Modulation of neuronal activity and hepatic metabolism by ploidy and l‐carnitine supplement in rainbow trout (Oncorhynchus mykiss)

l ‐carnitine, a multiphysiological, bioactive and pollution‐free additive, is known to act as a growth‐enhancer and appears to act as an antioxidant compound. However, high dosages may cause detrimental physiological effects to fish. Considering such, a feeding trial was carried out to evaluate the effects of dietary l ‐carnitine content on neuronal activity (acetylcholinesterasic activity), hepatic metabolism (catalase activity) and liver histological markers in both diploid and triploid trout (Oncorhynchus mykiss) juveniles. Fish were hand‐fed for 56 days on three nutritionally identical diets, varying in l ‐carnitine content; unsupplemented diet (15 mg kg^?1) and supplemented (200 or 530 mg kg^?1) diets. Moderate to severe glycogen/lipid depletion was the major liver histopathological disorder observed in all dietary groups. This cause–effect relationship appeared to be dose and time‐dependent, suggesting an important role of l ‐carnitine in the liver metabolism. Moreover, the hepatocite size and nuclear diameter appeared to be larger in l ‐carnitine supplemented groups than in control group. l ‐carnitine caused significant induction of catalase activity and inhibition of AChE. Diploid and triploid trouts showed marked differences in enzymatic activities, reflected by consistently higher values of AChE activities for triploid animals.     

Effects of supplemental dietary l‐carnitine and ractopamine on the performance of juvenile rainbow trout,Oncorhynchus mykiss

This study was carried out to investigate a possible protein‐sparing action of l ‐carnitine and ractopamine in rainbow trout, Oncorhynchus mykiss. An 8‐week feeding trial was carried out to evaluate the effects of supplementation of three levels of l ‐carnitine (0, 1 and 2 g kg^?1) and two levels of ractopamine (0 and 10 mg kg^?1) on growth performance, fillet fatty acid compositions and blood biochemical parameters in a 3 × 2 factorial experimental design. Ractopamine and 1 g kg^?1 carnitine improved the specific growth rate (1.03% and 1.05% day^?1), feed conversion ratio (FCR, 1.3 and 1.29), protein efficiency ratio (PER, 1.88 and 1.85) of fish and crude protein (73.5 and 73.8) content of fish fillet. l ‐carnitine and ractopamine increased the levels of albumin, total protein and globulin in the serum of fish. Apart from eicosapentaenoic acid and docosahexaenoic acid, other fatty acids of fish fillet were increased by ractopamine, while total saturated fatty acids were almost intact. However, the total n‐3 poly unsaturated fatty acids were reduced by l ‐carnitine supplementation (P<0.05). The present study showed that 1 g kg^?1l ‐carnitine and 10 mg kg^?1 ractopamine each can improve the performance of rainbow trout and their combination in diet could enhance the protein level and change the fatty acids profile in fillet muscle.     

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.     

Effect of dietary supplemental l‐carnitine on growth performance,body composition and antioxidant status in juvenile black sea bream,Sparus macrocephalus

Effects of dietary l ‐carnitine were studied in juvenile black sea bream (Sparus macrocephalus). The semipurified basal diet [crude protein 450 g kg^?1 dry matter (DM); crude lipid 126 g kg^?1 DM] was formulated to choose white fishmeal as the protein source and fish oil plus corn oil (1 : 1) as the lipid source. Six diets (control + diets 1–5) containing 0.1, 0.12, 0.16, 0.24, 0.39 and 1.1 g of l ‐carnitine kg^?1 diet were fed to triplicate groups of black sea bream (initial weight 13.10 ± 0.05 g) for 8 weeks. At the end of the feeding trial, growth performance, body composition and antioxidant status were determined. The results showed that relative growth rate (RGR) was significantly improved by the elevation of dietary l ‐carnitine level from 0.1 to 0.24 g kg^?1, but decreased with further increment (P < 0.05). Lipid content decreased significantly (P < 0.05) in the dorsal muscle whereas increased (P < 0.05) in the liver with the addition of dietary l ‐carnitine. Dietary l ‐carnitine supplements elevated enzymatic antioxidants (superoxide dismutase, SOD; catalase, CAT; glutathione‐S‐transferase, GST) activities (P < 0.05) yet decreased the content of non‐enzymatic factor, total sulphydryl groups (TSH) (P < 0.05). In summary, the optimum dietary l ‐carnitine level was 0.284 g kg^?1 diet by second‐polynomial regression analysis based on RGR (y = ?647.4x² +367.97x + 234.55; R² = 0.977, x = dietary l ‐carnitine levels, y = RGR), and dietary l ‐carnitine addition within the levels adopted in our study could depress lipid peroxidation in tissues of juvenile black sea bream.     

