia nauplii. While A. clausi and rotifers were mass produced using algae Isochrysis galbana, Chaetoceros affinis and Chlorella marina, Artemia nauplii were produced using cysts. Nutritional quality of cultured copepods was evaluated based on the proximate composition, amino acid and fatty acid composition, and compared with that of rotifers and Artemia nauplii. Proximate composition varied significantly (P < 0.05) among the different live feeds. A. clausi showed higher protein (63.12%) and lipid (16.65%) content than Artemia nauplii and rotifers. Total essential amino acids content was 2% lower in A. clausi compared to that in Artemia nauplii. Fatty acid profiles of the live feed organisms showed that A. clausi is a rich source of n − 3 fatty acids. The total n − 3 fatty acid content of A. clausi was 33.94%. Length, weight overall weight gain and survivorship were significantly (P < 0.05) different among the dietary treatments, and weight gain was comparatively higher in A. clausi fed larvae. Survival of seabass larvae fed A. clausi was obtained highest as 58.13% against the lower values of 39.93% and 41.62% in larvae fed rotifer and Artemia nauplii respectively. Final carcass composition of the larvae of L. calcarifer fed different live-food organisms showed significant differences (P < 0.05) among the dietary treatments. The fatty acid composition of the dietary treatments was reflected to a certain extent in the fatty acid composition of the seabass larvae. The present investigation revealed the nutritional value of calanoid copepod and thus underlining its usefulness as a suitable live-food organism for rearing larvae of the commercially valuable Asian seabass.     

Effect of dietary supplementation of phospholipids and highly unsaturated fatty acids on reproductive performance and offspring quality of Chinese mitten crab, Eriocheir sinensis (H. Milne-Edwards), female broodstock

This study was conducted to determine the optimum phospholipids: highly unsaturated fatty acids (PL/HUFA) ratio in maturation diets for female Chinese mitten crab, Eriocheir sinensis broodstock. Five isolipidtic and isonitrogenous diets were formulated to contain different amounts of pork lard, fish oil and soy lecithin with resulting PL/HUFA levels (%dry weight) of 1.4/0.3 (Diet 1), 1.6/2.5 (Diet 2), 2.3/2.0 (Diet 3), 3.8/1.5 (Diet 4) and 5.1/0.8 (Diet 5). Diet 1 lacked supplemental fish oil and soy lecithin and served as the control. Each of the five formulated diets was fed to a total of 150 female crabs for a period of 7 months. All diet treatments were triplicated with 50 crabs stocked in a pond as a replicate. After 6 months, male crabs were introduced into the ponds where females were kept, mating, spawning and egg hatching (in seawater) occurred in the following month. The nutritional value of various formulated diets was assessed based on survival, gonadosomatic index (GSI), egg production per female, fecundity (eggs/g female weight) of the females fed those diets and egg and larval (newly hatched zoea I larvae) quality.Females fed Diet 1 had the lowest GSI, egg production per female and fecundity while no significant difference were found for survival, hepatosomatic index (HSI) and the percentage of female spawned among the diet treatments (P > 0.05). Although there were no significant differences in egg diameter, egg wet or dry weights among all treatments (P > 0.05), the highest and the second highest proportions of essential fatty acids (EFA), i.e. 20:5n-3 (EPA), 22:6n-3 (DHA) and HUFA were found in the eggs produced by the females fed Diet 3 and Diet 2, respectively. Statistical analysis showed that EPA, DHA and HUFA in eggs produced by females fed Diet 3 were significantly higher than those from the other treatments (P < 0.05). Meanwhile, zoea I larvae from crabs fed Diet 3 had significantly larger carapace length than those from the other four treatments (P < 0.05). The zoea I larvae from crabs fed Diet 2 and Diet 3 also showed generally better tolerance to starvation and osmotic shock.In conclusion, our results indicated that diet included levels of PL/HUFA around 1.6/2.5 or 2.3/2.0 (% dry weight) fed female E. sinensis broodstock supported increased fecundity and elevated level of HUFA in egg, which in turn resulted in improved overall quality of newly hatched larvae.     

