Indispensable amino acid concentrations decrease in tissues of stomachless fish,common carp in response to free amino acid- or peptide-based diets

Summary. The premise that free amino acid or dipeptide based diets will resolve the nutritional inadequacy of formulated feeds for larval and juvenile fish and improve utilization of nitrogen in comparison to protein-based diets was tested in stomachless fish, common carp (Cyprinus carpio L.) larvae. We examined the postprandial whole body free amino acid (FAA) pool in fish that were offered a FAA mixture based diet for the duration of 2 or 4 weeks. We found that the total amount and all indispensable amino acids concentrations in the whole body decreased after a meal. We then fed juvenile carp with dietary amino acids provided in the FAA, dipeptide (PP), or protein (live feed organisms; brine shrimp Artemia salina nauplii, AS) forms. Histidine concentrations in the whole fish body increased in all dietary groups after feeding whereas all other indispensable amino acids decreased in FAA and PP groups in comparison to the AS group. Taurine appears to be the major osmotic pressure balancing free amino acid in larval freshwater fish which may indicate a conditional requirement. We present the first evidence in larval fish that in response to synthetic FAA and PP diets, the whole body indispensable free AA concentrations decreased after feeding. This study shows that amino acids given entirely as FAA or PP cannot sustain stomachless larval fish growth, and may result in depletion of body indispensable AA and most of dispensable AA. The understanding of these responses will determine necessary changes in diet formulations that prevent accelerated excretion of amino acids without protein synthesis.     

Protein digestion, growth and food conversion in Atlantic salmon and Arctic charr with different trypsin-like isozyme patterns

Protein digestibility and food conversion were determined in groups of Atlantic salmon ( Salmo salar L.) and Arctic charr ( Salvelinus alpinus L.) with or without the trypsin-like isozyme TRP-2*92 . Determinations were made at two salinities, 0 and 27 ppt; at two temperatures, 6 and 10° C and on large (200 g) and small (30–90 g) fish. The overall digestibility of protein was found to be 81.4±0.2% and was unaffected by the presence of the isozyme or the other variables. The feed conversion ratio of Atlantic salmon possessing TRP-2*92 was observed to be significantly lower with significantly higher specific growth rate than in fish without this isozyme.     

Effects of dietary trimethylamine on free amino acid and nonprotein nitrogen levels in muscle of the guppy, Poecilia reticulata, in relation to seawater adaptation

Guppies Poecilia reticulata acclimated to 100% seawater (SW) had lower taurine and alanine levels in muscle than fish kept in freshwater (FW). The glycine level, in contrast, was higher in SW fish than in FW fish. Levels of other free amino acids (FAA) were comparatively low and little different between fish adapted in FW and in SW. In both FW and SW fish almost all of muscle FAA showed little difference in levels between fish kept on diets containing three different levels of trimethylamine (TMA) (0, 223, and 334 mumol TMA/kg dry weight of diet). Total FAA and nonprotein nitrogen levels in muscle were unaffected by the difference in either the diet species or the ambient salinities. Muscle trimethylamine oxide levels were higher in SW fish than in FW fish. In both salinities, muscle trimethylamine oxide levels in fish on the diets containing 223 and 334 mumol TMA/kg were slightly greater than the level in fish on the TMA-free diet.     

Effect of dietary conjugated linoleic acid (CLA) on lipid composition, metabolism and gene expression in Atlantic salmon (Salmo salar) tissues

Dietary conjugated linoleic acid (CLA) affects fat deposition and lipid metabolism in mammals, including livestock. To determine CLA effects in Atlantic salmon (Salmo salar), a major farmed fish species, fish were fed for 12 weeks on diets containing fish oil or fish oil with 2% and 4% CLA supplementation. Fatty acid composition of the tissues showed deposition of CLA with accumulation being 2 to 3 fold higher in muscle than in liver. CLA had no effect on feed conversion efficiency or growth of the fish but there was a decreased lipid content and increased protein content after 4% CLA feeding. Thus, the protein:lipid ratio in whole fish was increased in fish fed 4% CLA and triacylglycerol in liver was decreased. Liver beta-oxidation was increased whilst both red muscle beta-oxidation capacity and CPT1 activity was decreased by dietary CLA. Liver highly unsaturated fatty acid (HUFA) biosynthetic capacity was increased and the relative proportion of liver HUFA was marginally increased in salmon fed CLA. CLA had no effect on fatty acid Delta6 desaturase mRNA expression, but fatty acid elongase mRNA was increased in liver and intestine. In addition, the relative compositions of unsaturated and monounsaturated fatty acids changed after CLA feeding. CLA had no effect on PPARalpha or PPARgamma expression in liver or intestine, although PPARbeta2A expression was reduced in liver at 4% CLA feeding. CLA did not affect hepatic malic enzyme activity. Thus, overall, the effect of dietary CLA was to increase beta-oxidation in liver, to reduce levels of total body lipid and liver triacylglycerol, and to affect liver fatty acid composition, with increased elongase expression and HUFA biosynthetic capacity.     