Increased dietary phosphorous prevents vertebral deformities in triploid Atlantic salmon (Salmo salar L.)

Triplicate groups of triploid and diploid Atlantic salmon were fed diets with a low (LP, total P: 7.1 g kg^?1), medium (MP, total P: 9.4 g kg^?1) or high (HP, total P: 16.3 g kg^?1) phosphorous (P) level from first feeding (0.18 g) to transfer to sea water (~50 g, duration: 203 days) and subsequently fed a commercial diet in sea water for 426 days (~3 kg). This study examined the short‐ and long‐term effects of dietary P on freshwater performance (mortality, growth), vertebral deformities (radiology), bone cell activity (ALP and TRACP enzyme activity in vertebrae and scales, and fgf23, bgp and igf‐I relative gene expression in vertebrae), bone mineralization (ash content) and some parameters related to fish condition (heart and liver size). Irrespective of ploidy, at seawater transfer, fish fed the MP diet had significantly highest length and weight and those fed the LP diet significantly lowest length and weight, while those fed the HP diet had intermediate lengths and weights. Increased dietary phosphorus reduced deformities in both ploidies at seawater transfer; however, triploids fed the LP and MP diets had more deformities than diploids fed the respective diets, while there was no ploidy effect observed for fish fed the HP diet. The vertebral bone ash content at seawater transfer was significantly higher in diploids than in triploids when fed the MP diet only. Alkaline phosphatase (ALP) and tartrate‐resistant acid phosphatase (TRACP) enzyme activities and relative gene expression of bone hormones involved in metabolism of plasma phosphate (fgf23) and bone growth (bgp) were not affected by ploidy at seawater transfer, but by dietary P level; LP increased ALP activity and reduced TRACP activity and fgf23 and bgp expression levels in vertebral bone. In scales, LP increased both ALP and TRACP activity. At the termination of the seawater period, the group‐wise pattern in occurrence of vertebral deformities was the same as at seawater transfer. The present results on mortality, growth, bone mineralization and development of skeletal deformities all demonstrate that triploids have a higher P requirement than diploids in fresh water. This study shows that an optimalization of P nutrition for triploid Atlantic salmon can improve health and welfare and reduce down‐grading of triploid salmon.     

