Some properties of the intestinal proteases of the rabbitfish,Siganus canaliculatus (Park)

Some properties of the intestinal proteases of the rabbitfish were examined. At 25°C, both trypsin and chymotrypsin showed pH optima of 8.0. Leucine aminopeptidase, however, displayed maximum activity in the pH range, 7.0–9.0. Leucine aminopeptidase had the highest optimum temperature (60°C), and chymotrypsin, the lowest (30°C). The optimum temperature of trypsin was 55°C. The activation energy, E_a, was found to be 8.24 for trypsin and 8.50 kcal mol^–1 for chymotrypsin. The E_a for leucine aminopeptidase was 6.29 kcal mol^–1 above 40°C and 1.73 kcal mol^–1 below 40°C. Substrate concentration-velocity plots showed that all three enzymes followed Michaelis-Menten kinetics; the K_m and V_max were estimated for the three enzymes. The effects of various protease inhibitors on enzyme activity were also examined and confirmed the protease classes to which each enzyme belonged. The three proteases examined have similar properties to proteases in other fishes.     

Inflexibility of vertebral number in chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>

Chum salmon Oncorhynchus keta eggs from the Chitose and Shiriuchi rivers in Hokkaido, northern Japan, were incubated under identical conditions with the aim of comparing vertebral number (V _N) to that reported archived data from the 1950s. The comparison revealed that mean V _N of salmon from the Shiriuchi River has remained unchanged for 50 years and is higher than that from the Chitose River. The V _N in the Chitose River salmon was found to be significantly higher than that recorded in the archival data. The variation in the mean V _N of chum salmon fry collected during different seasons and years in the Chitose River falls within a range that has remained constant, suggesting that the difference between the values obtained in our study and the archival data may be due to differences in the timing of egg collection. These results indicate that the mean V _N of chum salmon has been stable for a long period. Incubation experiments under identical conditions are a useful tool for examining changes in genetic structure and detecting the essential properties of populations.     

Hepatic 5′-monodeiodinase activity in teleosts in vitro: A survey of thirty-three species

The in vitro hepatic 5′-monodeiodination of thyroxine (T₄) to triiodothyronine (T₃) in Oreochromis mossambicus, Channa striata, Clarias batrachus, Cyprinus carpio and Oxyeleotris marmorata was found to be time, pH and temperature dependent, and related to the amount of substrate (T₄) and homogenate introduced into the reaction vessel, in a manner which was consistent with Menton-Michaelis kinetics, and thus indicative of an enzyme-regulated process. Dithiothreitol introduced into the reaction vessel stimulated T₃ production in a dose-related manner. Hepatic 5′-monodeiodinase activity was also detected in a further 28 species of teleosts suggesting that the peripheral monodeiodination of T₄, which is well-documented in salmonids, is also widespread amongst other teleost fishes. All species examined exhibited evidence of enzymatic deiodination, but there were marked differences in K_m and V_max values between the species. There was no apparent phylogenetic or environmental relationships to explain the widely divergent K_m and/or V_max values, nor was there a correlation between K_m and V_max when the species were considered together.     

Sublethal effects of ammonia in soft water on Atlantic salmon smolts at a low temperature

