The effect of dipeptide,Lys‐Gly,supplemented diets on digestive tract histology in juvenile yellow perch (Perca flavescens)

The aim of the present study was to evaluate the effects of three formulated diets: wheat‐gluten‐protein‐based diets supplemented with Lys‐Gly dipeptide (LG) or free lysine and glycine (FL), a control diet without lysine supplementation (C) and commercial starter Bio Oregon (BO) for on the growth and digestive system morphology of yellow perch. After 48 days of experimental feeding, fish fed LG diet showed the highest body mass. Fish fed LG diet showed the highest number of gastrin/cholecystokinin positive cells and the lowest number of CD3‐positive cells. The brush border of anterior intestine was the most PepT1 immunopositive in fish fed LG diet, the weakest in fish fed C diet. The largest hepatocytes were observed in fish fed BO, while the smallest in those fed FL diet, the difference being statistically significant. Relative hepatocyte cytoplasm volume occupied by lipids was higher in fish fed BO and FL compared to those fed C and LG. Number of proliferating cell nuclear antigen‐positive hepatocyte nuclei did not significantly differ among experimental groups. These results indicate that wheat‐gluten‐protein‐based diets supplemented with dipeptide Lys‐Gly (LG) were appropriate for yellow perch.     

The influence of feeding diets containing wheat gluten supplemented with dipeptides or free amino acids on structure and development of the skeletal muscle of carp (Cyprinus carpio)

A 28-day feeding trial was conducted to evaluate the influence of feeding diets containing plant protein wheat gluten supplemented with dipeptides or free amino acids on structure and development of the skeletal muscles of carp (Cyprinus carpio). Common carp fingerlings (1 month old) having an of average weight of 0.07 ± 0.02 g and total length of 17.79 ± 1.79 mm were fed three formulated diets—wheat gluten protein-based diets supplemented with Lys–Gly dipeptide (PP), free lysine and glycine (AA), control diet without lysine supplemented (CON)—and two other diets: restrictive diet—frozen zooplankton (Z) and commercial diet Aglo Norse (AN). After 28 days of experimental feeding, statistically significant higher survival was observed among fish fed AN and Z diets (99.5 ± 1.0 %; P ≤ 0.05). The feeding AN diet has had also a positive influence on weight and growth rate as well as on development and growth of skeletal muscles. Furthermore, carps fed AN diet had the largest area of red and white muscle as compared with the other feeding groups, and the differences were statistically significant (P ≤ 0.05). The increase in the number of proliferating cells (proliferating cell nuclear antigen) was observed on the last day of the experiment among carps fed PP, AA and CON. Moreover, fish fed PP significantly had the greatest number of MyoD- and myogenin-positive nucleus (P ≤ 0.05). Among the experimental diets based on wheat gluten, a positive impact on structure and development of muscles has been observed in carps fed PP diet.     

The effect of lysine‐supplemented wheat gluten‐based diet on yellow perch Perca flavescens (Mitchill) performance

Formulation of nutritionally complete and cost efficient diets for yellow perch (Perca flavescens) is a prerequisite for successful intensive culture of this species. One of the objectives of this study was to determine the optimum diet for the grow‐out phase of juvenile yellow perch. Fish at the size of 12.9 ± 4 g were individually marked with passive integrated transponders (PIT)‐tags and randomly distributed into six 400 L tanks, 45 fish per tank. This experiment included lysine‐deficient [(?) Lys] and lysine‐supplemented [(+) Lys] wheat‐gluten‐based diets in triplicate groups. Our experiment showed that the mean weight of fish fed (+) Lys diet (83.9 ± 1.5 g) was significantly larger than fish fed (?) Lys diet (68.6 ± 5.2 g) (P < 0.05). This experiment also showed that the blood plasma concentration of free lysine in (+) Lys group was significantly higher than in (?) Lys group (P < 0.05) and the same trend appeared also in methionine concentrations. The concentration of Lys in deficient group of fish, 3 h after a meal was lower compared with levels of Lys 24 h post‐feeding (P < 0.05). Lysine deficiency in diet resulted in significantly higher level of serine, and a similar trend occurred in small and large fish.     