Performance of different types of diets on experimental larval rearing of endangered Chitala chitala (Hamilton) in recirculatory system

This is the first report on the successful larval rearing of captive bred population of Chitala chitala (Hamilton). C. chitala is one of the endangered fresh water fish species in India for which the development of controlled larval rearing procedures are needed for stock enhancement. Fifteen days old post-hatchlings were stocked for 28 d in a 30 L recirculatory tanks using eight different diets i.e. live feed (tubifex worms, chironomous larvae, zooplanktons,), dry feed (dry tubifex, spirulina, daphnia) and other non-conventional feed (fish eggs and boiled egg-yolk). Fishes accepted all types of diets. The study revealed that specific growth rate (SGR) was higher in post-hatchlings fed on live tubifex worms (2.40 ± 0.72) followed by fish eggs (2.15 ± 0.71), dry tubifex (2.12 ± 0.40), chironomous larvae (1.91 ± 0.44), spirulina (1.79 ± 0.38), daphnia (1.42 ± 0.79) and planktons (1.37 ± 0.77) whereas minimum SGR was recorded with boiled egg-yolk (0.63 ± 0.5). A highly significant difference (p < 0.01) in SGR was observed in fish fed on live feed (tubifex worms, chironomous larvae, planktons, spirulina), dry tubifex and fish eggs whereas for daphnia and boiled egg-yolk it was only significant (p < 0.05). The final mean weight and weight gain showed highly significant difference (p < 0.01) in live tubifex, zooplanktons, spirulina, chironomous larvae, dry tubifex and fish eggs, whereas daphnia and boiled egg-yolk fed larvae showed significant difference (p < 0.05). Highest mean survival rate on day 28 was observed in live tubifex worms (94%) and chironomous larvae (92%). The post-hatchlings reared with spirulina and daphnia showed same survival rate of 88% whereas the lowest mean survival of 66% was recorded in boiled egg-yolk. The experiments showed that captive bred post-hatchlings of C.chitala could be reared in experimental recirculatory system for attaining higher growth and survival during early life stages. However, methods to improve the larval rearing have to be improved further for commercial farming of the species.     

Variations on development and stress defences in Solea senegalensis larvae fed on live and microencapsulated diets

Growth, development, antioxidant enzymes, stress proteins (HSP70 and HSP60), lipid peroxidation (LP) and histology in Solea senegalensis larvae were followed from 8 to 30 days post hatching (dph). Larvae were fed on three different diets: (1) live Artemia nauplii, (2) microcapsules elaborated by internal gelation, (MA) and (3) these same microcapsules but 10-fold supplemented with vitamin A (MAV). The Artemia fed group showed higher growth and a faster metamorphosis than the ones fed with microencapsulated diets, although all had similarly high survival rates of 80%. Vitamin A (VA) supplementation improved growth and development from 15 dph in relation to the strictly inert diet (MA). Larvae fed with Artemia showed organs and tissues with a normal pattern of development, whereas histological alterations were seen in larvae fed with both inert diets. The antioxidant enzymes: catalase (KAT), superoxide dismutase (SOD), and total glutathione peroxidase (t-GPX) as well as LP levels and stress proteins (HSP70 but not HSP60), measured in whole larvae, showed diet and age dependence in their response. Larvae fed with both inert diets showed similar biomarker activities, but these activities were different (p < 0.05) from larvae fed with Artemia. That is, KAT and HSP70 were lower in larvae fed with live prey and t-GPX and LP levels were lower in larvae fed with the inert food. Among the factors responsible for increased antioxidant defenses were the initiation of metamorphosis and the use of inert food. This study suggests the usefulness of the biomarkers selected as tools to evaluate the effects of compound diets on larvae.     

Effect of dietary cholesterol levels on growth performance,body composition and gene expression of juvenile mud crab Scylla paramamosain

