Effect of soybean meal,raffinose and stachyose on the growth,body composition,intestinal morphology and intestinal microflora of juvenile allogynogenetic silver crucian carp (Carassius auratus gibelio♀×Cyprinus carpio♂)

The study was conducted to investigate the effects of soybean meal (SBM), raffinose and stachyose on juvenile crucian carp (Carassius auratus gibelio♀ × Cyprinus carpio♂). The experimental diets consisted of one control diet based on fish meal (FM), one diet containing 300 g kg^?1 SBM and four FM‐based diets with the addition of either 6.7 g kg^?1 raffinose (Raf), 33.9 g kg^?1 stachyose (Sta), a combination of raffinose and stachyose (Raf?Sta) and finally a Raf?Sta diet supplemented with 2.5 g kg^?1 saponins (Raf?Sta?Sap). After 3 weeks of feeding, the relative gut lengths of SBM‐fed fish and the fish fed stachyose‐containing diets were shorter than those of the FM‐fed fish; further, more SBM‐fed fish showed fissures on the tips of the intestinal folds. After 8 weeks of feeding, the growth of SBM‐fed fish was significantly lower than that of FM‐fed fish (P<0.05). The fish fed Raf?Sta?Sap had a low relative gut length (P<0.05). In comparison with the other fish, the SBM‐fed fish had a higher number of large‐sized homogeneous vacuoles in the cytoplasm of epithelial cells and shorter microvilli. No significant difference was observed in body composition or intestinal microflora. The results indicated that raffinose and stachyose played no or only minor roles in the development of soybean‐induced growth reduction.     

Evaluation of l-ascorbyl-2-glucose as the source of vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf)

This study was conducted to compare l ‐ascorbyl‐2‐glucose (AA2G) with l ‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) for supplying the dietary vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf). Fish were fed one of seven semi‐purified diets containing equivalents of 0, 50, 100 and 200 mg ascorbic acid (AA) kg^?1 diet in the form of AA2G or AMP‐Na/Ca for 12 weeks. After 12 weeks of feeding, weight gain, feed efficiency ratio and survival of fish fed the vitamin C‐free diet were significantly lower than those of fish fed the vitamin C‐supplemented diets in either form. The hepatosomatic index, condition factor and survival of fish fed AMP‐Na/Ca₁₀₀, AMP‐Na/Ca₂₀₀, AA2G₁₀₀ and AA2G₂₀₀ diets were significantly higher than fish fed the vitamin C‐free diet. After 9 weeks of feeding, fish fed the vitamin C‐free diet began to show vitamin C deficiency signs such as anorexia and lethargy. At the end of the 12‐week feeding trial, fish fed the vitamin C‐free diet exhibited vitamin C deficiency signs, e.g., anorexia, scoliosis, exophthalmia and fin haemorrhage. Vitamin C retention in the muscle and liver was similar to those of fish fed AA2G‐ or AMP‐Na/Ca‐supplemented diets. In general, there was no significant difference in the muscle and liver vitamin C concen‐tration in fish fed the AA2G and AMP‐Na/Ca diets at the same supplementation levels.     

Evaluation of dietary yellow loess as an antibiotic replacer on growth,immune responses,serological characteristics and disease resistance in rainbow trout,Oncorhynchus mykiss

A 12 weeks of feeding trial was conducted to evaluate the effects of different levels of dietary yellow loess as an antibiotic (oxytetracycline) replacer in rainbow trout, Oncorhynchus mykiss. Five experimental diets were formulated to contain no antibiotics or yellow loess (control/CON), three graded levels of yellow loess 5 (YL₅), 10 (YL₁₀) and 20 g YL kg^?1 diet (YL₂₀) and oxytetracycline at 5 g OTC kg^?1 diet. Forty‐five fish averaging 39.4 ± 1.6 g (mean ± SD) were randomly distributed in to 15 aquaria. Triplicate groups of fish were fed one of the experimental diets at 1.5 ~ 1.9% of wet body weight per day. At the end of the feeding trial, average weight gain (WG) and specific growth rate (SGR) from fish fed CON diet were significantly lower than those from fish fed YL₁₀, YL₂₀ or OTC diets (P < 0.05). Lysozyme activity from fish fed YL₂₀ was detected to be significantly higher than that from fish fed CON diet (P < 0.05). While, superoxide dismutase (SOD) activity from fish fed YL₁₀ and YL₂₀ was recorded to be significantly higher than that from fish fed CON diet (P < 0.05). Fourteen days of challenge test with bacteria A. salmonicida showed significantly lower survival rate for CON than those of fish fed other experimental diets. Therefore, these results indicated that dietary yellow loess at 10–20 g kg^?1 could be a promising alternative of oxytetracycline in rainbow trout.     