Requirements of vitamin C (l-ascorbyl-2-sulphate and l-ascorbyl-2-polyphosphate) and its effects on non-specific immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to determine two vitamin C derivatives, l ‐ascorbyl‐2‐sulphate (C2S) and l ‐ascorbyl‐2‐polyphosphate (C2PP), to satisfy vitamin C requirement and to test their effects on the non‐specific humoral immune responses of juvenile grouper, Epinephelus malabaricus. C2S and C2PP were each supplemented at 20, 50, 80, 150, 250 and 400 mg kg^?1 diet in the semi‐purified basal diet providing of 7, 16, 28, 55, 86, 142 mg ascorbic acid (AA) equivalent of C2S kg^?1 diet and 4, 9, 15, 31, 49, 75 mg AA equivalent of C2PP kg^?1 diet, respectively. Basal diet without AA supplemented was included as a control. Each diet was fed to triplicate groups of grouper (mean initial weight: 6.69 ± 0.07 g). Fish fed diets with ≥28 mg AA equivalent of C2S or ≥4 mg AA equivalent of C2PP kg^?1 had significantly (P < 0.05) greater weight gain (WG) than fish fed the unsupplemented control diet. Liver AA concentrations were higher in fish fed diets with ≥16 mg AA equivalent of C2S or ≥9 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Alternative pathway of complement activation (ACP) was higher in fish fed diets with ≥55 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Lysozyme activity was higher in fish fed ≥86 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 46.2 mg AA equivalent of C2S kg^?1 diet and 17.8 mg AA equivalent of C2PP kg^?1 diet, and it also indicated that C2S is approximately 39% as effective as C2PP in meeting the vitamin C requirement for grouper. The data suggest that both C2S and C2PP supplementation support non‐specific immune responses of grouper.     

Free- and peptide-based dietary arginine supplementation for the South American fish pacu (Piaractus mesopotamicus)

Arginine was hypothesized to be a model compound in the present study on molecular forms of indispensable amino acid (IAA) dietary supplementation. Juvenile South American pacu (Piaractus mesopotamicus) were fed diets containing arginine in a protein base (casein‐wheat gluten or casein‐gelatin), or the casein‐wheat gluten base supplemented with dipeptide or free arginine at two levels (5 and 10 g kg^?1). Growth and protein efficiency ratios were significantly affected by diets, but not by arginine molecular form. Three free dispensable amino acids (DAA) and four IAA in plasma were affected by diet, but plasma arginine concentrations did not differ. Plasma urea concentrations, being very low in the pacu, and hepatic arginase activities, were not affected by diet (P = 0.10–0.11), but together with plasma ornithine, mirrored the growth data. Molecular form of arginine supplementation, free or dipeptide, significantly changed several free IAA (Phe, Leu, Ile, His) and urea, with a higher mean plasma concentration in dipeptide fed fish. The dietary treatments, or molecular form of the arginine supplementation, did not change proximate composition, except that calcium levels decreased with higher dietary arginine supplementation level. The present study indicates that dipeptides can provide IAA to pacu, and that arginine supplemented in this form is utilized as efficiently as in free form.     

Partial substitution of fish meal and full-fat soya bean meal with wheat gluten and influence of lysine supplementation in diets for rainbow trout, Oncorhynchus mykiss (Walbaum)

A 65-day study was undertaken to evaluate the utilization of lysine-supplemented wheat gluten meal as a protein source for rainbow trout, Oncorhynchus mykiss (Walbaum). The performance of the trout fed wheat gluten was compared to that of trout where protein was principally supplied using fish meal and full-fat soyabean meal. Six isonitrogenous diets (approximately 45% crude protein) were formulated. Thus, the reference diet (FS) was based on fish meal and full-fat soya bean meal while a further five diets were formulated with a mixture of wheat gluten and wheat middlings used to isonitrogenously replace approximately 75% and 57% of the full-fat soya and fish meal respectively. Crystalline l -lysine representing 0.00%, 0.29%, 0.58%, 0.87% and 1.16% of the complete diet was then added to the wheat-gluten-based diets. Optimal performance in terms of weight gain and apparent net protein utilization was achieved by the fish fed a wheat-gluten-based diet supplemented with lysine (0.58%) yielding a digestible lysine level of 1.9% of the complete feed. Within the range of supplements provided, lysine digestibility was high. However, at the highest levels of supplemental lysine the relationship between uptake and supplement level was not linear. Additionally, arginine digestibility was not affected by the level of lysine supplementation. However, amongst the wheat-gluten-based diets, optimal performance was associated with a lysine: arginine ratio of 1.1: 1. Despite a lysine: arginine ratio of approximately 1.1: 1, the overall performance of the fish fed the fish-meal-soya-based reference diet was poorer than expected. The results are discussed with respect to optimal patterns of gross and digestible amino acids, lysine-arginine antagonism and the possible relationship between antinutritional factors and the poor performance of the fish fed the fish-meal-soya-based reference diet.     