Effect of starvation on free histidine and amino acids in white muscle of milkfish Chanos chanos

Milkfish (Chanos chanos) decreased their body weight from 47 to 28 g over the 60-day period of starvation. Starvation also resulted in the reduction of muscle lipid and protein, and hepatosomatic index. The predominant free amino acid (FAA) in white muscle of milkfish was histidine, followed by taurine and glycine. In the first 25 days of starvation, no significant change in histidine was found. After 40 days of starvation, however, the histidine concentration was significantly decreased by 46%, and remained unchanged thereafter. As compared to control group fish, the 60-day-starved fish possessed only half the amount of histidine. Taurine and glycine, on the other hand, showed no significant changes throughout starvation. Taurine became the most predominant in the FAA pool after 40 days of starvation, and the concentration of 60-day-starved fish was two times higher than that of control group fish without starvation. The ratios of histidine, taurine, and glycine to total FAAs remained approximately the same although the individual contributions varied considerably to the total FAAs during starvation. The results of this study suggested that a good strategy would be to keep taurine and glycine in milkfish muscle at relatively high levels for physiological function as histidine decreased drastically for energy source under conditions of food deprivation.     

Metabolic adjustments to varying protein intake in harbor seals (Phoca vitulina): evidence from serum free amino acids

Effects of varying dietary protein intake on serum free amino acid (FAA) concentrations were studied in harbor seals (Phoca vitulina) fed two different prey fish diets: either exclusively low-fat, high-protein walleye pollock (Theragra chalcogramma) or high-fat, relatively high-energy-density Pacific herring (Clupea pallasi). Significant differences in FAA concentrations and patterns were observed between the two diets. All essential amino acids (EAA), except methionine and phenylalanine, and two nonessential amino acids (NEAA), glycine and tyrosine, decreased when the diet was switched from herring to pollock and increased on switching back to herring. Both total EAA concentrations and EAA : NEAA ratios decreased with the elevated protein intake typical of a low-fat pollock diet, indicating an inverse correlation between EAA concentrations and dietary protein intake levels. We propose that differing dietary protein intake, caused by differences in macronutrient composition of the two prey fish species, induced a change in protein metabolism that was reflected in blood-circulating amino acids. These findings suggest that surveys of amino acid profiles may be useful to partially determine the protein metabolic status of harbor seals.     

Purine-induced expression of urate oxidase and enzyme activity in Atlantic salmon (Salmo salar). Cloning of urate oxidase liver cDNA from three teleost species and the African lungfish Protopterus annectens

The peroxisomal enzyme urate oxidase plays a pivotal role in the degradation of purines in both prokaryotes and eukaryotes. However, knowledge about the purine-induced expression of the encoding gene is lacking in vertebrates. These are the first published sequences of fish urate oxidase, which were predicted from PCR amplified liver cDNAs of Atlantic salmon (Salmo salar), Atlantic cod (Gadus morhua), Atlantic halibut (Hippoglossus hippoglossus) and African lungfish (Protopterus annectens). Sequence alignment of different vertebrate urate oxidases revealed amino acid substitutions of putative functional importance in the enzyme of chicken and lungfish. In the adult salmon, expression of urate oxidase mRNA predominated in liver, but was also identified in several nonhepatic organs including brain, but not in skeletal muscle and kidney. Juvenile salmon fed diets containing bacterial protein meal (BPM) rich in nucleic acids showed a significant increase in liver urate oxidase enzyme activity, and urea concentrations in plasma, muscle and liver were elevated. Whereas salmon fed the 18% BPM diet showed a nonsignificant increase in liver mRNA levels of urate oxidase compared with the 0% BPM-fed fish, no further increase in mRNA levels was found in fish receiving 36% BPM. The discrepancy between urate oxidase mRNA and enzyme activity was explained by rapid mRNA degradation or alternatively, post-translational control of the activity. Although variable plasma and liver levels of urate were detected, the substrate increased only slightly in 36% BPM-fed fish, indicating that the uricolytic pathway of Atlantic salmon is intimately regulated to handle high dietary purine levels.     