Nitrogen and phosphorus digestibility and excretion of different-sized groups of milkfish (Chanos chanos Forsskal) fed formulated and natural food-based diets

This study determined the digestibility of nitrogen and phosphorus, and the excretion rate of different‐sized groups of milkfish fed a commercial diet, a SEAFDEC formulated diet or lab‐lab (natural food‐based diet). Fish (31.2–263.0 g) were stocked in 12 units of 300‐L fibreglass tanks filled with aerated seawater. The postprandial total ammonia‐nitrogen (TAN) and phosphate (PO₄‐P) excretion of fish were estimated from changes in TAN and PO₄‐P concentrations in water for 24 h. Digestibility was determined from the nitrogen, phosphorus and Cr₂O₃ content of the diets, and pooled faeces after the fish had been fed diets marked with chromic oxide. TAN excretion rate (mg TAN kg^?1 fish day^?1) was significantly lowest (P < 0.05) in medium to very big fish fed the lab‐lab diet (60.8–124.4) and highest in small and medium fish fed the SEAFDEC diet (333.3–331.6) and small fish fed the commercial diet (280.1). Regardless of size, fish fed lab‐lab excreted (mg PO₄‐P kg^?1 fish day^?1) significantly lower PO₄‐P (36.2) but did not differ with fish fed the commercial diet (64.8). Excretion rates decreased exponentially as fish weight increased but positively increased with feed ration. Excretion pattern of milkfish revealed two peaks: the first peak occurred 6 h after feeding and the second peak at 18 h for TAN and 21 h for PO₄‐P, coinciding with the start of the daylight hours. TAN and PO₄‐P excretion accounted for 20.5–34.6% of total N consumed and 18.7–42.6% of P consumed respectively. Approximately 27.9–42.5% of N consumed and 47.2–58.5% of P consumed were lost as faeces. Total nutrient losses were lower using the lab‐lab diet (0.31 g N and 0.14 g P kg^?1 fish) compared with the formulated diets (0.47–0.48 g N and 0.17–0.19 g P kg^?1 fish); the losses decreased per kg of fish as fish size increased. Results suggest that the diet and size of fish influence wastage of N and P to the environment with greater losses in small fish and when artificial diets are used. Such measurements will provide valuable information for the preparation of N and P budgets for milkfish in grow‐out systems.     

Low levels of Aflatoxin B1 could cause mortalities in juvenile hybrid sturgeon,Acipenser ruthenus ♂×A. baeri♀

Toxicity of aflatoxin B₁ (AFB₁) was investigated in juvenile hybrid sturgeon Acipenser ruthenus ♂ × A. baeri♀, an important coldwater finfish farmed in China and other countries. Seven experimental diets (Diet A–G) containing different levels of AFB₁ (0, 1, 5, 10, 20, 40 and 80 μg kg^?1 diet) were fed to juvenile sturgeon weighing 10.53 ± 0.17 g kg^?1 to determine its effect on survival, growth, feed consumption, hematocrit, liver histology as well as muscular and hepatic toxin accumulation. The experiment lasted for 35 days and was conducted in two periods of 25 and 10 days each. No external changes or unusual behaviour was observed in the fish fed diets with AFB₁. Mortality was observed in fish fed with highest levels of AFB₁ (80 μg kg^?1– Diet G) from day 12 onwards. After 25 days, fish fed the diet of 80 μg AFB₁ kg^?1 showed significant lower survival (50 ± 5.77%) followed by those fed 40 μg AFB₁ kg^?1 diet (80 ± 5.77%) and 20 μg AFB₁ kg^?1 diet (86.66 ± 3.33%). No significant difference was observed in specific growth rate (SGR) or hepatosomatic index (HSI) between groups. Hematocrit was significantly higher in the fish fed the diet of highest AFB₁. The fish were weighed at day 25 in some treatments (Diets F and G) because of high mortality. However, feeding was continued for another 10 days to observe mortality or behavioural changes if any in the other groups. After 35 days, survival in the fish fed Diet F (40 μg AFB₁ kg^?1) was 40% and those fed Diet E (20 μg AFB₁ kg^?1) was 36.2%. Significant histopathological changes including nuclear hypertrophy, hyperchromasia, extensive biliary hyperplasia, focal hepatocyte necrosis and presence of inflammatory cells were observed in the liver of fish fed high levels of aflatoxin (40 and 80 μg kg^?1). AFB₁ accumulation in fish muscle and liver increased with increased dietary AFB₁ levels. It could be confirmed that 10 μg AFB₁ kg^?1 diet was the maximum allowable level in hybrid sturgeon diet.     