A 56‐day feeding trial was conducted to investigate the effect of dietary cholesterol (CHOL) levels on growth performance, body composition and gene expression of juvenile mud crab (Scylla paramamosain). Four isonitrogenous and isoenergetic diets were formulated with 0.4%, 0.8%, 1.2% and 1.6% CHOL supplemented, and the final dietary CHOL concentrations were 0.72%, 1.11%, 1.49% and 1.83% respectively. Each dietary treatment was performed with three replicates (28 crabs per replicate, initial body weight 0.04 g). At the termination of the experiment, although the survival had no statistic difference in all treatments, the mud crabs fed the lowest CHOL diet had the lowest survival rate. The weight gain (WG) of mud crab significantly increased with dietary CHOL level up to 1.11% and then significantly decreased with dietary CHOL level further increased. The total‐cholesterol (T‐CHOL) content in whole body had an increasing trend with the dietary CHOL level increased. Besides, dietary CHOL supplement generally increased the hepatic superoxide dismutase (SOD) activity, and the mud crabs fed diets CHOL1.11 and CHOL1.49 showed significantly higher value than those fed other diets. The hepatic aspartate aminotransferase (AST) activity decreased slightly with dietary CHOL level up to 1.11% and then significantly increased with CHOL level further increased. The mRNA expression of ecdysone receptor (EcR) gene in the eyestalk obviously increased with dietary CHOL level up to 1.11% and then significantly decreased with dietary CHOL further increased. These results suggested that about 1.11% dietary CHOL seem fulfil to maintain good growth performance and healthy condition for juvenile S. paramamosain.     

Effects of four natural diets on the culture performance and biochemical composition of megalopa of Eriocheir sinensis during desalination period

Frozen copepods, frozen adult Artemia, live rotifers and live Artemia nauplii were used as feed for the megalopa of Eriocheir sinensis during desalination stage to determine the most nutritious and cost‐effective feed. The biochemical composition of the four feeds was analysed. Then, the individual wet weight, individual dry weight, survival rate, the cost and the biochemical composition of the megalopa were measured after the desalination. The resulting effects on the metamorphosis ratio and the duration of the megalopa were also assessed. The results showed that the duration of desalinated megalopa to first stage crab for all treatments was approximately 3.5 days, while the metamorphosis rate of the copepod treatment was significantly higher than the other treatments with the value of 96.33 ± 3.79%. In terms of biochemical composition, after desalinization, the moisture content of megalopa in the adult Artemia treatment was the highest, and the total lipid in the Artemia nauplii treatment and rotifer treatment was significantly higher than the other two treatments. The desalinated megalopa from copepod treatment had the highest percentages of DHA (22:6n‐3) and EPA (20:5n‐3) among the four treatments. The desalinated megalopa from copepod treatment and rotifer treatment had the higher contents of protein. In conclusion, frozen copepods have higher nutritional value for the megalopa during the desalination period. Therefore, feeding the megalopa with copepods can improve the culture performance and megalopa quality, and can also reduce the feed cost during desalination stage.     

Preliminary dataon larval survival of South American catfish,Rhamdia sapo

Larvae of Rhamdia sapo were fed four test diets of Artemia nauplii; a balanced diet formula; boiled egg yolk, coagulated blood and liver homogenate; and live zooplankton. The homogenized mixture was the most effective for growth. Highest survival was recorded with the Artemia diet. Larger trials performed in indoor raceways, using the homogenate diet for 10 days followed by a balanced ration, were successful before transfer to grow-out ponds.     

First feeding of Octopus vulgaris Cuvier, 1797 paralarvae using Artemia: Effect of prey size, prey density and feeding frequency

Different assays related to the first feeding of Octopus vulgaris Cuvier, 1797 are compiled in this paper. They include: age at initial feeding age, prey size selection and optimal density, attack timing after feeding, and effect of dose number on the number of captures. Prey capture and ingestion processes were also analysed. Food supplied was cultured Artemia sp. Each assay lasted 15 min.Although paralarvae already start to feed on the hatching day (day 0), it is during day 2 when a greater number of attacks is recorded (81.7 ± 14.7% paralarvae attack). They mainly prefer (significance level α = 0.05) large Artemia, 1.4 ± 0.4 mm (77.0 ± 5.6% of the total attacks) than small Artemia, 0.8 ± 0.1 mm (23.0 ± 5.6%). There is also a slight predilection for the lowest Artemia concentration (33.3 ±12.6% paralarvae attack in a 0.1 Artemia ml^− 1 density, opposite 16.7 ± 7.6 and 18.3 ± 7.6% in densities of 0.5 and 1 Artemia ml^− 1 respectively). The greatest predatory activity is recorded during the first 5 min after food is supplied (72.2 ± 25.5%). An increase in the predatory activity was also observed when food was distributed in several doses instead of a single dose (75.0 ± 10.0% and 46.7 ± 17.6% respectively). It was proved for the first time that paralarvae completely ingest their preys (including their exoskeletons), in this case Artemia. Time needed for their total ingestion ranges between 4 and 10 min.     