Atlantic salmon (Salmo salar L.) smolts were exposed to ammonia at 4 °C in an open flow system. The concentrations of un-ionized ammonia nitrogen (NH₃-N) and total ammonia nitrogen (TAN) were increased abruptly twice during a period of 28 days. For concentrations in the range 1–15 µgl^–1 un-ionized ammonia nitrogen (4.3–21.1 mgl^–1 TAN), no effects were observed on the smolts. However, plasma glucose was not measured in this range. The observed results for higher concentrations may be divided into two categories of sublethal effects. (1) In smolts exposed to the concentration range 19–37 µgl^–1 NH₃-N (average 26 µgl^–1 NH₃-N; 34.9–50.5 mgl^–1 TAN), the mean plasma glucose concentration was 1.3 times the mean of the control group. No other measured parameters were significantly altered. (2) For smolts exposed to 37–65 µgl^–1 NH₃-N (average 47 µgl^–1 NH₃-N; 57.7–84.5 mgl^–1 TAN), the mean plasma glucose concentration was increased to 2.3 times, the mean plasma chloride level was reduced to 0.9 times, and mean haematocrit was increased to 1.1 times the control values. Histological examination of the gill tissue revealed extensive lamellar hypertrophy. Weight, length, condition factor and ventilation frequency were not significantly affected and no mortality took place during the exposure period (1–65 µgl^–1 NH₃-N; 4.3–84.5 mgl^–1 TAN). Plasma glucose seemed to be the most sensitive parameter in detecting the sublethal stress response, and this investigation may thus demonstrate the importance of measuring plasma glucose in experiments concerning safe levels for ammonia.     

The cytochrome P450 system of atlantic salmon (Salmo salar): I. Basal properties and induction of P450 1A1 in liver of immature and mature fish

The major components of the cytochrome P450 (P450) system in liver microsomes of Atlantic salmon were studied using spectrophotometric, catalytic and immunochemical techniques. In juvenile fish sampled during the winter season, high basal activities of 7-ethoxyresorufin O-deethylase (EROD) were found. The K_m for 7-ethoxyresorufin was 0.4 µM, and V_max 1.23 nmol/min/mg protein in juvenile fish. In mature fish sampled from the same group of fish in December, EROD activity was barely detectable (20–30 pmol/min/mg protein). Treatment with the P450 1A1 inducer -naphthoflavone (BNF) resulted in almost 2-fold induction of total P450, and 30–40-fold induction of EROD activity in immature fish. A similar fold increase was seen in mature fish. The differences in EROD activity between untreated and BNF-treated fish, was accompanied by similar differences in a P450 1A1 cross-reacting protein (M_r=58,000 D) in immunochemical studies using rabbit anti-cod P450 1A1 IgG. However, judging from these studies, the levels of P450 1A1-protein in mature salmon far exceeded those accounted for by the measured EROD activity in comparison to immature fish (both before and after BNF-treatment), indicating inhibiting effects of sex steroids on the measured activity. This effect was not seen on 7-ethoxycoumarin O-deethylase activity. A long-term storage experiment indicated that Atlantic salmon liver microsomes can be stored for 2 years at –80°C in 20% glycerol without losing more than 20–40% of its catalytic activity.Parts of this work were presented at the 5th International Symposium on Responses of Marine Organisms to Pollutants, April 1989 in Plymouth, United Kingdom (Larsen and Goksøyr 1989).     

Protein content and freezing avoidance properties of the subdermal extracellular matrix and serum of the Antarctic snailfish,Paraliparis devriesi

The Antarctic snailfish, Paraliparis devriesi (Liparididae), occupies an epibenthic habitat at a depth of 500–650 m in the subzero waters of McMurdo Sound, Antarctica. This species has watery (97%) gelatinous subdermal extracellular matrix (SECM) comprising a mean of 33.8% of the body weight, the largest known proportion of any adult fish. The protein concentration of the SECM was found to be 6–7 mg ml^–1 (0.6–0.7% w/v). Separation of the polypeptides of the SECM by SDS-PAGE revealed 11 polypeptides ranging in relative molecular mass (M_r) from 67,000 to 13,000, with other unresolved polypeptides of less than 13,000. The isoelectric points of these proteins ranged from 4.85 to 8.05. Partial N-terminal amino acid sequence data were obtained for four of the major SECM polypeptides. The N-terminal amino acid sequences of three of these were not identical to or homologous with any other known sequences, whereas the N-terminal sequence of one polypeptide (M_r 51,000) was identical to partial sequence from the apolipoprotein A-I precursor of Atlantic salmon (Salmo salar). Although not isolated from either SECM or serum, melting point-freezing point behavior of body fluids suggest that Paraliparis possess modest amounts of a noncolligative antifreeze compound. Since relatively small amounts of antifreeze are present in the serum and even less in the SECM, freezing avoidance results from the combined effects of antifreeze and the elevated osmolality of body fluids. There are no special adaptations to prevent freezing in the superficially located high water content SECM.     