Effects of different dietary levels of L-ascorbyl-2-polyphosphate on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel)

This experiment was conducted to study the effects of different dietary levels of vitamin C, L‐ascorbyl‐2‐polyphosphate (ASPP), on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Fish were fed one of six semi‐purified diets containing an equivalent of 0, 25, 50, 75, 150, or 1500 mg ascorbic acid (AA) kg^?1 diet (C₀, C₂₅, C₅₀, C₇₅, C₁₅₀ or C₁₅₀₀) in the form of ASPP for 12 weeks. Weight gain (WG) and protein efficiency ratio (PER) of fish fed the C₀ diet were significantly lower than those of fish fed the other diets (P < 0.05), and WG and PER of fish fed the C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀₀ diet (P < 0.05). Fish fed the C₀ diet exhibited vitamin C deficiency symptoms such as anorexia, scoliosis, cataract, exophthalmia and fin hemorrhage at the end of the 12‐week test. After 12 weeks of the feeding trial, AA concentrations from gill, kidney, and liver of fish fed the C₀, C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀ and C₁₅₀₀ diets (P < 0.05). Based on broken line analyses for WG and PER, the optimum dietary levels of vitamin C were 91 and 93 mg AA kg^?1 diet respectively. These findings suggest that the dietary vitamin C requirement could be 93 mg AA kg^?1 diet to support reasonable growth, and greater than 150 mg AA kg^?1 diet may be required for AA saturation of major tissues for juvenile olive flounder under experimental conditions.     

Effects of canola meal on physiological and biochemical parameters in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (initial body weight 4.16 ± 0.25 g) were fed diets [crude protein 420 g kg^?1; gross energy 18.7 MJ kg^?1 dry matter (DM); crude fat 110 g kg^?1] containing graded levels of either a canola meal (crude protein 350 g kg^?1 DM) supplemented with DL‐methionine as partial fish meal protein. A growth trial was conducted over 16 weeks at a water temperature of 12 ± 1 °C. At the end of the growth trial, in addition to body composition analyses, plasma tri‐iodothyronine (T₃) and thyroxine (T₄), cholesterol and liver fatty acid composition were measured. Replacement of fish meal with canola meal (100–570 g kg^?1 replacement) did not affect on growth performance. At 16th week, plasma cholesterol levels were reduced in fish fed all diets in comparison with 8th week. Plasma T₄ levels were significantly higher in the canola meal‐fed fish sampled after 16 weeks, but no significant differences in T₃ levels were obtained (P > 0.05). Proximate compositions were affected by dietary treatments. The liver fatty acid composition reflected that of the diet with a higher level of polyunsaturated (n‐6) fatty acids in fish fed diet canola meal and a higher content in n‐3/n‐6 ratio in fish fed diet without canola meal. These studies show that canola meal has potential to replace substantial levels of fish meal in diets for carnivorous fish without compromising performance.     

Use of probiotics to enhance growth,stimulate immunity and confer disease resistance to Aeromonas salmonicida in rainbow trout (Oncorhynchus mykiss)