Growth performance and feed utilization of Nile tilapia Oreochromis niloticus (Linnaeus, 1758) and tilapia galilae Sarotherodon galilaeus (Linnaeus, 1758) fingerlings fed plant protein-based diets

This study was designed to determine the effect of complete substitution of fish meal (FM) by three plant protein sources including extruded soybean meal (SBM), extruded full‐fat soybean (FFSB) and corn gluten meal (CGM) on growth and feed utilization of Nile tilapia Oreochromis niloticus and tilapia galilae Sarothrodon galilaeus. Four isonitrogenous of crude protein (ca. 28.0%) and isocaloric (ca. 19 MJ kg⁻¹) experimental diets were formulated. The control diet (diet 1) was prepared with FM as the main protein sources. Diets 2–4, each FM control diet, were completely substituted with SBM (diet 2), FFSB (diet 3) and CGM (diet 4). l ‐lysine and dl ‐methionine were added to plant protein diets to cover the nutritional requirements of tilapia. Each treatment was allocated to three net pens and fed for 17 weeks. Nile tilapia fed the control diet showed significantly higher (P≤0.05) values for final body weight (FBW), feed intake (FI), weight gain (WG) and specific growth rate (SGR), whereas fish fed the diet with CGM achieved the lowest values. Tilapia galilae fed SBM diet recorded the highest (P≤0.05) values for growth performance. Better feed conversion ratio (FCR) for both Oreochromis niloticus and Sarothrodon galilaeus was observed when fish were fed SBM diet, whereas the worse FCR was recorded for FFSB diet. Feed utilization parameters including protein productive value (PPV), fat retention (FR) and energy retention (ER) showed significant differences (P≤0.05) for both the species fed different dietary protein sources. The present results suggest that, for Nile tilapia, both SBM and FFSB supplemented with dl ‐methionine and l ‐lysine can completely replace dietary FM. Meanwhile, S. galilaeus fed SBM diet exhibited comparable growth and feed utilization with those fish fed a fish‐meal‐based diet.     

The Effects of Plant-protein-based Diets Supplemented with Yucca on Growth, Digestibility, and Chemical Composition of Nile Tilapia (Oreochromis niloticus, L) Fingerlings

Replacement of fish meal (FM) protein with dehulled and solvent‐extracted plant by‐products, soybean meal, cottonseed meal, sunflower meal, and linseed meal was tested in diets for juvenile Nile tilapia, Oreochromis niloticus. Fish averaging (±SD) 14.2 ± 2.9 g were divided into 18 groups and fed for 6 mo on pelleted feed containing each of the plant protein meal supplemented with Yucca schidigera powder extract at 750 mg/kg. Methionine (1%) and lysine (0.5%) were added to each diet except the control diet (FMC), while diet FMC + Y was supplemented with yucca only. Three groups of fish were fed each of six isonitrogenous (25% crude protein) and isocaloric (4.3 kcal/g) diets replacing 100% of FM protein and performance compared against a nutritionally balanced control and a commercial tilapia feed. After 6 mo of feeding, the fish fed plant protein diets supplemented with yucca exhibited growth performance not differing significantly from that of fish fed FMC + Y, while differing significantly from the control FMC and diet linseed meal (LSM). The highest apparent protein digestibility coefficient was observed for diets treated with yucca, which was significantly higher than that observed for the control diet FMC. No significant differences were found in whole‐body moisture of fish fed different experimental diets. An increase in the whole‐body protein content was observed in fish fed diets supplemented with yucca, which was significantly different from that of the diet FMC. The whole‐body fat content of Nile tilapia was low and showed significant differences among treatments supplemented with yucca compared with the control diet FMC. The whole‐body energy content showed the same trend as whole‐body fat content.     

