Enrichment of Rotifers Brachionus plicatilis with Eicosapentaenoic Acid and Docosahexaenoic Acid Produced by Bacteria

Abstract— Two bacterial strains, rich in either eicosapentaenoic acid [EPA, 20:5(n-3)] ( Shewanella gel-idimarina ACAM 456) or docosahexaenoic acid [DHA, 22:6(n-3)] ( Colwellia psychroeryrhrus ACAM 605) were tested for their ability to enrich rotifers Erachionus plicatilis in these polyunsaturated fatty acids. Rotifers were exposed for 24 h to each bacterial strain and to a mixture of the two strains. They were then harvested and their fatty acid compositions were analysed and compared to those of rotifers that had been either starved or fed yeast Saccharomyces cerevisiae or microalgae Tetraselmis suecica in 2-L glass flasks. Exposure to 1.4 × 10⁹ cells/ml of the EPA-producing bacterium only resulted in rotifer EPA levels increasing from 0.1% to 1.2% of total dry weight (%dw). Similarly, following exposure to 1.0 × 10⁹ cells/mL of the DHA-producing bacterium only, rotifer DHA levels increased from below detection to 0.1% dw. When exposed to a mixture of the two bacterial strains, containing 7.0 × 10⁸ celldml of the EPA producer and 5.0 × 10⁸ cells/mL of the DHA producer, the rotifers'final EPA and DHA levels were 0.5% dw and 0.3% dw respectively. Although feeding strategies need refining, these results show, for the first time, that rotifers can be enriched with DHA from bacteria, and that rotifers can be enriched simultaneously with both DHA and EPA from different bacterial strains.     

Effect of dietary lipid level on muscle composition in Atlantic salmon Salmo salar

This study was conducted to determine the effects of feeding increasing lipid concentrations (310, 380 and 470 g kg^–1 lipid on dry weight) in diets based mainly on herring byproducts to Atlantic salmon Salmo salar . The diets were isonitrogenous, varying in dietary lipid content at the expense dietary starch. Average fish weight increased from 1.2 kg in April to 2.2–2.7 kg at the end of the feeding trial in September. Significantly greater growth was found in fish fed either the 380 g kg⁻¹ or the 470 g kg⁻¹ lipid diets compared with the 310 g kg⁻¹ lipid diet. Muscle lipid content increased in all dietary groups on a wet weight basis from 7.7 ± 1.4% to 12 ± 3% in salmon fed the 310 g kg⁻¹ lipid diet, and to 16 ± 2% in salmon fed the 380 g kg⁻¹ and 470 g kg⁻¹ lipid diets. In fish of similar weight there was a positive correlation between dietary lipid and muscle lipid concentrations. Low concentrations of muscle glycogen were detected in fish fed each of the diets, while muscle vitamin E concentrations slowly decreased as muscle lipid increased. Muscle fatty acid composition reflected dietary fatty acid profiles, containing similar percentages of total saturated, monoenic and n-3 fatty acids (20:5n-3 and 22:6n-3) in fish from all dietary treatment groups. However, a higher ratio of n-3/n-6 was found in muscle from fish fed the 470 g kg⁻¹ lipid diet compared with the other two groups. Blood chemistry values varied somewhat, but all values were within normal ranges for Atlantic salmon of these sizes.     

Temporal variation of seasonality of egg production and the spawning biomass of Pacific anchovy, Engraulis japonicus, in the southern waters of Korea in 1983–1994

Embryonic mortality, egg production and the spawning stock biomass of Pacific anchovy, Engraulis japonicus , off Southern Korea during 1983–1994, and their biological response to oceanographic features in spring and summer, were analysed. The instantaneous mortality rate (IMR) of embryonic stages decreased in spring and increased in summer, with a range of 0.33–1.23 day^–1 in spring and 0.78–1.69 day^–1 in summer. Egg production in summer was three times that during spring and production was low in the late 1980s. Mean lengths of yolk-sac larvae and adult females were greater in spring than in summer, whereas spawning fraction and spawning stock ratio (spawning biomass:adult biomass) were lower in spring than summer. Estimated mean spawning stock biomass ranged from 141 × 10³ to 380 × 10³ MT in spring and from 221 × 10³ to 557 × 10³ MT in summer. Statistically, the seasonal and long-term trends of embryonic mortality, egg production and spawning stock biomass of Pacific anchovy can be explained largely by spring warming, summer cooling and by less abundant zooplankton in the late 1980s.     