Metabolic biochemistry of cardiac muscle in three tuna species (bigeye, Thunnus obesus ; yellowfin, T. albacares ; and skipjack, Katsuwonus pelamis ) with divergent ambient temperature and oxygen tolerances

Bigeye tuna (Thunnus obesus) have much greater vertical mobility than yellowfin (T. albacares) and skipjack (Katsuwonus pelamis) tunas, due to an apparent greater tolerance of the changes in ambient temperature and oxygen occurring with depth. In an attempt to identify physiological processes (e.g., effects of temperature on cardiac function) responsible for these behavioral differences, we examined enzyme activities (at 12 °C, 17 °C, and 25 °C) of cardiac muscle in all three species. Contrary to our expectations, we found few differences and no clear explanatory patterns in maximum enzyme activities (V_max) or enzyme activity ratios. For example, citrate synthase (CS) activity was the same in bigeye and skipjack tunas, but 40% lower in yellowfin tuna, whereas carnitine palmotoyltransferase (CPT) activity in skipjack tuna was approximately double that in the other two species. The ratio of CPT to pyruvate kinase (PK) activity, a measure of the tissues preference for fatty acids as metabolic substrates, was the same in bigeye and yellowfin tunas, but elevated skipjack tuna. The ratios of lactate dehydrogenase (LDH) to CS activity and of PK to CS activity (anaerobic–aerobic enzyme activity ratios – taken as measures of the tissues ability to tolerate hypoxia) were both elevated in yellowfin tuna cardiac tissue relative to the other two species. We also found no differences in temperature sensitivity (Q₁₀ values) when comparing cardiac enzyme activities across species, nor effects of temperature on the substrate affinity (K_m) of LDH. In sum, our results do not suggest any clear metabolic difference in the cardiac muscle that would explain the apparent greater tolerance of bigeye tuna to acute hypoxia and ambient temperature changes or their substantially greater vertical mobility.     

Seasonal variations in and effect of incubation water temperature on vertebral number in naturally spawning chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>

Seasonal variations and reaction norms for vertebral number (V _N) in response to incubation water temperature were estimated in adult and juvenile naturally spawning chum salmon Oncorhynchus keta. The mean V _N of adults varied according to spawning time; the early-spawning population had higher V _N values than the late-spawning population. Moreover, the mean V _N values in the early-spawning population decreased with seasonal changes, whereas V _N values in the late-spawning population remained stable. Chum salmon embryos in three full-sib families were incubated at five different temperatures until hatching, and the V _N values of the resulting juveniles were analyzed. The V _N reaction norm to incubation water temperature showed a V-shaped curve that was lowest at an intermediate temperature. The mean V _N at the same incubation temperature varied among the three families. These results suggest that V _N values in chum salmon are influenced by genetic components and incubation water temperatures. V _N may be a useful parameter for estimating the environmental conditions during ontogenesis and the genetic background by detecting population changes.     