Senegalese sole juveniles (Solea senegalensis Kaup, 1858) grow equally well on diets devoid of fish meal provided the dietary amino acids are balanced

A growth trial was conducted with the aim of developing a practical, environmentally sustainable and cost-effective plant-protein-based diet for Senegalese sole juveniles without compromising growth or protein deposition. A reference diet contained fish meal as the main protein source (F45, containing 37% fish meal and 45% of the dietary protein being from marine sources) and four plant-protein diets either containing 5% (F15) or no fishmeal (F5). The F15 diet also contained 5% hydrolysed protein (CPSP) and 5% squid meal while the F5 contained 2% CPSP and 3% squid meal. The rest of the protein derived from a blend of soybean meal, corn and wheat gluten. Thus, the plant-protein fraction in these test diets ranged from 70–80%. The plant-based diets were supplemented either with a mixture of all indispensable amino acids (IAA) (F15 + IAA, F5 + IAA) or only with lysine (F15 + Lys, F5 + Lys). Triplicate groups of 18 fish of 9.5 g were fed by automatic feeders that distributed 8 meals over 24 h, for a period of 12 weeks.At the end of the experiment mean body weight more than tripled in all treatments (32.8 to 42.7 g). Sole fed with either F15 + Lys (33.3 g) or F5 + Lys (32.8 g) grew significantly less than all the other treatments. Protein productive value (PPV) was significantly lower in sole fed with F5 + Lys (0.30) than in all other treatments (0.34 to 0.37) while F5 + Lys (0.23) energy productive value (EPV) just differed significantly from fish fed with F45 (0.38). Moreover, protein gain (P gain) was significantly higher in sole fed with diet supplemented with all IAA (F15 + IAA and F5 + IAA) than those fed with diet with only lysine added (F15 + Lys and F5 + Lys) and a strong positive correlation between P gain and methionine (Met) intake was found (P = 0.00002).Dietary fish meal protein can be totally replaced by a mixture of plant-protein sources without any adverse effects on growth, feed or protein utilization provided the dietary amino acids are balanced. The test diet F5 + IAA seems a good alternative to use in Senegalese sole production when the main objective is to substitute an increasingly scarce ingredient – fish meal – without compromising fish performance. Moreover, for large-scale production feeds the F15 + Lys putatively supplemented with Met, Thr and Arg would be recommended.     

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.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

The effects of dietary protein levels with amino acid supplementation on the growth performance,haematological profile and histology of meagre (Argyrosomus regius) in two different size classes

A 12‐week feeding trial was conducted to estimate the effects of supplemented amino acids (lysine and methionine) with different dietary protein levels on growth, haematology and liver histology in meagre (Argyrosomus regius) at two different sizes: fingerling (15.2 g) and juvenile (44.4 g). Six practical diets were prepared, and three of these practical diets were formulated with 40 (P40), 44 (P44) and 48% (P48) crude protein without supplementation of amino acids. Other three diets having same protein levels were supplemented with lysine and methionine (P40AA, P44AA and P48AA) to bring these amino acids level up to that estimated to be at the least in the 48% protein diet. Final mean weight (FMW) and specific growth rate (SGR) of P48AA in fingerlings were higher than that of 40AA. According to FMW and SGR of juveniles, there was interaction found between dietary protein levels and the supplementation of amino acids. The FMW and SGR of meagre fed P44AA were higher than that of fish fed the other diets, except fish fed the P48. Economic profit index, packed cell volume and haemoglobin for juvenile meagre were significantly improved with the dietary protein level and amino acid supplementation. The liver tissues of the P48 group in both sizes displayed better structure compared with the other groups. In conclusion, dietary crude protein content of practical diets for juvenile meagre could be reduced from 48% to 44% by adding essential amino acids with an enhancement in growth, health and economic profit.     