Effects of a high plant protein diet on the somatotropic system and cholecystokinin in Atlantic salmon (Salmo salar L.)

To investigate the endocrine signalling from dietary plant protein on somatotropic system and gastrointestinal hormone cholecystokinin (CCK), two iso-amino acid diets based on either high plant or high fish meal protein were fed to Atlantic salmon. Salmon with an average starting weight of 641 ± 23 g (N = 180), were fed a fish meal (FM) based diet (containing 40% FM) or diets mainly consisting of blended plant proteins (PP) containing only 13% marine protein, of which only 5% was FM for 3 months. mRNA levels of target genes GH, GH-R, IGF-I, IGF-II, IGFBP-1, IGF-IR in addition to CCK-L, were studied in brain, hepatic tissue and fast muscle, and circulating levels of IGF-I in plasma of Atlantic salmon were measured. We detected reduced feed intake resulting in lower growth, weight gain and muscle protein accretion in salmon fed plant protein compared to a diet based on fish meal. There were no significant effects on the regulation of the target genes in brain or in hepatic tissues, but a trend of down-regulation of IGF-I was detected in fast muscle. Lower feed intake, and therefore lower intake of the indispensable amino acids, may have resulted in lower pituitary GH and lower IGF-I mRNA levels in muscle tissues. This, together with higher protein catabolism, may be the main cause of the reduced growth of salmon fed plant protein diet. There were no signalling effects detected either by the minor differences of the diets on mRNA levels of GH, GH-R, IGF-IR, IGF-II, IGFBP-1, CCK or plasma protein IGF-I.     

Daily changes in parameters of energy metabolism in liver, white muscle, and gills of rainbow trout: dependence on feeding

We assessed the daily patterns of parameters involved in energy metabolism in liver, white muscle, and gills of rainbow trout. Where daily rhythms were found, we analyzed the potential influence of feeding. Immature rainbow trout were randomly distributed in 3 groups: fish fed for 7 days, fish fasted for 7 days, and fish fasted for 7 days and refed for 4 days. On sampling day, fish of fed and refed groups were fed at 11.00 h, and all fish were sampled from each treatment group using the following time schedule: 14.00, 18.00, 21.00, 00.00, 04.00, 07.00, 10.00 and 14.00 h. The results obtained from metabolic parameters can be grouped into four different categories, such as i) those displaying no daily changes in any group assessed in liver (acetoacetate and lactate levels), white muscle (protein levels, and low Km (glucose) hexokinase (HK) and HK-IV activities) and gills (protein levels), ii) those displaying no 24 h changes in fed fish but in refed or fasted fish in liver (glucose, glycogen, amino acid and protein levels, and HK-IV activity), white muscle (glycogen and amino acid levels) and gills (glucose levels), iii) those displaying 24 h changes that were apparently dependent on feeding since they disappear in fasted fish in liver (Low Km (glucose) HK, lactate dehydrogenase (LDH-O), glucose 6-phosphatase (G6Pase), fructose 1,6-bisphosphatase (FBPase) , alpha-glycerophosphate dehydrogenase (G3PDH), glutamate dehydrogenase (GDH) and aspartate aminotransferase (Asp-AT) activities), white muscle (glucose levels, and pyruvate kinase (PK), LDH-O, G3PDH and Asp-AT activities) and gills (glycogen and lactate levels, and Low Km (glucose) HK, HK-IV, LDH-O and Asp-AT activities), and iv) those parameters displaying 24 h changes apparently not dependent on feeding in liver (lactate levels and PK activity) and gills (amino acid levels, and PK and GDH activities). In general, most 24 h changes observed were dependent on feeding and can be also related to daily changes in activity.     