Adenosine triphosphate levels of chinook salmon (Oncorhynchus tshawytscha) eggs following in vitro maintenance and activation/fertilization

To further examine the concept of egg quality and the physiology of stored salmonid eggs, we investigated the effects of different oxygen tensions on the adenosine triphosphate (ATP) levels of unfertilized, activated, and fertilized chinook salmon (Oncorhynchus tshawytscha) eggs. The ATP levels of unfertilized chinook salmon eggs were 2.61±0.14 nmol ATP per egg (17.6±0.9 mol l^–1 relative to cell water) and ranged from 1.98 to 3.63 nmol ATP per egg. The ATP content of unfertilized eggs maintained at 10 °C under 100% O₂, 21% O₂, and 100% N₂ remained unaltered throughout a 120 h storage period. Storing eggs under identical conditions at 20 °C (in an effort to speed egg metabolism and ATP turnover) resulted in significant O₂-independent decreases in ATP levels. However, ATP levels of unfertilized eggs exposed to 1 mmol l^–1 potassium cyanide (a potent inhibitor of oxidative phosphorylation) at 10 °C were significantly decreased after 24 h and continued to decline throughout the 120 h maintenance period to about 30% of time=0 values. Maintenance with exogenous nutrients (5 mmol l^–1 acetate plus 5 mmol l^–1 pyruvate) over 120 h at 10 °C did not alter the ATP content of unfertilized eggs. Eggs activated by exposing them to 10 °C water for a few minutes showed a rapid decrease in ATP values, regardless of whether the eggs were fertilized or not. Following an initial 25% drop after fertilization, the ATP levels remained stable for the remainder (5 d) of the incubation period in eggs maintained in 10 °C water. Therefore, unfertilized chinook salmon egg ATP levels appear to be relatively stable and maintained by a low, cyanide-inhibitable metabolism. The stability of egg ATP levels may be one reason that salmonid eggs can be stored for several days while eggs from other fishes cannot.     

Effects of acclimation temperatures of 7 and 14 °C on the contractility of Atlantic salmon (Salmo salar) intestinal smooth muscle in vitro

The contractility of intestinal smooth muscle of Atlantic salmon (Salmo salar) acclimatized to temperatures of 7 and 14 °C was compared over a prolonged period of time (9 months) in freshwater. Initial studies were carried out on intestine isolated from Atlantic salmon to estimate optimal conditions for contractility. Subsequent studies were carried out with the neurotransmitter/agonist 5-HT and TS, which activates neuronal elements in the intestine, in a Krebs-Henseleit solution containing HEPES buffer maintained at pH 7.85 and at the temperature of the experimental group (i.e., 7 or 14 °C). There was a significant interaction with time and temperature for the maximal response of 5-HT (p=0.005), effective concentration producing 50% of the maximal response (EC₅₀) to 5-HT (p=0.026) and maximal response to TS (p=0.002), demonstrating that gastrointestinal contractility of Atlantic salmon is altered by both time (month) and temperature. No significant changes were found with the effective frequency producing 50% of the maximal response (EF₅₀) to TS. The potency (EC₅₀ of 5-HT) increase between 7 and 14 °C was statistically significant (p=0.026) but the small differences found with potency were most likely not physiologically significant. A time shift was recognized in the maximal responses to 5-HT, by applying a time variable to equations for the curves of the responses. This demonstrated that the curves are similar for both temperatures but alterations in the time of the maximal response changes was different. Thus, there is a delay in the time (month) changes of gastrointestinal contractility related to acclimation temperature and would be an area for future study.     

Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Para-nitrophenol hydroxylation by fish liver microsomes: kinetics and effect of selective cytochrome P450 inhibitors

The study investigated the kinetics of p-nitrophenol hydroxylase (PNPH) in hepatic microsomes obtained from Atlantic salmon (Salmo salar). The selective inhibitors for some major mammalian cytochrome P450 (CYP450) were used to investigate the potential inhibitory effect on enzymes involved in p-nitrophenol hydroxylation. The following inhibitors were used: α-naphtoflavone (CYP1A), ellipticine (CYP1A1), furafylline (CYP1A2), 8-methoxypsoralen (8MOP, CYP2A6), 4-methylpyrazole (4MP, CYP2A6/2E1), diallyl sulfide (DAS, CYP2E1), and ketoconazole (CYP3A4). Additionally, the natural steroids 17-beta-oestraiol (E2) and testosterone were investigated as potential inhibitors of PNPH activity. It was found that formation of 4-nitrocatechol from p-nitrophenol followed monophasic kinetics with K _m  = 0.17 ± 0.03 mM and V _max = 21.8 ± 1.05 pmol/min/mg. PNPH activity was competitively inhibited by diallyle sulfide with the K _i value of 285.1 ± 94.2 μM μM and uncompetitively by ellipticine with K _i value of 65.7 ± 7.8 μM. Moreover, E2 showed an ability to reduce PNPH activity through the mechanism-based inhibition mode. Our results suggest that hepatic microsomes from Atlantic salmon possess CYP2E1-like activity. However, specific isoform-mediated PNPH activity should be identified.     