Effects of processing on physical characteristics of diets with distinct levels of carbohydrates and lipids: the outcomes on the growth of pacu (Piaractus mesopotamicus)

The use efficiency and feed conversion of extruded and pelletized diets were compared. Eight isoproteic diets (220 g kg⁻¹ digestible protein) were assayed for 90 days in a 2 × 2 × 2 multifactorial design with two carbohydrate levels (400 and 500 g kg⁻¹), two lipids levels (40 and 80 g kg⁻¹) and two diet processing (pelletization and extrusion) with three repetitions. The growth of Piaractus mesopotamicus fed with these diets and the quality control indices of diets were gauged. The density of extruded diets was lower as carbohydrate level was 400 g kg⁻¹ and lipid 40 g kg⁻¹. The interaction carbohydrate and diet processing presented higher leaching value for low carbohydrate level in extruded diet. Fish fed with extruded diets presented the best feed conversion and protein efficiency ratio. When high levels of carbohydrate and lipid are combined, the weight gain is impaired. The interaction between diet processing diet and lipid levels resulted in the best fish performance when pelletized diets with 40 g kg⁻¹ lipid or extruded diets with 80 g kg⁻¹ lipid were considered. The protein efficiency ratio increased with the increment of carbohydrates in the pelletized diets. The fish show low tolerance to lipids and a preference for carbohydrate when the lipid productive values are taken into account.     

Natural Spawning of Captive Bigeye Scad Selar crumenophthalmus in Hawaii

Abstract.— Juvenile bigeye scad Selar crumenophthalmus were caught from the wild by hook and line and reared over 3 yr to establish a spawning broodstock of this species. The size at first maturity for captive males and females was 19-cm and 25-cm fork length (FL). Natural spawning of captive individuals occurred during the first year in captivity. The broodstock were observed to spawn repeatedly throughout their second and third years in culture. This species is a multiple spawner producing approximately 96 × 10³ to 121 × 10³ eggs/spawning group, 5 to 10 times per year. Stocking densities of 1–8 individuals/m³ were found to be suitable for natural spawning to occur. Collectively, the results indicate that this species exhibits notable potential for artificial propagation.     

Effects of dietary organic acids on growth, nutrient digestibility and gut microflora of red hybrid tilapia, Oreochromis sp., and subsequent survival during a challenge test with Streptococcus agalactiae

A 14-week feeding trial was conducted to determine the effects of dietary organic acids. The experimental diets were added with 0, 1, 2 or 3 g kg⁻¹ of a novel organic acid blend or with 2 g kg⁻¹ of potassium diformate and fed to triplicate groups of red hybrid tilapia ( Oreochromis sp.). Upon completion, tilapia were challenged by immersion with Streptococcus agalactiae . There was no significant difference ( P >0.05) in the growth, feed utilization and nutrient digestibility among treatment groups despite a trend towards improved results with fish fed organic acid-supplemented diets. Diet pH decreased, causing a reduction in the digesta pH of the stomach and gut. Total bacteria per gram of faeces were significantly ( P <0.05) reduced from 1.81 × 10⁸ colony-forming units (CFU) (control group) up to 0.67 × 10⁸ CFU in the fish fed organic acid diets. A similar trend was observed for adherent gut bacteria. Cumulative mortality of fish fed no organic acids was higher compared with fish fed organic acid-supplemented diets at 16 days post challenge. The data showed that dietary organic acids can exert strong anti-microbial effects and have the potential to exert beneficial effects on growth, nutrient utilization and disease resistance in tilapia.     