High activities of cathepsins B, D, H, and L in the white muscle of chum salmon in spawning migration

1. Activities of cathepsins, lysosomal hydrolytic enzymes and cysteine protease inhibitor in both the white and red muscles of chum salmon (Oncorhynchus keta) caught during spawning and feeding migrations were compared. 2. In the white muscle, cathepsins B, D, H and L activities were 3-7 times higher in the fish in spawning migration than those in feeding migration. However, in the red muscle, no such marked differences were observed between them. 3. Cysteine protease inhibitory activity and extractable protein content in the white muscle of the fish in spawning migration were about 40% lower than those in feeding migration. 4. The present study supports the conception that the cathepsins are related to protein catabolism of the fish during spawning migration.     

Influence of dietary non-essential amino acid profile on growth performance and amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.)

The quality of dietary protein is an important factor influencing the growth performance of fish. To evaluate the quality of protein, the variables commonly studied are the composition of the essential amino acids, the digestibility and the protein use efficiency. The goal of the present experiment was to test the effect of the dietary non-essential amino acid composition on the growth of Nile tilapia (Oreochromis niloticus). The fish were fed three purified diets differing only in their non-essential amino acid composition. The influence of the experimental diets on the growth performance, on the activity of enzymes involved in the amino acid metabolism, aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT), and on whole body delta(15)N values was investigated. Body mass, lipid, protein and energy gain differed significantly between the feeding groups. The activity of ASAT in the whole liver was significantly higher in fish with a positive protein balance compared to fish which lost protein. Whole body delta(15)N values of fish were negatively correlated with their body mass gain. Despite the poor utilisation of synthetic amino acids, the experiment indicates the importance of the dietary non-essential amino acid composition for the growth performance of fish. The study reveals the possibility to trace the utilisation of synthetic amino acids by determining the isotopic composition of dietary amino acids and tissues or whole bodies of animals.     

Non-essential amino acids play an important role in adaptation of the rat exocrine pancreas to high nitrogen feeding

We have previously demonstrated that feeding a diet with a high amino acid (60% AA diet) content, as a mixture simulating casein, induced pancreatic growth and pancreatic protease production in rats. In the present study, we examined the effects of an increasing dietary content of essential amino acids (EAA, x1 - x3 in exp. 1 and x1 - x3.3 in exp. 2) and non-essential amino acids (NEAA, x1 - x3 in exp. 1 and x1 - x5.2 in exp. 2) on pancreatic growth, amylase and protease adaptation using casein-type amino acid mixtures (exp. 1, basal diet; 20% AA diet) and egg white-type amino acid mixtures (exp. 2, basal diet; 12% AA diet). Pancreatic growth and trypsin activity were induced as the dietary content of NEAA was increased in experiments 1 and 2. Amylase activity in the pancreas was also induced as the dietary content of NEAA was increased, even with the decrease in dietary carbohydrate in experiment 2. The values of all pancreatic variables decreased with the increase in dietary EAA (x2 and x3) without an increase in NEAA. The changes in the pancreas were coincident with increases in plasma arginine and lysine concentrations and a decrease in the plasma alanine concentration. In rats fed a 60% AA diet (EAA and NEAA x3), in the case of which the EAA content was balanced with the NEAA content, pancreatic growth and protease production increased and reached maximum levels as the plasma amino acid concentrations decreased, except for alanine. These results show that NEAA, not EAA, are associated with induction of pancreatic growth and protease production upon feeding a diet with a high AA content, and that some metabolites may be involved in the induction process. The suppression of pancreatic growth and protease production in rats fed the high EAA diets without balanced NEAA may be associated with impairment of amino acid metabolism rather than the increments in the concentration of one or more essential amino acids. Our results also suggest that there is an unknown mechanism or unknown factors involved in regulating pancreatic amylase.     

Branched-chain amino acid supplementation during bed rest: effect on recovery.