An 8‐week feeding trial was conducted to evaluate the effects of dietary probiotics on growth, non‐specific immune responses and disease resistance in juvenile rainbow trout, Oncorhynchus mykiss. Fish averaging 5.8 ± 0.8 g (mean ± SD) were fed one of the five experimental diets; one control (Cont), and four other diets were prepared by supplementing single probiotics 1 (Bacillus subtilis; SP₁, 0.5%), single probiotics 2 (Bacillus licheniformis; SP₂, 0.5%), multi‐probiotics (B. subtilis + B. licheniformis; MP, 0.5%) and oxytetracycline (OTC) at 5 g OTC kg^?1 diet. After 8 weeks of the feeding trial, weight gain and specific growth rate of fish fed SP₁, SP₂ and OTC diets were significantly higher than those of fish fed Cont diet (P < 0.05). Superoxide dismutase (SOD) and lysozyme activities of fish fed SP₁, SP₂ and MP diets were significantly higher than those of fish fed Cont diet (P < 0.05). There were no significant differences in SOD and lysozyme activities among fish fed SP₁, SP₂, MP and OTC diets. In challenge test with Aeromonas salmonicida for 15 days, fish fed SP₁, SP₂ and MP diets showed significantly higher cumulative survival rate than those of fish fed Cont diet (P < 0.05). However, there were no significant differences in cumulative survival rate among fish fed SP₁, SP₂, MP and OTC diets. Although there was a little advantage in fish fed MP diet in terms of non‐specific immune responses, single or multi‐probiotics are equally effective statistically. These results indicate that single or multi‐probiotics had equal beneficial effects as an antibiotic replacer on growth performance, non‐specific immune responses and disease resistance in juvenile rainbow trout.     

Effect of feeding level on growth and nutrient deposition in rainbow trout (Oncorhynchus mykiss Walbaum) growing from 150 to 600 g

The effects of feeding level on growth and energy partitioning were studied in rainbow trout growing from 150 to 600 g. Triplicate groups of fish (initial weight 158 g fish^?1) were fed a practical diet at various feeding levels (25%, 50%, 75% and 100% of near satiation) for 24 weeks at 8.5°C. The final body weights of fish were 235, 381, 526 and 621 g. Restricted feeding levels significantly reduced live weight gain. Feeding levels had less pronounced effects on feed efficiency ratio, which were 0.98, 1.08, 1.02 and 0.83, respectively, for the 25%, 50%, 75% and 100% feeding levels. The growth of fish fed to near satiation was accurately described by the thermal‐unit growth coefficient. The growth data also showed that the widely used specific growth rate was not an appropriate model. Fish fed at the lowest feeding level (25%), which represented a maintenance ration (energy gain was less than 2 kJ fish^?1 day^?1), showed positive protein deposition but negative lipid deposition. This indicates that fish fed a maintenance ration mobilize body lipid reserve to support protein deposition. The efficiency of energy for growth (k_g) was estimated to be 0.63. The factorial multiple regression approach estimated that the partial efficiencies of metabolizable energy utilization for protein deposition (k_p) and lipid deposition (k_f) were 0.63 and 0.72, and that maintenance energy requirement was about 19 kJ (kg BW^0.824)^?1, for rainbow trout held at 8.5°C.     

Live Spirulina (Arthrospira platensis) as a growth and immunity promoter for Nile tilapia, Oreochromis niloticus (L.), challenged with pathogenic Aeromonas hydrophila

The use of antibiotics to control fish disease is one of the constraints in tilapia farming. Hence, the use of probiotic and live organisms as alternative strategies has received much more attention. This study was undertaken to evaluate the use of Spirulina (Arthrospir platensis) as a growth and immunity promoter for Nile tilapia, Oreochromis niloticus (L.). Fish (1.88 g) were randomly distributed at 20 fish per 100 L aquarium and fed a diet containing either 0.0, 1.25, 2.5, 5.0, 7.5 or 10.0 g Spirulina kg^?1 diet for 12 weeks. After the feeding trial, fish of each treatment were challenged by pathogenic Aeromonas hydrophila, which was given by an interperitoneal injection, and they were kept under observation for 10 days to record any abnormal clinical signs and the daily mortality rate. The growth‐promoting influence of Spirulina was observed with fish, and the optimum growth and feed utilization were obtained at 5.0 g Spirulina kg^?1 diet. No significant changes were observed in fish survival among the different treatments. Spirulina supplementation improved the protein and lipid contents in fish bodies without significant differences among them, especially when fed 2.5–10.0 g kg^?1 diet, but no significant differences in the moisture and ash contents were observed among different treatments. The physiological parameters were improved when fish were fed a Spirulina‐enriched diet. However, the highest red blood cells, white blood cells and nitro blue tetrazolium values were obtained at 5.0–10.0 g Spirulina kg^?1 diet; meanwhile, the lowest value was obtained in the control. Total fish mortality due to A. hydrophila infection decreased with an increase in the Spirulina level in fish diets. These results indicate that Spirulina supplement is promising as an alternative method to antibiotics for disease prevention in tilapia aquaculture, and the optimum level of Spirulina in the fish diet should be 5.0–10.0 g kg^?1 diet.     