Dietary tryptophan requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week feeding trial was conducted to evaluate the effects of dietary tryptophan concentration on weight gain and feed efficiencies of fingerling Indian major carp, Cirrhinus mrigala. Six isonitrogenous (40% crude protein) and isocaloric (17.90 kJ g^?1) amino acid test diets containing casein, gelatin and l ‐crystalline amino acids with graded levels of l ‐tryptophan (0.06, 0.16, 0.26, 0.36, 0.46 and 0.56 g 100 g^?1 dry diet) were formulated. Fish (4.25±0.30 cm, 0.62±0.02 g) were randomly stocked in triplicate groups in 70 L (water volume 55 L) flow‐through (1–1.5 L min^?1) indoor circular tanks and fed experimental diets at 5% of their body weight/day in two feedings at 08:00 and 16:00 hours. Maximum live weight gain (277%), lowest feed conversion ratio (FCR) (1.50) and highest protein efficiency ratio (PER) (1.66) were measured at 0.36% dietary tryptophan. The relationship between dietary tryptophan levels and weight gain, FCR and PER data were described using second‐degree polynomial regression analysis indicating the tryptophan requirement at 0.42, 0.39 and 0.38 g 100 g^?1 of dry diet respectively. Whole body moisture decreased with increasing tryptophan up to 0.36%. Significantly (P<0.05) higher protein content was evident in fish fed diet containing 0.36% tryptophan. Body fat increased significantly (P<0.05) in fish fed with different tryptophan concentrations except those fed 0.36% tryptophan where a significantly lower fat content was noted. Significantly (P<0.05) higher ash content was reported at 0.06% and 0.16% tryptophan levels. Survival was 100% in fish fed all the diets except those fed 0.06% tryptophan. Based on the results, diets for fingerling C. mrigala should contain tryptophan at 0.38 g 100 g^?1 dry diet, corresponding to 0.95 g 100 g^?1 dietary protein for optimum growth and efficient feed utilization.     

Influences of dietary fatty acid profile on growth, body composition and blood chemistry in juvenile fat cod (Hexagrammos otakii Jordan et Starks)

This study was conducted to investigate the influence of dietary lipid source and n‐3 highly unsaturated fatty acids (n‐3 HUFA) level on growth, body composition and blood chemistry of juvenile fat cod. Triplicate groups of fish (13.2 ± 0.54 g) were fed the diets containing different n‐3 HUFA levels (0–30 g kg^?1) adjusted by either lauric acid or different proportions of corn oil, linseed oil and squid liver oil at 100 g kg^?1 of total lipid level. Survival was not affected by dietary fatty acids composition. Weight gain, feed efficiency and protein efficiency ratio (PER) of fish fed the diets containing squid liver oil were significantly (P < 0.05) higher than those fed the diets containing lauric acid, corn oil or linseed oil as the sole lipid source. Weight gain, feed efficiency and PER of fish increased with increasing dietary n‐3 HUFA level up to 12–16 g kg^?1, but the values decreased in fish fed the diet containing 30 g kg^?1 n‐3 HUFA. The result of second‐order polynomial regression showed that the maximum weight gain and feed efficiency could be attained at 17 g kg^?1 n‐3 HUFA. Plasma protein, glucose and cholesterol contents were not affected by dietary fatty acids composition. However, plasma triglyceride content in fish fed the diet containing lauric acid as the sole lipid source was significantly (P < 0.05) lower than that of fish fed the other diets. Lipid content of fish fed the diets containing each of lauric acid or corn oil was lower than that of fish fed the diets containing linseed oil or squid liver oil only. Fatty acid composition of polar and neutral lipid fractions in the whole body of fat cod fed the diets containing various levels of n‐3 HUFA were reflected by dietary fatty acids compositions. The contents of n‐3 HUFA in polar and neutral lipids of fish increased with an increase in dietary n‐3 HUFA level. These results indicate that dietary n‐3 HUFA are essential and the diet containing 12–17 g kg^?1 n‐3 HUFA is optimal for growth and efficient feed utilization of juvenile fat cod, however, excessive n‐3 HUFA supplement may impair the growth of fish.     

The effects of dietary vitamin C on growth performance,serum enzymes activities and resistance to Vibrio alginolyticus challenge of yellow drum Nibea albiflora