Bed rest is associated with a loss of protein from the weight-bearing muscle. The objectives of this study are to determine whether increasing dietary branched-chain amino acids (BCAAs) during bed rest improves the anabolic response after bed rest. The study consisted of a 1-day ambulatory period, 14 days of bed rest, and a 4-day recovery period. During bed rest, dietary intake was supplemented with either 30 mmol/day each of glycine, serine, and alanine (group 1) or with 30 mmol/day each of the three BCAAs (group 2). Whole body protein synthesis was determined with U-(15)N-labeled amino acids, muscle, and selected plasma protein synthesis with l-[(2)H(5)]phenylalanine. Total glucose production and gluconeogenesis from alanine were determined with l-[U-(13)C(3)]alanine and [6,6-(2)H(2)]glucose. During bed rest, nitrogen (N) retention was greater with BCAA feeding (56 +/- 6 vs. 26 +/- 12 mg N. kg(-1). day(-1), P < 0.05). There was no effect of BCAA supplementation on either whole body, muscle, or plasma protein synthesis or the rate of 3-MeH excretion. Muscle tissue free amino acid concentrations were increased during bed rest with BCAA (0.214 +/- 0.066 vs. 0.088 +/- 0.12 nmol/mg protein, P < 0.05). Total glucose production and gluconeogenesis from alanine were unchanged with bed rest but were significantly reduced (P < 0.05) with the BCAA group in the recovery phase. In conclusion, the improved N retention during bed rest is due, at least in part, to accretion of amino acids in the tissue free amino acid pools. The amount accreted is not enough to impact protein kinetics in the recovery phase but does improve N retention by providing additional essential amino acids in the early recovery phase.     

Digestive enzymes and metabolic profile of Labeo rohita fingerlings fed diets with different crude protein levels

Labeo rohita, commonly called rohu is one of the most important fish species for aquaculture in India. Digestive enzyme response and metabolic profile of fingerling L. rohita to different dietary crude protein (CP) levels (viz. 25, 30, 35 and 40%) were studied in an attempt to optimize a practical diet formulation for this species. After 45 days of feeding, activity of digestive enzymes and metabolite concentrations were assayed. Amylase, lipase and alkaline phosphatase (ALP) activities were not influenced by the dietary protein, but proteolytic and acid phosphatase (ACP) activities varied (P<0.05) between the treatments. Proteolytic activity showed a second order polynomial relationship with dietary crude protein (CP) as Y = 0.0734X(2) + 4.937X - 68.37, r(2)=0.97. A positive correlation was observed between dietary CP and amylase (r(2)=0.78). All the metabolites except muscle glucose showed significant change corresponding to the dietary protein levels. Glucose and glycogen levels corresponded to the dietary carbohydrate levels. Muscle and plasma pyruvic acid increased as the crude protein in the diet increased, whereas liver pyruvic acid showed the opposite trend. Muscle protein content was not affected by dietary CP. Protein fractions in plasma (total protein, albumin and globulin) showed maximum values in 30% CP fed group. It is concluded that proteolytic activity and ACP are the major digestive enzymes responsive to dietary CP in L. rohita fingerlings. Considering the cost effectiveness of the diet, and based on liver and plasma free amino acid levels and plasma protein fractions, 30% crude protein is recommended as the optimal dietary protein for L. rohita fingerlings.     

Changes in amino acid composition in the tissues of African catfish (Clarias gariepinus) as a consequence of dietary L-carnitine supplements

A study was undertaken to examine the effect of different amounts of dietary lysine (13 and 21 g kg^?1 diet), lipid (80 and 160 g kg^?1 diet) and L ‐carnitine (0.2 and 1.0 g kg^?1 diet) on growth performance, proximate composition and amino acid metabolism of the African catfish (Clarias gariepinus). Juvenile African catfish (23 ± 1.5 g/fish) were stocked into 70‐L aquaria (16 aquaria, 28 fish/aquarium) connected to a recirculation system during a maximum period of 74 days. All groups were fed at a level of 24 g kg^?0.8 day^?1 in an experiment run at pair feeding. Animals receiving 1.0 g carnitine accumulated up to six times more carnitine in their tissues than animals receiving 0.2 g (P < 0.05). Acyl‐carnitine and free L ‐carnitine levels increased in the whole body and in tissues. Dietary L ‐carnitine supplements increased protein‐to‐fat ratios in the body, but did not affect growth rate. Protein‐to‐fat ratios were only affected when the biosynthesis capacity of L ‐carnitine was restricted due to low lysine levels and when there was a shortage of dietary fat. When lysine was offered at 21 g kg^?1 feed, dietary L ‐carnitine supplements did not affect the amino acid concentrations of body tissues. Dietary L ‐carnitine supplements raised the concentration of glutamic acid > aspartic acid > glycine > alanine > arginine > serine > threonine in skeletal muscle tissue (P < 0.05). Total amino acid concentration in muscle and liver tissues (dry‐matter basis) increased from 506 to 564 and from 138 to 166 mg g^?1, respectively, when diets were offered with high L ‐carnitine, low lysine and low fat levels. These data suggest that dietary L ‐carnitine supplementation may increase fatty acid oxidation and possibly decrease amino acid combustion for energy.     