Dietary levamisole modulates the immune response and disease resistance of Asian catfish Clarias batrachus (Linnaeus)

In order to determine the immunomodulatory effect of dietary levamisole in Asian catfish (Clarias batrachus), fish were fed four different diets for 10 days: a formulated diet as control and the same diet supplemented with 50, 150 or 450 mg levamisole kg^?1 feed. The serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity, serum haemagglutination titre, natural haemolytic complement activity (ACH₅₀), myeloperoxidase and lysozyme activities, total protein level and oxidative radical production by neutrophils as a measure of non‐specific immunity as well as disease resistance against A. hydrophila challenge to separate vaccinated and non‐vaccinated groups were evaluated at 0, 1, 2 and 3 weeks after last administration of levamisole. Levamisole supplement at the lowest level (50 mg kg^?1) significantly enhanced oxidative radical production and serum myeloperoxidase (MPO) content immediately after 10 days of feeding, which reached peak values after 3 and 2 weeks of feeding respectively. Haemolytic complement and haemagglutination titre were significantly enhanced after 3 and 1 weeks respectively. Haemolytic complement activity and MPO activities were significantly raised to 150 mg kg^?1 after 3 and 2 weeks, respectively. At the highest level of levamisole feeding (450 mg kg^?1) significant decreases in superoxide production and complement activity were measured immediately after levamisole feeding, which returned to the normal level after 1 week post‐ feeding. Fish were challenged with a virulent strain of A. hydrophila at 0, 1, 2 and 3 weeks after levamisole feeding, and the cumulative per cent survival was recorded over 10 days. Feeding levamisole at 50, 150 or 450 mg kg^?1 increased per cent survival in vaccinated fish immediately after levamisole feeding, and survival was significantly higher at 450 mg kg^?1. There was no difference in mortality patterns in non‐vaccinated fish. The results support the use of levamisole at 50 mg kg^?1 feed for 10 days as an immunostimulant in Asian catfish farming.     

Separate and combined effects of cyclic fasting and l‐carnitine supplementation in red porgy (Pagrus pagrus,L. 1758)

We examined the effects of cyclic fasting in red porgy (Pagrus pagrus) fed different dietary carnitine levels. Juvenile fish (23.58 ± 3.49 g) were divided into eight groups – four groups were fed every day to apparent satiation, while the other four were fasted for 7 days every 2 weeks. In each feeding regime, two replicates were fed an l ‐carnitine non‐supplemented diet (46 mg kg^?1) and the other two groups were fed an l ‐carnitine supplemented diet (630 mg kg^?1). Fish fed 630 mg l ‐carnitine accumulated two times more l ‐carnitine in muscle than fish fed 46 mg l ‐carnitine. Cyclic fasting reduced the growth performance and lipid content in the liver. Carnitine supplementation did not affect performance and body composition, but decreased the n‐6 PUFA content. Moreover, the combined effects of fasting and carnitine supplementation were observed on reducing the n‐3 fatty acid content. Areas of steatosis were found in the livers of red porgy, but the results revealed that supplementation of l ‐carnitine in cyclic fasted fish contributed towards a lower degree of vacuolization than in fish fed to apparent satiation. Regardless of the feeding regime applied, the spleen of fish fed the l ‐carnitine‐supplemented diet was haemorrhagic and hyper activation of melanomacrophage cells was observed.     

Single or combined effects of fructo‐ and mannan oligosaccharide supplements and Bacillus clausii on the growth,feed utilization,body composition,digestive enzyme activity,innate immune response and lipid metabolism of the Japanese flounder Paralichthys olivaceus