A 9‐week feeding trial was conducted to determine the optimal dietary vitamin C requirement and its effects on serum enzymes activities and bacterial resistance in the juvenile yellow drum Nibea albiflora (initial weight 33.2 ± 0.10 g). Six practical diets were formulated containing vitamin C 2.1, 45.3, 89.6, 132.4, 178.6 and 547.1 mg kg^?1 diet supplied as l ‐ascorbyl‐2‐monophosphate. The fish fed 547.1 mg kg^?1 diet showed a significantly higher survival than that fed 2.1 mg kg^?1 diet. The weight gains and specific growth rate of the fish fed 2.1 mg kg^?1 diet were significantly lower than those of the fish fed 89.6–547.1 mg kg^?1 diets. The liver vitamin C concentration firstly increased with increasing dietary vitamin C supply from 2.1 to 178.6 mg kg^?1 diet and then stabilized. The serum superoxide dismutase activities of the fish fed 547.1 mg kg^?1 diet were significantly lower than those of the fish fed 2.1–89.6 mg kg^?1 diet. The fish fed 2.1 mg kg^?1 diet had a significantly higher alkaline phosphatase activity than those in the other groups except the 45.3 mg kg^?1 group. Fish that received diets containing vitamin C at 547.1 mg kg^?1 had significantly higher nitro blue tetrazolium and lysozyme activity, and fish that received diets containing vitamin C at 45.3–547.1 mg kg^?1 exhibited resistance against Vibrio alginolyticus infection. The dietary vitamin C requirement of the juvenile yellow drum was established based on broken‐line model of weight gain to be 142.2 mg l ‐ascorbyl‐2‐monophosphate kg^?1 diet.     

Effects of dietary L‐carnitine level on growth performance,body composition and antioxidant status in beluga (Huso huso L. 1758)

A 17‐week feeding trial was carried out to evaluate the effects of dietary L‐carnitine level in beluga, Huso huso. A total of fish averaging 1247 ± 15.6 g (mean ± SD) were randomly distributed into 18 fibreglass tanks, and each tank holding 10 fish was then randomly assigned to one of three replicates of six diets with 50, 150, 350, 650, 950 and 1250 mg L‐carnitine kg^?1 diet. At the end of 17 weeks of feeding trial, average weight gain (WG), feed efficiency (FE), protein efficiency ratio (PER) and condition factor (CF) of fish fed 350 mg kg^?1 diet were significantly (P < 0.05) higher than those of fish fed 50, 150, 950 and 1250 mg kg^?1 diets. WG, FE, PER and CF of beluga fed 650 mg kg^?1 diet were also significantly higher than those of fish fed 50, 950 and 1250 mg kg^?1 diets. Whole body and muscle protein were significantly improved by the elevation of dietary L‐carnitine level up to 350 mg kg^?1. Liver superoxide dismutase and glutathione peroxidase activities of fish fed 350 and 650 mg kg^?1 diets were significantly higher than those of fish fed 50, 950 and 1250 mg kg^?1 diets. The dietary L‐carnitine level of 350–650 mg kg^?1 diet could improve growth performance, feed utilization, protein‐sparing effects of lipid, antioxidant defence system and reproductive success. Polynomial regression of WG suggested that the optimum dietary L‐carnitine level was 480 mg kg^?1 diet. Therefore, these results may indicate that the optimum dietary L‐carnitine could be higher than 350 but <650 mg kg^?1 diet in beluga reared in intensive culture conditions.     

Dietary lysine requirement of juvenile gilthead seabream Sparus aurata L.*

The dietary lysine requirement of juvenile gilthead seabream was determined by the growth response of duplicate groups of fish (3.5 g initial weight) fed on six isonitrogenous (427 g kg^?1) and isolipidic (135 g kg^?1) diets containing graded levels of crystalline l ‐lysine HCl, with dietary lysine content ranging from 36.3 to 79.7 g kg^?1 of protein. The final indispensable amino acid profile of the diets except for lysine was formulated so as to resemble that of wild seabream whole body. Except for the reduced growth performance of fish groups fed the lysine‐deficient diets no other deficiency signs were apparent. Survival observed throughout the feeding period of 6 weeks was excellent. Weight gain (in %), specific growth rate, feed efficiency and daily protein deposition (DPD) were significantly improved in response to the increasing levels of dietary lysine up to 52.7 g kg^?1 of protein and remained nearly constant thereafter. Whole‐body protein content followed a similar pattern as growth parameters in relation to dietary lysine level. Non‐linear regression analysis of DPD against dietary lysine level using the four‐parameter saturation kinetic model indicated a lysine requirement of 50.4 g kg^?1 of protein for this species to support growth.     