Effects of dietary taurine on tissue taurine and free amino acid levels of the chum salmon, Oncorhynchus keta, reared in freshwater and seawater environments

1. Young chum salmon were fed on the basal and taurine-supplemented diets for 30 days in freshwater (FW) and for 25 days in seawater (SW). Levels of taurine, major free amino acids (FAA) and non-protein nitrogen (NPN) in various tissues were determined. 2. Tissue taurine levels were higher when fish were fed on the taurine-supplemented diets. All tissues of the SW fish did not contain higher taurine levels than those of the FW. 3. Levels of major FAA in the tissues differed little between fish fed on the basal and taurine-supplemented diets and also between the FW and SW fish. 4. No difference was observed in tissue NPN levels between fish ingesting the basal and taurine-supplemented diets; the levels were slightly higher in the SW fish.     

Digestive enzymes and metabolic profile of Pseudoplatystoma corruscans (Teleostei: Siluriformes) in response to diet composition

Digestive enzyme responsiveness to feeding and associated adjustments of metabolism can be used to derive nutritionally effective diet formulations. Juvenile pintado (Pseudoplatystoma corruscans) were fed different diets. After feeding, fish were killed and blood, liver and white muscle were collected to evaluate metabolites. Stomach along with anterior, middle and posterior intestine were sampled for enzyme analysis. Non-specific protease, trypsin, chymotrypsin, amylase and lipase were assayed. Crude protein (CP) did not induce proteolytic activity; highest protease activities were observed in the stomach. Amylase was higher in the stomach in fish feeding on diets containing 13-25% starch. Lipase activity was observed along the gastrointestinal tract, with the highest activities observed in the middle section. The metabolic profile of white muscle was not affected by CP. In contrast, some plasma and liver metabolites were altered concomitant with changes in the digestive enzymes. Amino acid catabolism was increased. Digestion in pintado was responsive to cornstarch, reflected in intermediary metabolism; proteolytic activities of the digestive tract seem to be sufficient to deal with large amounts of dietary protein. As a result, we are able to recommend a balance between protein and energetic compounds, such as lipids and carbohydrates, in the diet to optimize fish growth.     

Dietary protein reduction in sheep and goats: different effects on <Emphasis Type="SmallCaps">l</Emphasis>-alanine and <Emphasis Type="SmallCaps">l</Emphasis>-leucine transport across the brush-border membrane of jejunal enterocytes

It was the aim of this study to examine the potential regulatory effects of a long-term low dietary protein supply on the transport capacity of the jejunal brush-border membrane for amino acids. For this purpose, we used the neutral amino acids L-alanine (representative for nonessential amino acids) and L-leucine (representative for essential amino acids) as model substances. Ten sheep lambs, 8 weeks of age and 19-27 kg body weight, were allotted to two dietary regimes with either adequate or reduced protein supply which was achieved by 17.9% and 9.7% of crude protein in the concentrated feed, respectively. The feeding periods were 4-6 weeks in length. Similarly, eight goat kids of 5-7 weeks of age and 8-14 kg body weight were allotted to either adequate (crude protein 20.1%, feeding period 9-12 weeks) or reduced protein supply (10.1%, feeding period 17-18 weeks). Dietary protein reduction in lambs caused a significant body weight loss of 0.6 +/- 0.7 kg, whereas the body weight in control animals increased by 1.9 +/- 0.7 kg (P<0.05). Plasma urea concentrations decreased significantly by 60% (low protein 2.3 +/- 0.1 versus control 5.7 +/- 0.2 mmol l(-1), P<0.001). In kids, reduction of dietary protein intake led to significant decreases of the daily weight gain by 48% from 181 +/- 8 g to 94 +/- 3 g (P<0.001) and daily dry matter intake by 27% from 568 +/- 13 g to 417 +/- 6 g (P<0.01). Respective urea concentrations in plasma were reduced by 77% from 5.2 +/- 0.4 to 1.2 +/- 0.2 mmol l(-1) (P<0.01). Kinetic analyses of the initial rates of alanine uptake into isolated jejunal brush-border membrane vesicles from sheep and goats as affected by low dietary protein supply yielded that the apparent Km was neither significantly different between the species nor significantly affected by the feeding regime thus ranging between 0.12 and 0.16 mmol.l(-1). Reduction of dietary protein, however, resulted in significantly decreased Vmax values of the transport system by 25-30%, irrespective of the species. Kinetic analyses of the initial rates of leucine uptake into jejunal brush-border membrane vesicles from sheep and goats yielded that leucine uptake was mediated by Na+-dependent as well as Na+-independent processes. Similar to alanine, apparent Km values of leucine uptake were neither different between the species nor affected due to low dietary protein and ranged between 0.08 and 0.15 mmol l(-1). In contrast to the alanine transport mechanism, dietary protein reduction resulted in increased Vmax values of Na+-dependent leucine transport by 53% in sheep and 230% in goats. Similarly, Na+-independent leucine uptake was stimulated by 85% and 200% in sheep and in goats, respectively. This study shows adaptation of amino acid absorption at the brush-border membrane level of jejunal enterocytes of small ruminants due to dietary protein reduction. Whereas the transport capacity for the nonessential amino acid alanine was reduced due to low dietary protein, the transport capacity for the essential amino acid leucine was markedly stimulated. From this, the involvement of rather different feedback mechanisms in adaptation of intestinal amino acid transport mechanisms has to be discussed.     