Control of phosphorus homeostasis of Atlantic salmon (Salmo salar) in fresh water

Studies were conducted to determine the absorption, excretion and requirement of dietary phosphorus (P) by Atlantic salmon (Salmo salar). Triplicate groups of salmon parr, initial weight 15 ± 0.5±g , were fed, diets containing 4, 5, 6, 7, 8, 9, 11 and 13 mg P and 20 KJ of digestible energy (DE) per±g of diet (dry matter basis, DMB) to satiation for 16 weeks. The basal diet containing 4 mg P g^-1 (0.15 mg available P per KJ DE) was supplemented with graded levels of calcium phosphate, Ca(H₂PO₄)²H₂O to formulate the eight experimental diets. The fish were reared in fresh water at a temperature of 15 °C on a 12 h photoperiod. Vertebrae ash increased from 316 to 516 mg g^-1 fat-free dry matter with an increase in dietary P content. P requirement was estimated by using a four-parameter sigmoidal equation. The data suggests that a diet of 0.28 mg available P per KJ DE is needed for Atlantic salmon. Phosphate and calcium levels in plasma and bone increased, whereas levels of magnesium and liver cholecalciferol decreased, with an increase in dietary P.Phosphate excretion in urine and apparent availability of P were determined in deficient and replete fish. In deficient fish, the urine phosphate concentration was 0.10 mmol L^-1 before feeding and 0.25 mmol l^-1 after feeding, whereas in replete fish these concentrations were 1.09 and 5.11 mmol l^-1, respectively. The increase in urine phosphate concentration was higher in replete fish than in deficient fish, however, the apparent absorption of P was found to be significantly lower in replete fish than in deficient fish. These results suggest that similarly to terrestrial vertebrates, P homeostasis in Atlantic salmon is controlled by absorption in the intestine, conservation in the kidney and storage in the bones.     

Long-term nutritional effects on the primary liver and kidney metabolism in rainbow trout (Oncorhynchus mykiss). II. Adaptive response of glucose 6-phosphate dehydrogenase activity to high-carbohydrate/low-protein and high-fat/non-carbohydrate diets

The influence of long-term administration of high-carbohydrate/low-protein and high-fat/non-carbohydrate diets were studied in relation to kinetic behaviour of glucose 6-phosphate dehydrogenase in liver and kidney of rainbow trout. In all cases studied, the saturation curves of these enzyme showed typical hyperbolic kinetics without evidence of sigmoidicity. After 30 days of feeding with a high-fat diet (170 g kg^?1), there was a significant decrease in V_max and specific activity (45%) as well as catalytic efficiency (39%) without changes in K_m or activity ratio of hepatic glucose 6-phosphate dehydrogenase. These changes agree more with a clearly decreased cell concentration than with an inhibition of the pre-existing enzyme. The administration of a high-carbohydrate diet (60 g kg^?1), contrary to what was previously thought, decreased V_max by 21% and specific activity and catalytic efficiency by 30%, without significant changes in the other kinetic parameters of the hepatic enzyme. The kinetic behaviour under these nutritional conditons was due to the rejection of this diet by the fish and thus could be considered a low-feeding situation. On the other hand, no variations in the kinetics of renal glucose 6-phosphate dehydrogenase were found, clearly demonstrating that in this organ, the pentosephosphate pathway showed no adaptive response related to fattyacid and other lipid synthesis. The activity of the renal enzyme was consistently half that of the hepatic enzyme.     