Oxidative effect of l -carnitine on energy metabolism in diploid and triploid rainbow trout ( Oncorhynchus mykiss ): impact on metabolites

l-Carnitine plays a key role in the regulation of energy metabolism and growth in fish. The aim of this study was to determine the effects of dietary l-carnitine levels on carnitine homeostasis and energy metabolism in diploid and triploid trout (Oncorhynchus mykiss). Diploid and triploid trout (29.5 ± 0.6 and 31.8 ± 0.5 g, respectively) were fed with three diets supplemented with 15, 200 or 530 mg l-carnitine/kg, respectively, for 56 days. Compared to 15 mg l-carnitine, total carnitine content increased in the liver by 182% in diploid and by 154% in triploid trout fed 530 mg l-carnitine. In muscle, total carnitine content increased by 60 and 73% in diploid and by 34 and 75% in triploid with the increase in dietary l-carnitine levels. l-Carnitine caused no significant change in the plasma concentrations of metabolites like proteins (ammonia, urea and protein), lipid (triglycerides), carbohydrate (glucose, lactate) and enzyme activities (lipase, lactate, alanine transaminase, lactic acid dehydrogenase). Triploid trout had significantly lower plasma ammonia (P = 0.003), lipase (P = 0.005) and triiodothyronine (T3; P = 0.003) levels than diploid trout. In conclusion, ploidy significantly affected the energy metabolism in rainbow trout, dietary l-carnitine levels altered the l-carnitine homeostasis, but not influence nutritional metabolism.     

Optimizing zinc supplementation levels of rainbow trout (Oncorhynchus mykiss) fed practical type fishmeal‐ and plant‐based diets

Rainbow trout (23.1 ± 0.4 g) were fed either a fishmeal‐ or plant‐based diet supplemented with various levels of zinc (0, 15, 30, 60 or 120 mg kg^?1) for 12 weeks. Trout fed the fishmeal diet had significantly higher weight gain than with the plant‐based diet. Zinc supplementation in the fishmeal diet had no effect on growth performance, suggesting that additional dietary supplementation of zinc is not required. However, in trout fed the plant‐based diet, growth increased significantly up to 30 mg kg^?1 zinc after which growth was not affected. Trout fed the plant‐based diet containing no zinc exhibited severe growth retardation, and in fish fed the 0 and 15 mg kg^?1 zinc diets, cataracts were present. Use of broken‐line quadratic modelling suggests that dietary supplementation of zinc needed to prevent deficiency and promote adequate growth in rainbow trout fed the plant‐based diet in this study was 30.1 mg kg^?1 (80 mg kg^?1 total dietary zinc). This is higher than the NRC (2011, Nutrient Requirements of Fish and Shrimp) dietary recommended level of 15 mg kg^?1 for rainbow trout. Following the NRC recommendation could lead to zinc deficiency in rainbow trout fed a plant‐based diet.     

Effect of dietary arachidonic acid levels on growth performance,hepatic fatty acid profile,intermediary metabolism and antioxidant responses for juvenile Synechogobius hasta