In this study, we examined the effects of the following eight experimental diets, which varied in fructo oligosaccharides (FOS), mannan oligosaccharides (MOS) and Bacillus clausii concentrations, on the Japanese flounder: control diet (no FOS, MOS and B. clausii), diet F (5 g kg⁻¹ FOS), diet M (5 g kg⁻¹ MOS), diet FM (2.5 g kg⁻¹ FOS + 2.5 g kg⁻¹ MOS), diet B (10⁷ cells g⁻¹B. clausii), diet FB (5 g kg⁻¹ FOS + 10⁷ cells g⁻¹B. clausii), diet MB (5 g kg⁻¹ MOS + 10⁷ cells g⁻¹B. clausii) and diet FMB (2.5 g kg⁻¹ FOS + 2.5 g kg⁻¹ MOS + 10⁷ cells g⁻¹B. clausii). Japanese flounder, initially weighing an average of 21 g, were distributed into 24 net cages at a stocking density of 20 fish per cage. Each diet was hand‐fed to three groups of fish twice daily for 56 days. The weight gain rate (WGR) in fish fed diets B, MB and FMB were significantly higher than in fish fed the control diet, where the fish fed diet FMB had the highest WGR. Fish fed any of the diets, except diets F and B, exhibited better feed conversion ratio than those fed the control diet. Diets MB and FMB significantly elevated intestinal protease activity compared with the control diet, but only the diet FMB promoted amylase activity. Feeding diets FB and FMB increased body protein deposition; additionally, feeding diets B, MB and FMB significantly reduced body lipid deposition. Lysozyme (LSZ) activity was significantly higher in fish fed diets B, FB, MB and FMB than in fish fed the control diet. All diets, except diet M, decreased triglyceride (TG) levels compared to the control diet. Low‐density lipoprotein cholesterol levels in fish fed diets F, FB and FMB were significantly lower than in fish fed the control diet. Without exception, no diets affected feeding rate, condition factor, body moisture, ash contents, phagocytic activity of leucocytes or cholesterol or high‐density lipoprotein cholesterol levels. Our results suggest that diets supplemented with FOS, MOS and B. clausii improved growth performance and health benefits of the Japanese flounder more than other diets or the control diet.     

Dietary calcium requirements of juvenile tilapia, Oreochromis niloticus × O. aureus, reared in fresh water

A feeding experiment was conducted to determine the dietary calcium (Ca) requirement for juvenile hybrid tilapia, Oreochromis niloticus × O. aureus reared in nature water. Purified diet supplemented with 0, 1, 2, 3, 4, 5, 7 and 10 g Ca kg⁻¹ diet providing of 0.6, 1.6, 2.6, 3.7, 4.7, 5.5, 7.5 and 10.7 g Ca kg⁻¹ diet, respectively, were fed to tilapia (mean initial weight: 0.52 ± 0.01 g, n = 3) for 8 weeks. Each diet was fed to three replicate groups of fish in a closed, recirculating fresh water rearing system. The rearing water contained 27.1–33.3 mg L⁻¹ Ca. The tilapia fed the diets supplemented with ≥3.7 g Ca kg⁻¹ had significantly (P < 0.05) higher weight gain, when compared with fish fed the diet with ≤1.6 g Ca kg⁻¹. Fish fed the unsupplemented control showed significantly lower weight gain when compared with the other groups (P < 0.05). Bone Ca concentration was highest in fish fed the diets with ≥4.7 g Ca kg⁻¹, intermediate in fish fed the diet with 2.6 g Ca kg⁻¹ and lowest in fish fed the control diet. Scale Ca concentration was higher in fish fed the diets with ≥3.7 g Ca kg⁻¹ than in fish fed the diets with ≤2.6 g Ca kg⁻¹. Serum alkaline phosphatase activity was 36% increased in fish fed the diets with ≥2.6 g Ca kg⁻¹ than fish fed the diets with <1.6 g Ca kg⁻¹. Analysis by broken‐line regression of weight gain, bone and scale Ca concentrations indicated that the adequate dietary Ca concentration for tilapia in water containing 27.1–33.3 mg Ca L⁻¹ was 3.5, 4.3 and 4.2 g Ca kg⁻¹ diet, respectively, supplied as Ca‐lactate.     

Evaluation of optimum dietary protein-to-energy ratio in juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel)

