Fast growth, protein turnover and costs of protein metabolism in yolk-sac larvae of the African catfish (Clarias gariepinus)

In fish larvae the costs of rapid growth may be accommodated by a decrease in the rate of protein turnover or by a reduction in the costs of protein synthesis. Protein growth, synthesis and degradation were measured in yolk-sac larvae of Clarias gariepinus and the costs of protein synthesis and protein growth were estimated. Growth rates were over 100% protein weight day^-1. Protein synthesis retention efficiency (retained protein per unit of synthesis) was estimated to be 69.6%, a value comparable to that of larger fish. The larvae used 43% of their oxygen consumption for protein synthesis. Nevertheless, protein synthesis costs were close to theoretical minima. Therefore, the high growth rates of catfish yolk-sac larvae seem to be possible through minimisation of the costs of protein synthesis. These low costs are associated with high rates of protein synthesis (138%protein weight day^-1), and elevated RNA concentrations (107 µg RNA mg^-1protein), which together suggest very high RNA efficiencies (12.9 g protein synthesized g^-1RNA day^-1).     

Effects of insecticide Thiodan® on the morphology and quantification of ovarian follicles in lambaris Astyanax bimaculatus (Linnaeus, 1758) in different treatments

Thiodan^® is an insecticide which can cause morphological changes in fish tissues, dependent on the concentration. To investigate possible morphological changes, this study evaluated the number and diameter of ovarian follicles in lambaris Astyanax bimaculatus exposed to Thiodan^® for 96 h in different treatments at three sub‐lethal concentrations: 1.15, 2.3 and 5.6 μg L^?1 and a control group without Thiodan^®. Females exposed to Thiodan^® were divided into four treatments: (i) without acclimation and without food, (ii) without acclimation and with food, (iii) with acclimation for 10 days and without food; (iv) with acclimation for 10 days and with food. In this study, it was noted that the action of Thiodan^® at sub‐lethal concentrations in a static system did not affect the morphology of follicular development. However, the treatments with acclimation showed lower numbers of primary and secondary follicles when compared to fish from treatments without acclimation. For the groups exposed to Thiodan^®, a greater number of atretic follicles were observed compared to the control groups. The follicular diameter of secondary follicles in fish exposed to Thiodan^® in the treatment with acclimation and with food was highest in the control group (P < 0.05). Furthermore, the mean gonadosomatic (GSI) and hepatosomatic (HSI) indices of treatments with acclimation were lower than those for treatments without acclimation. These data suggest that the fish may be being affected by the pesticide to which the fish were submitted, and may cause impairment to the follicular development.     

Maturation of the pancreatic and intestinal digestive functions in sea bass (Dicentrarchus labrax): effect of weaning with different protein sources

The maturation of the digestive functions in sea bass (Dicentrarchus labrax) larvae was evaluated by the enzymatic profile of pancreas and intestine brush border membranes. Sea bass larvae were weaned at day 25 with three simplified diets different by their protein nature: 100% fish meal (FP), 100% casein mixture (CP) and 50% fish meal-50% casein mixture (CFP). The casein mixture contained 35% of hydrolysate. The control group was fed live preys. The specific activity of amylase decreased with age irrespectively of the diets whereas the specific activity of trypsin was enhanced. The casein mixture reduced pancreatic secretion in amylase and trypsin. The CFP group differed from the other groups fed on compound diets, exhibiting as soon as day 32 high activities of brush border enzymes, similar to controls. This sharp increase between day 25 and 32 appeared to be crucial for larval survival. The addition of a protein hydrolysate in a weaning diet seems to facilitate this maturation process.     

Positive feedback control by the gonads on gonadotropin (GTH) and gonadoliberin (GnRH) levels in experimentally matured female silver eels,Anguilla anguilla

Treatment of sham-operated female silver eels with carp pituitary extract stimulated ovarian development and induced increases in pituitary gonadotropin (GTH) and gonadoliberin (GnRH) contents. Both effects of carp pituitary extract were abolished in ovariectomized eels, indicating the involvement of the gonads. Endogenous sexual steroids, the secretion of which was increased during sexual maturation, should be responsible for the stimulation of GTH and GnRH levels. Ovariectomy itself had no significant effect on pituitary GTH and GnRH contents, reflecting the fact that, at the silver stage, sexual steroid levels are too low to exert any significant effect on pituitary GTH and GnRH. The positive feedback control exerted by the gonads on GTH and GnRH levels during sexual maturation, in the eel as well as in some other teleosts, would produce an amplification of the pubertal stimulation of the hypothalamo-pituitary-gonadal axis.     