Bio‐Ag reutilization of distiller's dried grains with solubles (DDGS) as a substrate for black soldier fly larvae,Hermetia illucens,along with poultry by‐product meal and soybean meal,as total replacement of fish meal in diets for Nile tilapia,Oreochromis niloticus

A feeding trial was conducted in a closed system with Nile tilapia, Oreochromis niloticus, juveniles (mean initial weight, 2.66 g) to examine total replacement of menhaden fish meal (FM) with distiller's dried grains with solubles (DDGS), which had been used as substrate for the production of black soldier fly larvae, Hermetia illucens, in combination with soybean meal (SBM) and poultry by‐product meal (PBM), with or without supplementation of the amino acids (AA) DL‐methionine (Met), L‐lysine (Lys) and a commercial non‐amylaceous polysaccharide enzyme (Enz) product. Fish were fed seven isoenergetic [available energy (AE) = 4.0 kcal g^?1 of diet] and isonitrogenous (350 g kg^?1 protein as‐fed basis) practical diets formulated with equivalent digestible protein levels. Diet 1 was formulated to be similar to a commercial, high‐quality, tilapia diet containing 200 g kg^?1 FM. Diets 2–5 were formulated as a 2 × 2 factorial to replace FM with similar contributions from DDGS (45%), PBM (25%) and SBM (2.1–2.9%), but to differ in supplementation of AA and/or Enz preparation. Diets 6 and 7 were formulated to investigate the effects of a 2/3 and 1/3 reduction, respectively, in DDGS contribution to the replacement protein mix, with concomitant increases in SBM, with respect to diet 3, and were balanced with Lys and Met. After 6 weeks, growth responses were slightly attenuated (P ≤ 0.05) and average daily intake (ADI) and feed conversion ratio (FCR) were slightly higher in tilapia fed DDGS diets 2–5 compared to those of fish fed the FM control diet 1. Growth responses were not significantly affected by the presence or absence of AA or Enz (diets 2–5), or the level of DDGS (diets 3, 7 and 6). Whole‐body proximate composition was not different among treatments. Amino acid profiles of fish fed DDGS diets were not significantly different from those of fish fed the FM control. Evidence of interaction between AA and Enz supplementation was detected in whole‐body amino acid concentrations such that AA content was higher with AA or Enz addition alone, but lower when both were added to the diet. Results suggest that DDGS replacement of FM in tilapia diets can be substantial when diets are formulated on a digestible protein basis and DDGS is combined with highly digestible animal (PBM) and plant proteins (SBM).     

Effect of fish meal replacement by plant protein blend on amino acid concentration,transportation and metabolism in juvenile turbot (Scophthalmus maximus L.)

The objective of the study was to evaluate the effects of incorporating plant protein blend in juvenile turbot (Scophthalmus maximus L.) diet on free amino acid (AA) concentration and the expression of genes related to peptide and AA transporters, key enzymes of AA metabolism and AA response (AAR) pathway. Fish were fed diets with fish meal (FM), or 400 g/kg FM replacement by plant protein blend for 9 weeks. Compared with the FM diet, PP40 diet did not affect plasma essential amino acid (EAA) concentration or AA metabolic enzymes gene in intestine, while it significantly upregulated all the detected peptide and neutral AA transporters gene. Results in muscle indicated that PP40 diet led to a great reduction of EAA concentrations and mRNA abundance of two kinds of AA transporters (SNAT2 and b^0,+AT), while it greatly increased the gene expression of L‐type and T‐type AA transporters (LAT2 and TAT1) and the enzymes of AA catabolism (BCKDH‐E2) and anabolism (asparagine synthetase). In addition, the expression of genes related to AAR pathway were all greatly stimulated by PP40 diet in muscle. Our results provide a molecular explanation for the change of tissues AA concentrations caused by plant protein in turbot, which maybe applicable for general carnivorous fish.     