An 8‐week feeding trial was conducted to estimate the optimum dietary protein to energy (P/E) ratio in juvenile olive flounder Paralichthys olivaceus. Eight experimental diets were formulated with two energy levels and four protein levels at each energy level. Two energy levels of 12.5 and 16.7 kJ g^?1 diets were included at crude protein (CP) levels of 25%, 30%, 35% and 45% with 12.5 kJ g^?1, and CP levels of 35%, 45%, 50% and 60% with 16.7 kJ g^?1. After 1 week of the conditioning period, fish initially averaging 8.1±0.08 g (mean±SD) were randomly distributed into the aquarium as groups of 15 fish. Each diet was fed on a dry‐matter basis to fish in three randomly selected aquariums at a rate of 3–5% of total wet body weight per day for 8 weeks. After 8 weeks of the feeding trial, weight gain (WG), feed efficiency ratio and specific growth rate of fish fed 45% CP with 16.7 kJ g^?1 energy diet were significantly higher than those from the other dietary treatments (P<0.05). WG of fish fed 12.5 kJ g^?1 energy diets increased with the increase of dietary protein levels. However, WG of fish fed 16.7 kJ g^?1 energy diets increased with the increase of dietary protein levels up to 45% CP and then decreased when fish fed 50% and 60% CP diets. Both dietary protein and energy affected protein retention efficiency and energy retention efficiency. Haemoglobin (Hb) of fish fed 35% and 45% CP diets with 12.5 kJ g^?1 energy were significantly high and not different from Hb of fish fed 45% and 50% CP diets with 16.7 kJ g^?1 energy. Haematocrit of fish fed 45% CP diet with 16.7 kJ g^?1 energy was significantly higher than those from fish fed 25% and 30% CP diets with 12.5 kJ g^?1 energy (P< 0.05). Based on the results of this experiment, we concluded that the optimum dietary P/E ratio was 27.5 mg protein kJ^?1 with diet containing 45% CP and 16.7 kJ g^?1 energy in juvenile olive flounder.     

Dietary copper effects survival of channel catfish challenged with Flavobacterium columnare

Columnaris disease is an important bacterial disease of commercially grown channel catfish, Ictalurus punctatus. Copper sulphate (CuSO₄) has been shown to be therapeutic and prophylactic as a water treatment for columnaris disease. Copper is an essential dietary component in animal feeds and CuSO₄ is typically included in base diets; a study was conducted to evaluate whether fish feed supplemented with additional CuSO₄ at 0, 40 and 80 mg kg^?1 of diet and fed at a daily rate of 3% body weight would affect survival to columnaris disease. Results indicate fish fed the copper‐supplemented diet for 2 weeks significantly increased survival following F. columnare challenge. This increase appeared to be dose‐dependent. The mean per cent survival (±SEM) for fish fed the base diet (unsupplemented) for 2 weeks and then challenged was 2.0% ± 1.1. Fish fed the base plus 40 mg CuSO₄ kg^?1 had a mean survival of 22.0% ± 11.0. Fish fed the base plus 80 mg CuSO₄ kg^?1 had a mean survival of 29.3% ± 13.4. The mean per cent survival for fish fed the base diet for 4 weeks and then challenged was 28.3% ± 9.0. Fish fed the base plus 40 mg CuSO₄ kg^?1 for 4 weeks had a mean survival of 12.5% ± 6.3. Fish fed the base plus 80 mg CuSO₄ kg^?1 for 4 weeks had a mean survival of 40.5% ± 8.1. There was a significant effect after 4 weeks with fish fed the base plus 80 mg CuSO₄ kg^?1 mg not with 40 mg kg^?1.     

Effects of dietary blend of fish oil with corn oil on growth and non-specific immune responses of grouper, Epinephelus malabaricus

A growth trial was conducted to investigate the effects of fish oil and corn oil on the growth and non‐specific immune responses of juvenile grouper, Epinephelus malabaricus. Five semi‐purified diets were supplemented with 40 g kg⁻¹ of either fish oil (F4), corn oil (C4) or blend of fish oil with corn oil at ratio of 3 : 1 (F3C1); 1 : 1 (F2C2) and 1 : 3 (F1C3). Each diet was fed to triplicate groups of grouper (mean initial weight: 10.26 ± 0.14 g) in a recirculating rearing system for 8 weeks. Weight gain and feed efficiency of fish fed the F4, F3C1 and F2C2 diets were the highest (P < 0.05), followed by fish fed the F1C3 diet, and the lowest in fish fed the C4 diet. Fish fed the C4 diet had a lower survival rate than fish on other dietary treatments. Fatty acid composition of liver and muscle in fish generally reflected the composition of the diet. Leukocyte superoxide anion (O⁻₂) production ratio was the highest in fish fed the F3C1 and F2C2 diets, followed by fish fed the F4 and F1C3 diets, and the lowest in fish fed the C4 diet. Fish fed the F3C1 and F2C2 diets had higher plasma lysozyme activities than fish fed the F4 and C4 diets. Plasma alternative complement activity was higher in fish fed the F3C1, F2C2 and F1C3 diets than fish fed the F4 and C4 diets. These results suggest that grouper fed diets with 3 : 1 or 1 : 1 of fish oil to corn oil ratio had similar growth to the fish fed diet with fish oil. Blend of fish oil with corn oil in diet significantly enhanced non‐specific immune responses of grouper when the fed diet contained fish oil as the only lipid source.     