An 8‐week feeding experiment was conducted to determine the effect of dietary arachidonic acid (ARA) levels on growth performance, hepatic intermediary metabolism and antioxidant responses for juvenile Synechogobius hasta. Five isonitrogenous and isolipidic diets were formulated with arachidonic oil (containing 400 g ARA kg^?1) at inclusion levels of 0, 2, 4, 8 and 16 g kg^?1 to replace corn oil. Dietary ARA levels were 0.6, 8.6, 16.7, 32.7 and 64.8 g kg^?1 total fatty acids (FAs), respectively. Fish fed the 8.6–32.7 g ARA kg^?1 total FAs showed the highest weight gain, specific growth rate (SGR) and feed intake. By contrast, feed conversion ratio was the lowest for fish fed the 8.6–32.7 g ARA kg^?1 total FAs. Increasing ARA and total n‐6 fatty acid contents and declining linoleic acid content in liver were observed in fish fed the diet containing increasing dietary ARA levels. As a consequence, ∑n‐6/∑n‐3 ratios increased with increasing dietary ARA levels. Dietary ARA levels significantly influenced several enzymatic activities involved in hepatic intermediary metabolism, such as succinate dehydrogenase, lactate dehydrogenase, lipoprotein lipase and hepatic lipase. Superoxide dismutase activity increased with increasing dietary ARA levels. Glutathione peroxidase and catalase activities and malondialdehyde levels in liver tended to increase with increasing dietary ARA levels from 0.6 to 32.7 g ARA kg^?1 total FAs then declined when dietary ARA levels further increased to 64.8 g ARA kg^?1 total FAs. Broken‐line regression analysis of SGR against dietary ARA level indicated that optimal dietary ARA requirement for juvenile S. hasta was 10.74 g kg^?1 total FAs.     

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.     

Dietary tryptophan requirement of hybrid striped bass (Morone chrysops × M. saxatilis)

Requirements for six of the 10 indispensable amino acids (IAA) have not been quantified for hybrid striped bass. In this study, we estimate the requirement for l ‐tryptophan by nonlinear regression analysis of several growth indicators. Fifteen isocaloric, isonitrogenous diets were formulated to contain 1.0, 1.3, 1.6, 1.9, 2.2, 2.5, 2.8, 3.1, 3.4, 3.7, 4.0, 5.0, 6.0, 10.0 or 14 g l ‐tryptophan kg^?1 diet and fed to reciprocal cross hybrid striped bass for 7 weeks. After 5 weeks, survival of fish receiving the basal diet was 40% while surviving fish fed this diet were in poor health. Survival of fish receiving 1.3 g Trp kg^?1 diet declined to 70% by termination of the trial. Survival in other treatments was 100%. Fish weight gain was 100% or greater for fish receiving 1.6 g Trp kg^?1 diet or more. Hepatosomatic index, muscle ratio and intraperitoneal fat (IPF) ratio also responded to dietary tryptophan concentration. IPF was inversely related to dietary tryptophan concentration. The dietary tryptophan requirement was estimated to be between 2.1 and 2.5 g kg^?1 diet (6–7 g kg^?1 protein), depending on response variable, using four‐ and five‐parameter saturation kinetics models. These findings will increase the precision of diets formulated for hybrid striped bass.     

Effect of dietary l‐methionine concentrations on growth performance,serum immune and antioxidative responses of juvenile Nile tilapia,Oreochromis niloticus

An 8‐week feeding trial was conducted to determine the effect of dietary l ‐methionine supplementation on growth performance, serum immune and antioxidative responses of juvenile Nile tilapia Oreochromis niloticus. Six iso‐nitrogenous (282 g kg^?1 crude protein) diets were formulated to contain graded levels of methionine (0.32%, 0.54%, 0.75%, 0.93%, 1.14% and 1.33% of dry weight) at a constant dietary cysteine level of 0.83 g kg^?1. Each diet was randomly assigned to triplicate groups of 25 juvenile fish (2.3 ± 0.0 g), which were fed three times daily (8:30, 12:30 and 17:00 hours). The results showed that growth performance and feed utilization significantly improved when dietary methionine levels increased (P < 0.05). Using quadratic regression analysis of weight gain against dietary methionine levels indicated that, the optimal dietary methionine requirement for maximum growth of juvenile tilapia was 9.12 g kg^?1 of the dry diet in the presence of 0.83 g kg^?1 cystine. In addition, maximum C4 content and lysozyme activity were observed in fish fed 7.50 g kg^?1 methionine diet; maximum C3 content and superoxide dismutase activity were obtained in fish fed 9.30 g kg^?1 dietary methionine level. While there was no significant difference in serum glutathione peroxidase activity among all methionine supplemented treatments (P > 0.05). Methionine supplementation decreased malondialdehyde content in serum significantly (P < 0.05) when compared with the control diet, while there was no significant difference among supplemented treatments. These data suggested that l ‐methionine affected antioxidant status and promoted serum immune response in juvenile Nile tilapia, and at non‐stressed status, the requirement of dietary methionine in maintaining normal immunity and physiology is lower than that for maximum growth.     