Water quality affects SDH activity, protein content and RNA:DNA ratios in fish (Catla catla, Labeo rohita and Oreochromis mossambicus) raised in ponds of a sewage-fed fish farm

The spatial and seasonal variability of the respiratory enzyme succinate dehydrogenase and the protein content were examined in different tissues of fish cultured in three ponds along the effluent gradient of a sewage-fed fish farm. Indian major carp Catla catla (150-230 g) and Labeo rohita (60-190 g) cultured in both the middle and last points of the sewage effluent (stocking pond 1) and (stocking pond 4) and Oreochromis mossambicus (50-160 g), a naturally growing fish of the inlet (facultative pond) and the out let of the sewage effluent (stocking pond 4) were procured every month during the period of January-December, 2005 and were subjected to determination of succinate dehydrogenase activity, total protein, DNA and RNA contents from gill, liver and muscle tissue respectively. The SDH activity of all three test fishes (Catla catla, Labeo rohita and Oreochromis mossambicus) was reduced significantly (ANOVA; P < 0.05) when cultured in SP-4 compared to SP-1 in the case of Catla catla and Labeo rohita and in facultative pond in the case of Oreochromis mossambicus.Conspicuous differences in the SDH activity of fish between the last and first stocking pond or the facultative pond were clearly due to the result of the differences in water quality. There was a direct relationship between SDH activity in gill tissue of any of the fish investigated and ammonia-N concentration of water or water pH. This shows that the respiratory activity of these fishes was strongly affected by the ammonia and pH of water. In other words, this suggests that as the distance from the point source increases, there was a substantial improvement of water quality in the ponds located along the sewage effluent gradient. Evidently, there is a progressive pattern of growth, survival and physiological health of fish and abundance of favorable diversity of food organisms with rich biodiversity.     

The effects of elevated winter temperature and sub-lethal pollutants (low pH, elevated ammonia) on protein turnover in the gill and liver of rainbow trout (Oncorhynchus mykiss)

Appetite, growth, and protein turnover (synthesis, growth and degradation) of liver and gills were measured in juvenile rainbow trout (Oncorhynchus mykiss) fed to satiation, and exposed for 90 days to elevated winter temperatures (+2 °C above ambient) and either low pH (5.2) in softwater or 70 M total ammonia (TAmm) in hardwater. All fish increased in weight during the experiments, but those exposed to +2°C grew significantly more than those at ambient temperature due to a stimulation of appetite. During the relatively constant temperature of the first 75 days, +2 °C caused a significant increase in the rates of protein synthesis and degradation in the liver of hardwater-acclimated fish, as a result of an increase in RNA translational efficiency (KRNA). The elevated temperature also induced an increase in gill protein synthesis in softwater-acclimated fish but in this case the underlying mechanism was an increase in Cs, the capacity for protein synthesis (RNA:protein) rather than in KRNA. The addition of 70 M TAmm had no effect on protein turnover in either liver or gills of hardwater-acclimated fish. Low pH inhibited protein growth in the liver of softwater-acclimated fish at day 90 under both temperature regimes. This inhibition was effected via a decrease in protein synthesis at control temperature but via an increase in protein degradation when the fish were exposed to both low pH and +2 °C. From these results we conclude that a simulated global warming scenario has potentially beneficial rather than detrimental effects on protein turnover and growth of freshwater fish during winter.     

A comparison of RNA concentrations and ornithine decarboxylase activity in Atlantic salmon (Salmo salar) muscle tissue,with respect to specific growth rates and diel variations

RNA concentrations and enzyme activities are often used as indices of recent growth in fish, but few studies have used both methods to assess the same fish. This study measured RNA concentrations and ornithine decarboxylase (ODC) activity in muscle tissue of juvenile Atlantic salmon (Salmo salar) to compare their usefulness for reflecting specific growth rates, and to determine whether either growth index was influenced by diel variations or time of feeding. Three groups (n = 54 in total) were fed 1.5% of body weight in commercial pellets in four feedings per day. One group was fed only in the morning (0830–1230h), one in the afternoon (1430–1830h), and one in the morning and afternoon (0830–1830h). At the end of ten days, fish were sampled at three times (0130h, 1030h, 1630h) over a single 24h period. Correlations to specific growth rate were slightly higher for RNA concentrations than for ODC activity, but both were highly significant. RNA and ODC activity were also correlated to each other. These results suggest that RNA concentration and ODC activity, taken together, can be used to monitor changes in both the numbers and activity of ribosomes. For RNA concentrations, there was no evidence of an effect of diel variations or the time of feeding. For ODC activity, a significant diel effect (all feed schedules combined) was detected if one non-growing fish was excluded from the analysis; activity of the enzyme was slightly higher in the sample taken at night (0130h) than in the two daytime samples. Contribution no. 8, Catamaran Brook Habitat Research Project     