Effect of dietary phosphorus supplementation on utilization of algae in the grow‐out diet of Nile tilapia Oreochromis niloticus

This research involved testing the replacement of corn gluten protein concentrate in the diet of 1.5 g Nile tilapia with biofuel algae‐based or algae supplemented with phosphorus diets. Experimental diets were formulated to replace 50% of the gluten meal protein in the control diet with biofuel algae protein (Alga50) or Spirulina (Spirulina50) protein. In addition, dicalcium phosphate (P) was added to the Alga50 at a concentration of 3.8% or 7.74% to mitigate aluminium (Al) toxicity and designated as Alga50 + P, and Alga50 + PP respectively. After 9 weeks of the feeding experiment, fish fed diets supplemented with P, Alga50 + P and Alga50 + PP differed significantly (P < 0.05) with respect to the feed conversion and protein deposition from the control. The mineral composition of the fish body showed a decrease in levels of Al and iron (Fe) due to the P supplements. The mineral composition of the faeces indicated that Al and Fe were excreted in the faeces and were not accumulated in the fish body. The effect of dicalcium P supplementation on the neutralization of Al in the diet resulted in improved fish growth and histological integrity of the gastrointestinal tract.     

Do plant‐based diets for Atlantic cod (Gadus morhua L.) need additions of crystalline lysine or methionine?

Atlantic cod (Gadus morhua), initial weight 15 g, were fed ten experimental diets for 15 weeks. The diets were based on a mixture of plant proteins (PP) and fish meal (FM), where PP constituted 65% of dietary protein. PP mixtures were chosen to reach as low levels of lysine and methionine as possible. The diets were supplemented with increasing amounts of lysine (19.2–31.9 g kg^?1 diet) or methionine (9.4–12.3 g kg^?1 diet), in a regression design. No growth difference among diet groups was found in the plant‐based diets. Increased dietary lysine resulted in decreased liver size, plasma triacylglycerol concentration (TAG) and lipid productive value (LPV). Methionine additions did not result in changed Hepatosomatic index (HSI), LPV or plasma TAG. Feed conversion ratio (FCR) and protein utilization were neither affected by lysine nor methionine. Plasma and muscle concentrations of free lysine and methionine correlated with dietary levels 5‐h post feeding. Overall conclusion was that cod maintain growth rates in plant‐based diets if dietary protein was high, without additional supplements of crystalline lysine or methionine. Lysine intake significantly influenced lipid metabolism, showing the necessity to add lysine in plant protein‐based diets to hinder increased lipid deposition. No such effects were found because of lack of methionine additions.     

Dietary lysine requirement of large yellow croaker (Pseudosciaena crocea,Richardson 1846) larvae

A 30‐day feeding experiment was conducted to estimate the lysine requirement of large yellow croaker larvae (2.75 ± 0.11 mg). Six isonitrogenous (509.5–519.7 g kg^?1 crude protein) and isoenergetic (22.3–22.5 kJ g^?1 energy) microdiets containing graded levels of lysine·HCl ranging from 24.8 to 41.0 g kg^?1 diet in placement of glycine and glutamic acid were formulated. Mixture of crystalline amino acids (MAA) was supplemented to simulate the amino acid (AA) profiles of whole body of this larva, except for lysine. The MAA and supplemented lysine for each diet were coated with tripalmitin. Triplicate groups of 3000 fish were fed to apparent satiation by hand eight times per day. The results showed that specific growth rate (SGR), survival, body composition and the specific activity of digestive enzymes were significantly affected by dietary lysine levels (P<0.05). The optimal dietary lysine requirements estimated by second‐order polynomial model based on SGR and survival were 33.7 (65.5 g kg^?1 dietary protein) and 33.4 (64.9 g kg^?1 dietary protein) g kg^?1 dry diet respectively. The estimated requirements for the other essential AAs were calculated by A/E ratios of whole body AA profile of this larva based on lysine requirement.     

Fish protein hydrolysate affected amino acid absorption and related gene expressions of IGF‐1/AKT pathways in turbot (Scophthalmus maximus)

The effects of fish protein hydrolysate (FPH) on growth, peptide and amino acid (AA) transporters, postprandial free AA and related gene expression of IGF‐1/AKT pathway were evaluated in turbot (Scophthalmus maximus). Three diets were formulated to contain the same low level of fishmeal; meanwhile 0, 45 and 180 g/kg FPH were, respectively, supplemented to the FF (FPH‐free), FL (FPH‐Low) and FH (FPH‐High) diets. Fish fed the FH diet improved the growth compared with the other groups. For peptide and AA transporters, PepT1, B⁰AT1, CAT1 and PAT1 mRNA levels in proximal or distal intestine decreased in fish fed the FH diet. The concentration of free total essential AAs in serum was higher in the FH treatment than that in the FF treatment at 2 and 6 hr after feeding. For IGF‐1/AKT pathway in muscle, IGF‐1, 4E‐BP1 and FoxO1 mRNA levels were the highest in the FH group, whereas IGF‐1R mRNA levels were the highest expression level in the FF group. In conclusion, down‐regulated AAs transport, alleviated the delayed postprandial peak of serum‐free AAs and increased muscle protein synthesis were observed to improve the growth when turbot was fed high FPH level diets containing a high plant protein.     