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (4.20 ± 0.05 cm; 0.60 ± 0.02 g) was determined by conducting a 8‐week feeding trial with casein–gelatine‐based diets (400 g kg^?1 crude protein; 17.90 kJ g^?1, gross energy), containing crystalline amino acids with graded levels of l ‐arginine (10, 12.5, 15, 17.5, 20 and 22.5 g kg^?1, dry diet). Fish were randomly stocked, in triplicate groups, in 55‐L indoor polyvinyl flow through circular tanks and fed experimental diets at 5% of their body weight divided into two feedings at 08.00 and 16.00 hours. Live weight gain (321%) and feed conversion ratio (FCR 1.40) were significantly (P < 0.05) higher in fish fed diet containing 17.5 g kg^?1dietary arginine compared with other diets. Second‐degree polynomial regression analysis of live weight gain, FCR and protein efficiency ratio data indicated requirements for dietary arginine at 18.7, 18.4 and 18.3 g kg^?1 of the dry diet, respectively. Maximum carcass protein, and minimum moisture and fat contents were noticed at the requirement level. Carcass ash content remained insignificantly different among the treatments except at 17.5 g kg^?1 dietary arginine showing significantly higher ash content. Based on the above results, it is recommended that the diet for fingerling C. mrigala should contain arginine at 18.4 g kg^?1, dry diet, corresponding to 46 g kg^?1 dietary protein for optimum growth and efficient feed utilization.     

Optimal dietary methionine requirement of juvenile Chinese sucker,Myxocyprinus asiaticus

A total of 630 juvenile Chinese sucker, with an average initial weight of 1.72 ± 0.05 g, were fed seven diets for 56 days to study the effect of dietary methionine levels on growth, feed utilization, body composition and haematological parameters on juvenile Chinese sucker. Diet 1 using fish meal as the sole protein source and diets 2–7 using fish meal and fermented soybean meal as intact protein sources supplemented with crystalline amino acids contained six levels of l ‐methionine ranging from 6.4 to 18.9 g kg^?1 of dry diet at a constant dietary cystine level of 3.7 g kg^?1. Each diet was randomly assigned to three aquaria. Results indicated that the highest weight gain, specific growth rate (SGR), feed efficiency ratio, protein efficiency ratio and protein productive value occurred at 13.9 g methionine kg^?1 diet among the methionine supplemented dietary groups, beyond which they showed declining tendency. The whole body and muscle protein contents of juvenile Chinese sucker were positively correlated with dietary methionine level, while muscle lipid content was negatively correlated with it. The total essential amino acids content of muscle was increased significantly with increasing dietary methionine level from 6.4 to 13.9 g kg^?1 (P < 0.05). Apparent digestibility coefficients of dietary protein were significantly affected by dietary treatments. Serum protein, cholesterol and triacylglycerol increased with increasing dietary methionine levels, but showed a relatively lower value for fish fed the 18.9 g methionine kg^?1 diet. Quadratic regression analysis of SGR against dietary methionine level indicated that optimal dietary methionine requirement for juvenile Chinese sucker was 14.1 g kg^?1 of the diet in the presence of 3.7 g kg^?1 cystine (corresponding to 32.0 g kg^?1 of dietary protein on a dry‐weight basis).     