Effects of starvation on RNA : DNA ratio, glycogen content, and C : N ratio in columellar muscle of the Japanese turban shell Turbo (Batillus) cornutus (Gastropoda)

ABSTRACT: The validity of using the RNA : DNA ratio, glycogen content, and C : N ratio in the columellar muscle as indicators of the nutritional condition in the Japanese turban shell Turbo cornutus was examined. These biochemical indices were compared between fed and starved animals. The fed animals were fed brown algae to satiation for 108 days as a control, whereas the starved animals were not fed for the first 75 days and then fed for next 33 days. All three indices declined during the starvation period, and the values for the starved group were significantly lower than those for the fed group during days 20–75. At day 108, after the re-feeding period, the indices for the starved group were found to have increased. Among the indices, only the RNA : DNA ratio for the starved males responded rapidly to starved conditions and became significantly lower even at day 4, and only the RNA : DNA ratio for the starved males and females recovered to those levels of the fed animals at day 108. The results indicated that the RNA : DNA ratio is the most rapid indicator of nutritional stress among the three indices. However, it is recommended that glycogen content and C : N ratio be used in addition to the RNA : DNA ratio for monitoring the health of T. cornutus as the RNA : DNA ratio showed large variations.     

Effect of combined fish meal : soybean meal ratio,vitamin C and fish oil supplementations in diet on the growth and reproduction of red swamp crayfish,Procambarus clarkii (Crustacea: Decapoda)

Fish meal (FM), vitamin C (VC) and fish oil (FO) play important roles in the growth and reproduction in aquatic animals. To determine the optimum dietary combinations of FM:SM (soybean meal) ratio, VC and FO supplementations for the growth and reproduction of female crayfish Procambarus clarkii, we used the L9 (3³) orthogonal array, which arranged three factors varied at three levels (FM:SM at 1:3, 1:2 and 1:1.5; VC supplementation at 0.02%, 0.04% and 0.06%; FO inclusion at 1.90%, 2.85% and 3.80%). Range analysis showed that the highest weight gain (WG) of the crayfish occurred at FM:SM of 1:1.5, VC supplementation of 0.04% and FO inclusion of 1.90%, and the maximum gonadosomatic index (GSI), egg laying amount (ELA) and spawning rate (SR) were obtained at FM:SM 1:1.5, VC supplementation 0.06% and FO inclusion 3.80%. Analysis of variance indicated that all the three factors had no significant effects on WG. Fish oil supplementation significantly affected GSI, ELA and SR (P<0.05), while FM:SM did not significantly influence any of these indicators. Taking the cost into consideration, we suggest that a dietary FM:SM of 1:3, VC supplementation 0.06% and FO inclusion 3.80% might be the optimal combinations for the reproduction of P. clarkii.     

Aspects of gametogenesis, oocyte morphology and maturation of the lugworm Arenicola marina (Annelida: Polychaeta) in relation to commercialised procedures to extend the breeding season

Environmental manipulation is a common method of extending the spawning season of aquaculture species including the polychaete worm Arenicola marina. Temperature synchronises autumn spawning populations and so its manipulation was used to advance and delay spawning. Females were exposed to a minimum period of 3 weeks at 5 °C in conjunction with the injection of prostomial homogenate to induce spawning up to 4 weeks prior to the natural spawning date. We also maintained individuals at 15–17 °C starting 4 weeks prior to, and then continuing after the natural spawning date, delaying spawning for up to 4 months. Both sexes can be manipulated, but males suffered higher mortalities and a greater rate of spontaneous spawning within the tanks. In ‘advanced’ females, mean oocyte diameters (measured in September, one month prior to spawning) were significantly larger and more homogenous compared to ambient individuals, whilst ‘delayed’ females produced a second cohort of oocytes approximately 8 weeks into the treatment. Delaying and advancing spawning induced significant changes in the ultrastructural morphology of prophase and metaphase oocytes, and delayed prophase oocytes showed a significant increase in the number with cracks on the surface of the vitelline membrane. Although, SDS-PAGE and Western blots confirmed that Maturation Promoting Factor (MPF) activity was not different from ambient controls, there were significant changes in MPF activity levels (measured by histone kinase activity) in manipulated oocytes. A. marina has the plasticity for spawning to be delayed and advanced by a number of months; this is essential for the continued development of aquaculture of this species. However, the maturational ability of the oocytes is compromised and this may have significant implications for production and quality of the offspring from manipulated individuals.     