Effects of dietary supplementation with probiotic live yeast Debaryomyces hansenii on the immune and antioxidant systems of leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila

Two trials were conducted to determine the efficacy of fish fed live yeast Debaryomyces hansenii strain CBS 8339 on immune and antioxidant systems in leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila. Juveniles (12±0.5 g) were fed with a control diet or a D. hansenii‐supplemented diet (10⁶ colony‐forming units per gram) for 5 weeks. The live weight of fish was registered on a weekly basis. After 4 weeks, fish from each treatment were immunocompromised with pathogenic A. hydrophila and further fed for 1 week in order to evaluate the effect on immunological and antioxidant parameters. Generally, the results showed enhanced growth performance in fish fed the diet containing yeast compared with the control. Addition of live yeast had no significant effect on the immunological parameters after 4 weeks of feeding. However, post infection with A. hydrophila fish fed the yeast‐supplemented diet resulted in a significant increase in the levels of plasmatic immunoglobulin M. Superoxide dismutase and catalase (CAT) activities were significantly higher in the yeast group. In this fish, CAT and heat shock protein 70 genes were up‐regulated before and after infection of A. hydrophila. The present study is the first one reporting that yeast (D. hansenii) can enhance immunity and resistance against A. hydrophila.     

Dietary biotin requirement for growth of juvenile zebrafish Danio rerio (Hamilton‐Buchanan)

This study was conducted to estimate the dietary biotin requirement for maximum growth of zebrafish Danio rerio. Six isonitrogenous and isocaloric purified diets containing 0.031 (biotin‐unsupplemented diet), 0.061, 0.263, 0.514, 1.741 and 2.640 mg biotin kg^?1 diet were fed in triplicate tanks for a total of 12 weeks to juvenile zebrafish (initial mean body mass 0.13 ± 0.001 g). From 4 weeks of feeding, fish fed diets with ≤0.061 mg biotin kg^?1 showed biotin deficiency signs, such as retarded growth and skeletal deformity, was observed on some dead fish. At the end of the study, gill disorders were observed on some fish and liver glycogen accumulation were also observed in fish fed these diets. Fish fed the biotin‐unsupplemented diet exhibited a lower final body weight, protein efficiency ratio and feed utilization than the fish fed the biotin‐supplemented diets, whereas the highest values were observed with the diet containing 0.51 mg biotin kg^?1 diet (P < 0.05). A linear relationship (r² = 0.77; P < 0.0001) was observed between whole‐body biotin content and dietary biotin level. A broken‐line analysis indicated that the optimum dietary biotin content for maximal growth expressed as final body weight is 0.51 mg kg^?1 diet.     

Effects of low and medium levels of red alga Nori (Porphyra yezoensis Ueda) in the diets on growth,feed utilization and metabolism in intensively fed Nile tilapia,Oreochromis niloticus (L.)

A 10‐week feeding experiment was conducted in a respirometric system to evaluate the suitability of red alga Nori (Porphyra yezoensis) as a partial replacement for fish meal in intensively fed Nile tilapia. Diets were formulated to be iso‐nitrogenous and iso‐energetic and to replace 0%, 15% and 30% fish meal with Nori, P. yezoensis. No statistically significant differences in growth or feed utilization between the control diet and either Nori diet were found. Tilapia fed on Nori 15 increased their body mass by 415%, those fed the control diet by 341% and fish fed Nori 30 by 317% over the experimental period. Nori 15 fed fish showed the best feed utilization, highest metabolic growth rate and significantly lower oxygen consumption rates than the control. Both Nori supplemented groups (Nori 15: 4.1 g O₂ g^?1 protein gain and Nori 30: 3.9 g O₂ g^?1 protein gain) used less oxygen per gram of protein gain compared to fish fed the control diet (5.4 g O₂ g^?1 protein gain). Our results show that a low replacement level (15%) of fish meal with Nori improves growth and feed utilization while oxygen consumption and amount of oxygen consumed per gram protein gain are significantly decreased.