Effects of dietary fishmeal replacement with Spirulina platensis on the growth,feed utilization,digestion and physiological parameters in juvenile gibel carp (Carassis auratus gibelio var. CAS III)

A 60‐day growth experiment was conducted to investigate the effect of dietary replacement of fishmeal by Spirulina platensis on growth performance, digestion and physiological parameters in juvenile gibel carp (5.0 ± 1.0 g). Four diets were formulated to replace 0 (SP0), 50% (SP50), 75% (SP75) and 100% (SP100) of dietary fishmeal protein by S. platensis respectively. Other two diets based on SP100 were supplemented with 2% dicalcium phosphate (SP100 + P) or 0.28% coated lysine (SP100 + Lys). The results showed that no significant differences of specific growth rate (SGR), feed efficiency (FE), feeding rate (FR) and protein efficiency ratio (PER) were observed between SP0 group and the replacement groups (SP50, SP75 and SP100). FE and PER of fish‐fed SP100 + Lys diet were significantly higher than the fish‐fed SP100 diet (p < .05). Apparent digestibility coefficients (ADCs) of dry matter, protein, energy and phosphorus increased significantly with the increasing S. platensis inclusion. Meanwhile, fish from SP100 + P or SP100 + Lys group had higher ADCs of nutrients compared to the SP100 group (p < .05). Compared with SP0 diet, fish‐fed S. platensis diets had higher activities of plasma superoxide dismutase, hepatopancreas total antioxidant capacity, plasma alkaline phosphatase and plasma lysozyme, as well as the lower content of plasma malondialdehyde. The present results indicated that dietary fishmeal could be fully replaced by S. platensis without any negative effects on growth performance of gibel carp. Supplementation of lysine in the S. platensis replacement diet could further improve the feed utilization of gibel carp.     

Effects of Neutral Phytase Supplementation on Biochemical Parameters in Grass Carp,Ctenopharyngodon idellus,and Gibel Carp,Carassius auratus gibelio,Fed Different Levels of Monocalcium Phosphate

Two 8‐wk studies were conducted to evaluate the effects of neutral phytase supplementation on hemato‐biochemical status, liver biochemical parameter, and intestinal digestive enzyme activity of grass carp, Ctenopharyngodon idellus, and gibel carp, Carassius auratus gibelio, fed with different levels of monocalcium phosphate (MCP). The control diet was prepared with 2% MCP but without phytase (P2.0). The other three experimental diets were prepared with the addition of 1.5, 1.0, and 0.5% MCP, respectively, when supplemented with 500 U/kg neutral phytase in each diet and designated as PP1.5, PP1.0, and PP0.5, respectively. The results indicated that the serum alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate transaminase (AST) activities, as well as the albumin (ALB) content were increased in grass carp (P < 0.05) and gibel carp (P > 0.05) fed with phytase‐supplemented diets. Meanwhile, the serum cholesterol, high‐density lipoprotein, and total protein contents of the two species of fish were increased in comparison to the control. In addition, dietary phytase inclusion did not significantly affect hepatic ALP, ALT, and AST activities in the two species of carp fed with different levels of MCP. Amylase activity increased in foregut and hindgut of both species when fed with the phytase‐supplemented diets while lipase activity was reduced in the foregut and hindgut in both fish. This study suggests that neutral phytase supplementation increases serum ALP, ALT, and AST activities but does not notably affect these enzyme activities in the liver of the two species of carp when fed different levels of MCP. On the other hand, amylase activity increased while lipase activity was reduced in the intestine of the species of carp fed with phytase‐supplemented diets.