Identification and characterization of a phospholipase A1 activity type three secreted protein,PP_ExoU from Pseudomonas plecoglossicida NB2011, the causative agent of visceral granulomas disease in large yellow croaker (Larimichthys crocea)

Pseudomonas plecoglossicida NB2011, the causative agent of visceral granulomas disease in farmed Larimichthys crocea in China, encodes a predicted type three effector PP_ExoU, a homolog of the cytotoxin ExoU of Pseudomonas aeruginosa. In this study, secretion of PP_ExoU was tested in various broth, the protein was expressed with the pET30a prokaryotic system, the phospholipase A (PLA) activity of the recombinant protein was determined with fluorogenic phospholipid substrates, fusion expression with green fluorescent protein in transfected HeLa cells was investigated, and the lactate dehydrogenase (LDH) level was measured. The results showed the protein was type three secreted in several media; the recombinant protein displayed significant PLA1 activity with ubiquitin. Fluorescence was observed on the cell membrane and scattered in the cytoplasm of HeLa cells expressing catalytic wild‐type PP_ExoU, blebbing and stretching developed in the cell membranes indicating of membrane damage. Fluorescence scattered in the cytoplasm of cells expressing the catalytic inactive protein. A significant LDH level was detected in HeLa cells expressing wild‐type PP_exoU, but not in the Ser/Asp‐mutated protein, suggestion mutation of predicted catalytic residues abolished the PLA activity. This is the first report on the function of a secreted type three protein from P. plecoglossicida.     

1H NMR‐based metabolomics studies on the effect of size‐fractionated fish protein hydrolysate,fish meal and plant protein in diet for juvenile turbot (Scophthalmus maximus L.)

This study was designed to evaluate changes in the metabolic profile of liver and muscle of turbot (Scophthalmus maximus L.) fed fishmeal‐based diet, diets containing size‐fractionated fish protein hydrolysate and plant protein‐based diet using ¹H NMR‐based metabolomics approach combined with the growth. Fish protein hydrolysate (FPH) was obtained by enzymatic treatment, permeate fraction was obtained as UF by ultrafiltered step, and retentate fraction was retained as RF. FM diet contained fish meal used as a single protein source. Four other diets (PP, UF, FPH and RF) contained 180 g kg^?1 diet fish meal. 54, 55 and 55 g kg^?1 dry diet UF, FPH and RF were supplemented to UF, FPH and RF diets. All diets were formulated to be isolipidic and isonitrogenous fed to five triplicate groups of turbot (16.05 ± 0.03 g) for 68 days. O‐PLS‐DA in FM versus UF, FM versus FPH, FM versus RF and FM versus PP resulted in a reliable model for muscle and liver tissue, while O‐PLS‐DA in UF versus FPH and UF versus RF only showed metabolites changes in liver tissue. Results indicated that metabolite changes among the different treatments were consistent with the growth tendency.     

Effects of feeding practical diets containing different protein levels, with or without fish meal, on growth, survival, body composition and processing traits of male and female Australian red claw crayfish (Cherax quadricarinatus) grown in ponds

Four practical diets containing 2% of crude protein (CP) (180 and 280 g kg^?1), with or without menhaden fish meal (FM), were fed to Australian red claw Cherax quadricarinatus during a 97‐day feeding trial. Growth, survival, body composition and processing traits of pond‐cultured red claw were determined. Juvenile red claw (mean individual weight of 5.75 ± 3.3 g) were randomly stocked into twelve 0.04‐ha ponds at a rate of 1000 per pond (25 000 ha^?1), and each diet was fed to three ponds. At harvest, the final mean weight of red claw fed Diet 4 (0 g kg^?1 FM and 280 g kg^?1 CP) and Diet 3 (113 g kg^?1 FM and 280 g kg^?1 CP) was significantly (P < 0.05) higher (62.4 and 58.5 g, respectively) than red claw fed Diet 1 (73 g kg^?1 FM and 180 g kg^?1 CP; 51.7 g) and Diet 2 (0 g kg^?1 FM and 180 g kg^?1 CP; 53.0 g). Red claw fed diets containing 280 g kg^?1 CP, with or without FM, had significantly higher percent weight gain (894 and 959%, respectively) compared to red claw fed 180 g kg^?1 CP, with or without FM (778 and 799%, respectively). Feed conversion ratio, percent survival, and total yield among treatments, which averaged 3.55, 65.2%, and 724 kg ha^?1 overall, were not significantly different. Results from this study indicate that pond‐cultured red claw stocked at 25 000 ha^?1 can be fed a practical diet containing 280 g kg^?1 CP with 0 g kg^?1 FM if a combination of plant‐protein ingredients (soybean meal, distillers’ dried grains with solubles, and milo) is added; however, if the percentage of dietary protein level is 180 g kg^?1, growth is reduced even if FM is added at 73 g kg^?1 of the diet. Use of diets containing plant‐protein sources may help reduce diet costs to producers and thereby, increase profits.     