Growth performance,survival, feed utilization and nutrient utilization of African catfish (Clarias gariepinus) larvae co‐fed Artemia and a micro‐diet containing freshwater atyid shrimp (Caridina nilotica) during weaning

Problems of limited number of dry feeds as supplement or replacement of live feeds have led to poor larval nutrition in many species of fish. Therefore, the suitability of co‐feeding 8‐day‐old African catfish (Clarias gariepinus) posthatch larvae using live feed (Artemia salina) and formulated dry diet containing freshwater atyid shrimp (Caridina nilotica) during weaning was investigated. The experiment ended after 21 days of culture and respective groups compared on the basis of growth performance, survival, feed utilization and nutrient utilization. Larvae co‐fed using 50%Artemia and 50% formulated dry diet resulted in significantly (P < 0.05) better growth performance, food gain ratio (FGR), protein efficiency ratio (PER) and productive protein values (PPV) than other treatments. The lowest growth performance occurred in larvae weaned using 100% formulated and commercial dry diets. Better survival of over 90% was obtained in larvae weaned using 50%Artemia and 50% dry diet, while abrupt weaning using 100% dry diets resulted in lower survival (<75%). These results support a recommendation of co‐feeding C. gariepinus larvae using a formulated dry diet containing C. nilotica and 50% live feed when weaning is performed after 8 days posthatching period.     

Dietary mannan oligosaccharide enhances salinity tolerance and gut development of larval cobia

The potential beneficial effects of supplementing live feeds with mannan oligosaccharide (MOS; BioMos®) upon cobia Rachycentron canadum larval performance were examined. Characteristics of fish examined included survival to weaning, growth, ability to withstand osmotic stress and the degree of development of the brush border of the intestine. Live feeds included rotifers (Brachionus plicatilis) and Artemia which were enriched for 24 h with a commercial enrichment media alone or in combination with 0.2% (dry weight basis) MOS. Salinity challenges were performed at 6 days post-hatch (dph) and at 7, 13, and 14 dph (0 and 65 g L^− 1 for 6 dph; 0 and 55^− 1 7+ dph) corresponding to transitions in feeding, to examine the ability of larval cobia to survive stress. Differences (P < 0.05) in survival, favoring cobia receiving MOS-supplemented feeds were discerned at 6 and 7 days post-hatch (dph) when fish were challenged at 0 g L^− 1 and at 13 dph when challenged with 55 g L^− 1 salinity water. Electron microscopy of the mid-intestine of developing larvae revealed that MOS-supplemented diets enhanced (P < 0.05) the height of microvilli while reducing (P < 0.05) the occurrence and size of supranuclear vacuoles. Supplementation of diets with MOS could assist cobia larvae in maintaining allostasis especially when reared at sub-optimal salinities.     

Development of larval culture techniques for the shore crab, <Emphasis Type="Italic">Carcinus maenas</Emphasis> (L)

The shore crab Carcinus maenas is a commercially important species, utilised as sea angling bait as well as supporting a European-wide fishery. Hatchery production could provide an alternative source of bait crabs, alleviating potential competition between these sectors and environmental concerns regarding bait collection practices. A series of experiments were carried to investigate the potential for hatchery production, focusing on effects of dietary regimes and stocking densities through the zoeal stages and the influence of tank substrates and stocking density during the megalopa stage. Inclusion of the rotifer Brachionus plicatilis as live food for early larval stages conveyed no advantage in terms of survival or rate of development compared to a diet of Artemia nauplii. Increasing zoea stocking densities (from 94 to 557 l⁻¹) had a significantly negative effect upon survival to the megalopa stage (from 75% down to 47%), although this was off-set by a significant increase in production, with 260 megalopae⁻¹ produced from an initial density of 557 zoeae l⁻¹. The inclusion of substrates for megalopa stages had no impact on production or development rate, compared to tanks with no substrate. The completely benthic behaviour of megalopae indicates that tank floor area will be a limiting factor for crab production. Increasing stocking density of megalopae was found to significantly and negatively influence survival, although above 10,000 megalopae m⁻² the rate of decline in survival stabilised and maximum production (3,114 juveniles m⁻²) of juvenile crabs could be achieved at the highest stocking densities tested (40,000 m⁻²).