Dietary histidine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week growth trial was conducted to determine the dietary histidine requirement of the Indian major carp, Cirrhinus mrigala fingerling (length 4.22 ± 0.45 cm; weight 0.61 ± 0.08 g; n = 40). Isonitrogenous (400 g kg^?1 crude protein) and isoenergetic (17.90 kJ g^?1 gross energy) diets with graded levels of l ‐histidine (2.5, 5.0, 7.5, 10.0, 12.5 and 15.0 g kg^?1 dry diet) were formulated using casein and gelatin as a source of intact protein, supplemented with l ‐crystalline amino acids. Twenty fish were randomly stocked in 70‐L indoor polyvinyl circular fish tank (water volume 55‐L, water exchange rate 1–1.5 L min^?1) and fed experimental diets at the rate of 5% of their body weight/day divided over two feedings at 08:00 and 16:00 h. Maximum live weight gain (295%), best feed conversion ratio (FCR) (1.48) and protein efficiency ratio (PER) (1.69) occurred at 7.5 g kg^?1 of dietary histidine level. When live weight gain, FCR and PER data were analysed using second‐degree polynomial regression, the break points indicated histidine requirements at 9.4, 8.6 and 8.5 g kg^?1 of dry diet respectively. Significantly (P < 0.05) higher whole body protein and low moisture values were recorded at 7.5 g kg^?1 histidine level. Body fat increased significantly (P < 0.05) with increasing histidine levels. However, at 7.5 and 10 g kg^?1 histidine diets body fat did not differ (P > 0.05) to each other. Ash content of fish fed diets containing various levels of histidine did not differ except at 2.5 and 5.0 g kg^?1 inclusion levels where significantly (P < 0.05) higher ash was recorded. Protein deposition was also found to be significantly (P < 0.05) higher in the 7.5 g kg^?1 histidine diet. Based on the polynomial regression analysis of FCR and PER data, it is recommended that the diet for fingerling C. mrigala should contain histidine at 8.5 g kg^?1 of dry diet, corresponding to 21.25 g kg^?1 of dietary protein for optimum growth and efficient utilization of feed.     

Efficacy of an equal blend of canola oil and poultry fat as an alternate dietary lipid source for Atlantic salmon (Salmo salar L.) in sea water. I: effects on growth performance, and whole body and fillet proximate and lipid composition

This study assessed the suitability and cost efficacy of an equal blend of canola oil (CO) and poultry fat (PF) as a supplemental dietary lipid source for juvenile Atlantic salmon. Quadruplicate groups of Atlantic salmon (～400 g) held in 4000 L outdoor fibreglass tanks supplied with running (35–40 L min^?1), aerated (dissolved oxygen, 7.88–10.4 mg L^?1), ambient temperature (8.6–10.9°C) sea water (salinity, 26–35 g L^?1) were fed twice daily to satiation one of three extruded dry pelleted diets of equivalent protein (488–493 g kg^?1 dry matter) and lipid (267–274 g kg^?1 dry matter) content for 84 days. The diets were identical in composition except for the supplemental lipid (234.7 g kg^?1) source viz., 100% anchovy oil (AO; diet COPF‐0), 70.2% AO and 29.8% CO and PF (diet COPF‐30), and 40.3% AO and 59.7% CO and PF (diet COPF‐60). Atlantic salmon growth rate, feed intake, feed efficiency, protein and gross energy utilization, percent survival and whole body and fillet proximate compositions were not affected by diet treatment. Cost per kilogram weight gain was about 10% less for fish fed diet COPF‐60 than for diet COPF‐0. Percentages of saturated fatty acids in dietary and fillet lipids varied narrowly. Moreover, percentages of 18:1n‐9, monounsaturated fatty acids, 18:2n‐6, n‐6 fatty acids, 18:3n‐3, and ratios of n‐6 to n‐3 fatty acids in the flesh lipids were directly related to the dietary level of CO and PF whereas 22:6n‐3, the total of 20:5n‐3 (eicosapentaenoic acid; EPA) and 22:6n‐3 (docosahexaenoic acid; DHA), and n‐3 fatty acids revealed the opposite trend. Percentages of 22:6n‐3, EPA and DHA, and n‐3 fatty acids were significantly depressed in fish fed diet COPF‐60 versus diet COPF‐0. We conclude that a 1:1 blend of CO and PF is an excellent cost‐effective dietary source of supplemental lipid for Atlantic salmon in sea water.