Effects of dietary phosphorus and lipid levels on utilization and excretion of phosphorus and nitrogen by rainbow trout (Oncorhynchus mykiss). 1. Laboratory-scale study

Nutritional strategies to reduce both phosphorus (P) and nitrogen (N) excretion relative to growth of rainbow trout were tested in a 2 × 3 factorial experiment. The two factors were `dietary P level' and `dietary lipid level.' Reduction in dietary P from 14 to 8 g kg^–1 dry diet was achieved by partial substitution of dietary fish meal with a combination of full-fat soyabean meal, corn gluten and spray-dried blood meal. Triplicate tanks of 35 rainbow trout per tank were fed experimental diets for 16 weeks and grew from approximately 40 to 250 g, in 15 °C spring water. All tanks were fed the same percent biomass per day. Diets were isonitrogenous, and dietary energy varied with dietary lipid. Diet digestibility data and results of the experiment were used to construct N and P budgets for the fish fed the various diets. A reduction in dietary fish meal from 500 to 200 g kg^–1 dry diet, corresponding to a reduction in dietary P from 14 to 8 g kg^–1 dry diet, resulted in >50% reductions in both solid and dissolved P waste, but did not affect growth, feed efficiency ratio (FER) or sensory characteristics of rainbow trout. Increasing dietary lipid from 170 to 310 g kg^–1 dry diet led to higher growth rate and FER, and lower total N waste relative to weight gain, but did not change protein retention. Increasing dietary lipid level increased deposition of lipid in whole bodies of rainbow trout, and resulted in discernible differences in sensory characteristics of trout fillets.     

Natural abundance of 15N and 13C in fish tissues and the use of stable isotopes as dietary protein tracers in rainbow trout and gilthead sea bream

For developing efficient diets, two sets of experiments examined whether the use and allocation of dietary protein can be traced by labelling with stable isotopes (¹⁵N and ¹³C) in two culture fish ( Oncorhynchus mykiss and Sparus aurata) . In the first experiment, natural abundance and tissue distribution of these isotopes were determined, by measuring the δ¹³C and δ¹⁵N values by isotopic ratio mass spectrometry, in fingerlings (14–17 g) adapted to diets differing in the percentage of fish meal replacement by plant protein sources. For both species, δ¹⁵N and δ¹³C were greater in tissues with higher protein and lower lipid content. Delta ¹⁵N of diets and tissues decreased as replacement increased, suggesting δ¹⁵N can be used as a marker for dietary protein origin. The ¹⁵N fractionation (δ¹⁵N fish − δ¹⁵N diet) differed between groups, and could thus be used to indicate protein catabolism. In the second experiment, fish (75–90 g) of each species ingested a diet enriched with ¹⁵N-protein (10 g kg⁻¹ diet) and ¹³C-protein (30 g kg⁻¹ diet). These proportions were suitable for determining that the delta values of tissue components were high enough above natural levels to allow protein allocation to be traced at 11 and 24 h after feeding, and revealed clear metabolic differences between species.     

Effects of high dose of vitamin A on reproduction and egg quality of Japanese flounder Paralichthys olivaceus

ABSTRACT: The present study was conducted to investigate the effects of dietary vitamin A on reproduction and egg quality in Japanese flounder Paralichthys olivaceus . Broodstock were fed experimental pellets containing two levels of vitamin A [11 × 10³ IU/100 g (control diet; CD), 337 × 10³ IU/100 g (experimental diet; ED)] for approximately 2 months before spawning and during the spawning period. Two groups of five females (average weight 1.4 kg) and 10 males (average weight 0.7 kg) were randomly allocated to two 30 m³ indoor tanks. Total egg production of the CD group was slightly higher than the ED group. Percentage of buoyant eggs and hatching rate of the ED group were significantly higher than the CD. In other egg quality parameters, such as percentage abnormal larvae and starvation tolerance of larvae, no notable difference was found between these two groups. At the end of the experiment, the skin color of broodstock in the ED group was darker than that of the CD group. Vitamin A content in eggs of the ED group was significantly higher than that of the CD group. However, the difference in vitamin A content in eggs between the ED and CD groups was much smaller than that in the liver of the females between the two groups. These results indicate that feeding broodstock a higher level of vitamin A increases the vitamin A content in eggs but does not affect egg quality in Japanese flounder because excess dietary vitamin A was stored mainly in the broodstocks' liver.     