A review of sawfishes (Pristidae) in the Arabian region: diversity,distribution, and functional extinction of large and historically abundant marine vertebrates

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Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

The effect of time‐dependent protein restriction on growth factors,nonspecific immunity,body composition,fatty acids and amino acids in the Siberian sturgeon (Acipenser baerii)

The effect of time‐dependent protein restriction (PR) and refeeding were investigated on growth, body composition, fatty acids (FA), amino acids (AA) and nonspecific immune functions in the juvenile Siberian sturgeon (Acipenser baeri; 75 ± 5 g). Test diets included: T1 (control) and T2 fish fed diets containing 400 and 300 g/kg protein respectively during the whole experimental period, T3 (every other day) and T4 (every other week) fish were fed diets containing 300 g/kg protein (PR) and refeeding with diet containing 400 g/kg protein respectively, and T5 fish fed a diet containing 300 g/kg protein for 3 weeks and a diet containing 400 g/kg protein for 5 weeks, were fed to the fish for 56 days by visual satiation. Unlike ration T2 and T5, feeding treatments of T3 and T4 showed an increase in fish growth and body composition (p < 0.05) and they were very close to the control group (p > 0.05). Regarding the fatty acid profile, although the percentage of highly unsaturated fatty acids (HUFA) and polyunsaturated fatty acids (PUFA) declined significantly in the T2 and T5 groups, the saturated fatty acid (SFA) and monounsaturated fatty acids (MUFA) indicated no change among the treatments. In addition, the T2 and T5 groups demonstrated a reduction in essential amino acids (EAA) with a simultaneous increase in nonessential amino acids (NEAAs), which was significant from that of controls and other groups (p < 0.05). In terms of nonspecific immune parameters (serum lysozyme activity and alternate complement activity [ACH50]), treatment 3 has an appropriate result that did not have a significant difference with the control group (p > 0.05), but there was significant difference with other groups (p < 0.05). As a result, the T3 treatment can be used in sturgeon aquaculture practically without any negative impacts on biological or physiological indices.     

Replacement of fishmeal by soya protein concentrate in the diets of Totoaba macdonaldi (Gilbert, 1890) juveniles: effect on the growth performance,in vitro digestibility,digestive enzymes and the haematological and biochemistry parameters

The potential of soya protein concentrate (SPC) as an alternative protein source in diets for Totoaba macdonaldi juveniles was evaluated. Seven isonitrogenous and isolipidic diets were formulated containing 15–100% SPC (SPC15, SPC30, SPC45, SPC60, SPC75, SPC90 and SPC100) to replace fishmeal (FM‐protein), and a FM‐protein‐based diet without SPC was used as a reference diet (RD). Each diet was randomly assigned to triplicate groups of 20 totoaba (50 ± 1.0 g) and was fed twice daily to apparent satiation. After 60 days of experiment, effects on totoaba were evaluated. Growth performance in fish fed RD to SPC45 was similar. The maximum FM‐protein replacement for weight gain (g kg ABW^?1 day^?1) was estimated to be 34.17% using a broken‐line model. In vitro digestibility of fish fed RD, SPC15 and SPC30 was similar. Trypsin activity was higher in fish fed the RD (9.38 ± 0.52 mU × 10^?3 mg protein^?1. Chymotrypsin activity was similar in fish fed RD, SPC15 and SPC30. Activity of alkaline protease and phosphatase was similar in fish fed RD, SPC15 and SPC30. Red blood cells (RBC) were the highest in fish fed RD to SPC60. Fish fed the RD, SPC15 and SPC30 present the highest haematocrit (HT) and haemoglobin (HB) content. Diets SPC90 and SPC100 presented similar MCV and MCHC. Total protein (TP) profile overall decreased in fish fed SPC90 and SPC100, pointing to nutritional hypoproteinemia due to deficient digestion and absorption. These results indicate that SPC‐based diets could be used efficiently by totoaba with FM‐protein replacement of less than 45%.     

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.