Growth depression in socially subordinate rainbow trout Oncorhynchus mykiss: more than a fasting effect

To assess the effects of subordinate social status on digestive function, metabolism, and enzyme activity in salmonid fish, juvenile rainbow trout Oncorhynchus mykiss were paired with size-matched conspecifics (<1.5% difference in fork length) for 5 d. Fish that were fasted for 5 d and fish sampled directly from the holding tank were used as control groups. Both subordinate and fasted fish experienced significant decreases in intestine mass (P = 0.043), and the gall bladder showed marked and significant changes in both size (P = 0.004) and appearance. These findings suggest that the negative effect of social subordination on digestive function reflects in large part a lack of feeding. Hepatic phosphoenolpyruvate carboxykinase activity was significantly higher in subordinate fish relative to dominants, whereas subordinate hepatic pyruvate kinase activity was significantly lower; activities of both enzymes were significantly correlated with plasma cortisol concentrations and behavior scores. Dominant-subordinate differences in the activities of these enzymes were eliminated by administration of the glucocorticoid receptor antagonist RU486, underlining a role for circulating cortisol in eliciting the differences. Significant increases relative to control fish were also detected in red and white muscles from subordinate fish in the activities of protein catabolic enzymes (aspartate aminotransferase, alanine aminotransferase, glutamate dehydrogenase). These differences occurred in the absence of any change in plasma free amino acid or ammonia concentrations, supporting an enhanced turnover of amino acids in muscle in subordinate fish. The results support the hypothesis that changes in metabolism, beyond those elicited by low food consumption, may be responsible at least in part for the low growth rates typical of subordinate fish and that these changes may be related specifically to circulating cortisol levels in subordinate fish.     

Juvenile growth and aggression in diploid and triploid Chinook salmon Oncorhynchus tshawytscha (Walbaum)

Multilocus heterozygosity, aggressive and feeding behaviour, plasma cortisol levels and growth rate were evaluated among three groups of juvenile Chinook salmon Oncorhynchus tshawytscha : diploid, triploid and mixed groups of diploid and triploid fish. There was no difference between diploid and triploid fish in measurements of heterozygosity calculated using seven microsatellite loci, and these measurements did not correlate with performance measurements including feeding rate and growth rate. Aggression trials that examined small groups of fish revealed that after 4 days together in tanks, triploid fish were significantly less aggressive during feeding than diploid fish or fish in mixed groups. At the end of the trials, however, plasma cortisol levels did not differ among the three groups. Thirty-day growth trials in duplicate tanks of 60 fish revealed no difference in growth rate among diploid, triploid and mixed groups, but plasma cortisol levels were significantly lower in triploid fish than in either diploid fish or the mixed fish. Overall, independent of the above differences in aggressive behaviour and cortisol levels, these results suggest similar performance in diploid and triploid Chinook salmon, and thus provide support for the viability of triploid Chinook salmon culture in commercial aquaculture.