Effects of the probiotic, Lactobacillus acidophilus, on the growth performance, haematology parameters and immunoglobulin concentration in African Catfish (Clarias gariepinus, Burchell 1822) fingerling

This experiment was carried out to evaluate the effects of the probiotic, Lactobacillus acidophilus , on the growth performance, haematology parameters and immunoglobulin concentration in African catfish Clarias gariepinus fingerling. Two experimental diets were formulated to contain 35 g kg⁻¹ crude protein and 10 g kg⁻¹ lipids accordingly and fed three times daily for 12 weeks to 25 C. gariepinus fingerlings per fibreglass tank in 12 replicates each. The control diet was prepared with no probiotic supplementation whereas the second diet was prepared supplemented with a probiotic, L. acidophilus , containing about 3.01 × 10⁷ colonies/g of diet. The results show that growth performance [specific growth rate (SGR) and relative growth rate (RGR)], nutrient utilization [protein efficiency ratio (PER) and feed conversion ratio (FCR)] and survival were significantly ( P <0.05) higher in fish maintained on the probiotic-supplemented diet compared with those on the control diet. Haematology parameters (packed cell volume, haemoglobin, erythrocyte sedimentation rate, red blood cell and white blood cell, total serum protein, Ca²⁺, Mg²⁺, Cl⁻, glucose and cholesterol) and total immunoglobulin concentrations were also significantly better in fish fed the probiotic-supplemented diet than in the control. Although the water quality parameters monitored were better in the fish fed the probiotic-supplemented diet than in the control, the parameters were not significantly different ( P >0.05). From the results of this experiment, we conclude that L. acidophilus can be used as a probiotic agent in African catfish culture, to enhance fish health, survival and better feed efficiency and growth performance.     

Fertilization efficiency of cryopreserved sperm from striped catfish, Pangasius hypophthalmus (Sauvage)

The fertilization efficiency of cryopreserved sperm was compared with fresh sperm from striped catfish, Pangasius hypophthalmus . Of the two sets of experiments carried out, the first compared four sperm doses using fresh sperm and fresh eggs. The second experiment compared six concentrations of cryopreserved sperm ranging from 6.94 × 10⁷ to 6.94 × 10¹⁰ to fertilize 100 eggs per batch. Fertilization, hatch and survival rates were compared between cryopreserved and fresh sperm. The highest fertilization rate (53.75±1.62%) was achieved with a sperm dose of 6.94 × 10⁸. Increasing the sperm dose to 3.47 × 10⁹ did not increase the fertilization rate, indicating that the optimum sperm:egg ratio lies between 6.94 × 10⁶ and 3.47 × 10⁷ sperm per egg. Both highest (6.94 × 10¹⁰) and the lowest (6.94 × 10⁷) sperm doses resulted in lower fertilization rates (2.04% and 16.90% respectively). No significant differences were found among four fresh sperm doses compared. Mean hatch and survival rates resulting from fresh and cryopreserved sperm were similar. The experiment shows that while only 1.89 × 10⁶ fresh spermatozoa was required to fertilize a fresh egg, 6.94 × 10⁶ (or 3.67 times more) cryopreserved sperm was required to achieve the same level of fertilization. This provides important information for making decision to cryopreserve sperm for commercial and/or conservation purposes.     

Vaccination of Mixed and Full-Sib Families of Channel Catfish Ictalurus punctatus Against Enteric Septicemia of Catfish With a Live Attenuated Edwardsiella ictaluri Isolate (RE-33)

These studies were conducted to evaluate the efficacy of a live attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish. In one study channel catfish fingerlings (72 d of age post hatch) were immersed for 30 min in water containing E. ictaluri RE-33 at dosages of 1 × 10⁶, 1 × 10⁷ and 2 × 10⁷ CFU/ML of water. No mortalities were observed following vaccination. Following exposure to virulent Edwardsiella ictaluri the cumulative mortality of fish vaccinated with dosages of at least 1 × 10⁷ CFU/mL were significantly lower than that of non-vacccinated fish in both laboratory and field challenges. Vaccination with 1 × 10⁶ CFU RE-33mL provided some protection during the laboratory challenge but failed to protect fish under field conditions. In a second study, vaccination of 6 full-sib families of channel catfish at a vaccine dosage of 1 × 10⁷ CFU/mL resulted in a relative percent survival among families ranging from 67.1 to 100%. Significant differences in mortality were found among families and between vaccinated and unvaccinated groups, but there was no family by vaccine interaction. Families with the highest mortality after vaccination were also shown to have the highest mortality without vaccination (r = 0.82; P = 0.04).     

Requirement of n-3 long chain polyunsaturated fatty acids for European sea bass (Dicentrarchus labrax) juveniles: growth and fatty acid composition

European sea bass juveniles (14.4±0.1 g mean weight) were fed diets containing different levels of fish oil then of n-3 highly unsaturated fatty acids (n-3 HUFA) for 12 weeks. The fish performance as well as fatty acid (FA) composition of neutral and polar lipids from whole body after 7 and 12 weeks feeding were studied. The requirements of juvenile sea bass for n-3 highly unsaturated fatty acids (n-3 HUFA) were studied by feeding fish diets containing six different levels of n-3 HUFA ranging from 0.2% to 1.9% of the diet, with approximately the same DHA/EPA ratio (1.5:1).

The growth rate at the end of the trial showed significant differences. Fish fed low dietary n-3 HUFA (0.2% DM of the diet) showed significantly lower growth than the diet 3 (0.7%), then no further improvement (P>0.05) of growth performance was seen by elevating the n-3 HUFA level in the diet up to 1.9% (diet 6). No difference in feed efficiency, protein efficiency ratio or protein retention was observed among treatments, nor in protein and total lipid content. However, the n-3 HUFA levels in diets highly influenced fish fatty acid composition in neutral lipid, while polar lipid composition was less affected. Comparison of polar lipid content after 7 or 12 weeks indicated that DHA remained stable at the requirement level, while arachidonic acid decreased with time. Results of this experiment suggest that the requirement for growth of n-3 HUFA of juvenile sea bass of 14 g weight is at least 0.7% of the dry diet.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.     

Evaluation of carbohydrate rich diets through common carp culture in manured tanks

Four diets (T₀–T₃) were formulated reducing the fishmeal (Indian) component by 100 g kg^–1 from 300 to 0 g kg^–1 and including proportionately increasing quantities of maize. Diets were fed for 120 days at 50 g kg^–1 body weight to triplicate groups of common carp (av. wt. 2.11–2.18 g) stocked at 1 m^–2 in mud bottomed cement tanks (18 m²), fertilized with poultry manure. Fish growth, SGR and FCR in the different treatments were statistically not significantly different ( P  > 0.05). PER was lowest for the 300 g fishmeal kg^–1 diet treatment (diet T₀), increasing with decrease in dietary fishmeal content (diets T₁–T₃). Fish survival ranged from 96.29 to 100%. Diets influenced carcass composition and digestive enzyme activity. A significant increase in lipid deposition was recorded with increasing dietary carbohydrate content. Amylase, protease and lipase activities were higher in fish fed with diets T₂ and T₃. The protein sparing effect of dietary carbohydrate and the economic implication of eliminating fishmeal from the diet are discussed.     

Influence of photoperiod, frequency of stripping and presence of females on sperm output in turbot, Scophthalmus maximus (L.)

Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 10⁹ spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 10⁹ spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 10⁹ spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 10⁹ spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).     

Changes in skin colour and cortisol response of Australian snapper Pagrus auratus (Bloch & Schneider, 1801) to different background colours

A two-factor experiment was carried out to investigate the change in skin colour and plasma cortisol response of cultured Australian snapper Pagrus auratus to a change in background colour. Snapper (mean weight=437 g) were held in black or white tanks and fed diets containing 39 mg unesterified astaxanthin kg⁻¹ for 49 days before being transferred from white tanks to black cages (WB) or black tanks to white cages (BW). Skin colour values [ L ^* (lightness), a ^* (redness) and b ^* (yellowness)] of all snapper were measured at stocking ( t =0 days) and from cages of fish randomly assigned to each sampling time at 0.25, 0.5, 1, 2, 3, 5 and 7 days. Plasma cortisol was measured in anaesthetized snapper following colour measurements at 0, 1 and 7 days. Fish from additional black-to-black (BB) and white-to-white (WW) control treatments were also sampled for colour and cortisol at those times. Rapid changes occurred in skin lightness ( L ^* values) after altering background colour with maximum change in L ^* values for BW and WB treatments occurring within 1 day. Skin redness ( a ^*) of BW snapper continued to steadily decrease over the 7 days ( a ^*=7.93 × e^{−0.051 × time}). Plasma cortisol concentrations were highest at stocking when fish were held at greater densities and were not affected by cage colour. The results of this study suggest that transferring dark coloured snapper to white cages for 1 day is sufficient to affect the greatest benefit in terms of producing light coloured fish while minimizing the reduction in favourable red skin colouration.     

Effects of dietary protein and lipid level on growth and body composition of juvenile ayu (Plecoglossus altivelis) reared in seawater

A 3 (protein levels, 380, 460 and 520 g kg^–1 diet) × 2 (lipid levels, 65 and 140 g kg^–1 diet) factorial experiment with three replicates was conducted. Weight gain, feed efficiency and daily feed intake were not significantly affected by dietary protein level, but were by dietary lipid level. Weight gains of fish fed 65 g lipid kg^–1 diet were significantly, or slightly, higher than for 140 g lipid kg^–1 diet at all protein levels. Daily protein intake was significantly affected by both dietary protein and lipid levels ( P  < 0.002). Daily lipid intake was not significantly affected by dietary protein level, but was by dietary lipid level ( P  < 0.001). Protein efficiency ratio was significantly affected by dietary protein level ( P  < 0.02), but not by dietary lipid level. Protein efficiency ratio tended to improve with the decrease of dietary protein level at the same lipid level. Moisture, protein and lipid contents of whole fish were significantly affected by dietary lipid level ( P  < 0.01). Increased dietary lipid did not improve growth or feed efficiency, but increased body fat deposition. It was concluded that the optimum dietary protein and lipid level for growth of juvenile ayu may be 380 and 65 g kg^–1 diet, respectively, when fish were fed to satiety three times daily in seawater.     

Assessment of cholesterol requirements in the prawn, Penaeus japonicus

Two feeding experiments were conducted to assess the quantitative cholesterol requirement of the juvenile prawn, Penaeus japonicus . The prawns were fed casein-based (experiment 1) and casein- or crab protein-based (experiment 2) diets with or without supplemental cholesterol. The daily increases in quantities of body cholesterol (mg kg⁻¹ prawn day⁻¹) at maximum growth and dietary cholesterol intake over a 40-day feeding period were determined. Prawns fed 5 g kg⁻¹ supplemental cholesterol, regardless of protein sources, gave the highest weight gain. Body retention efficiencies of dietary cholesterol (cholesterol retained × 100/cholesterol intake) vary among prawns fed 5 g kg⁻¹ supplemental cholesterol (diets 3, 5 and 7) but dietary cholesterol requirements expressed as mg kg⁻¹ body weight (BW) day⁻¹ were not significantly different (180 to 200 mg kg⁻¹ BW day⁻¹). Based on dietary cholesterol requirement, the optimum dietary cholesterol levels for the juvenile prawns were estimated in relation to feeding levels. When feeding levels were 3%, 5% and 7% of body weight, optimum dietary cholesterol levels were 5.0 to 6.0, 3.6 to 4.0, and 2.6 to 2.9 g kg⁻¹ of dry diet, respectively. The present study showed the advantages of determining daily cholesterol requirement (mg kg⁻¹ BW day⁻¹) at maximum growth through a factorial method in determining optimum dietary cholesterol levels in P. japonicus .     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.