Pharmacokinetic model of florfenicol in turbot (Scophthalmus maximus): establishment of optimal dosage and administration in medicated feed

The pharmacokinetics of florfenicol (FF) in turbot (Scophthalmus maximus) was studied after single intravenous (10 mg kg⁻¹) and oral (100 mg kg⁻¹) administration. The plasma concentration–time data of florfenicol were described by an open one‐compartment model. The elimination half‐life (t_1/2) was estimated to be 21.0 h, and the total body clearance, Cl, was determined as 0.028 L kg h⁻¹. The apparent volume distribution (V_d) was calculated to be 0.86 L kg⁻¹ and the mean residence time (MRT_iv) was 30.2 h. Following oral administration, the maximum plasma concentration (C_max) of 55.4 μg mL⁻¹ was reached at 12 h (T_max). The absorption constant (k_a) was 0.158 h⁻¹. The bioavailability was estimated to be 57.1%. The low bioavailability observed at higher doses was explained by the saturation of the mechanisms of absorption. The drug absorption process was limited by its inherent low solubility, which limited the amount of available FF absorbed in the gastrointestinal tract. Based on the pharmacokinetic data, an optimal dosing schedule for FF administration is hereby provided. Based on the minimum inhibitory concentration found for susceptible strains of Aeromonas salmonicida, oral FF administration of first, an initial dose of 30 mg FF kg⁻¹, followed by 6 maintenance doses at 18 mg kg⁻¹/daily could be effective against furunculosis in turbot.     

First multi-generation culture of the tropical cuttlefish Sepia pharaonis Ehrenberg, 1831

Sepiapharaonis, the pharaoh cuttlefish was cultured through multiplegenerations in the laboratory (5 consecutive generations) using closed,recirculating water filtration systems. The eggs of the original parentalgeneration (G_P) were spawned by a wild caught Gulf of Thailandfemale in alocal fisheries laboratory, then packed and shipped air cargo to Texas wherehatching occurred. The culture temperature ranged 25°–28°C, except for one generation that was chilled intentionallyto21 °C and then warmed to 25 °C after 9.6months. Spawning occurred as early as day 161. Spawning output was high in allgenerations except the group that was cultured at 21 °C. Eggfertility was low in captivity (< 20%), but hatchling survival was high(>70%). The average egg incubation time was 13.6 d at 25–28°C. The largest spawn resulted in 600 viable hatchlings andthesmallest resulted in 11 hatchlings. The cuttlefish ate a wide variety ofestuarine crustaceans and fishes as well as frozen shrimp. There were noapparent disease problems since survival from hatching to maturity was over70%.The average life span for cuttlefish cultured at 25–28°Cwas 8.9 months and 12.3 months at 21 °C. Size at hatching wasmeasured for fourth generation (G₄) hatchlings; the mean weight athatching was 0.103 g and the mean mantle length was 6.4mm. The largest cuttlefish cultured was a male 300 mmML and 3,045 g; the oldest cuttlefish lived 340 d.This cuttlefish species presents an excellent choice for commercial mariculturebecause of its rapid growth, short life span, tolerance to crowding andhandling, resistance to disease and feeding habits.     

Glycogenolytic action of glucagon-family peptides and epinephrine on catfish hepatocytes

Glycogenolytic effects of salmon and mammalian glucagons, salmon glucagon-like peptide (GLP) and epinephrine were studied on liver cells isolated from catfish (Ictalurus melas). In spring and summer, salmo-glucagon (3×10^–10 to 3×10^–8 M) was more effective than its mammalian counterpart in the stimulation of glucose release and cAMP synthesis in hepatocytes. GLP was less potent as compared to both glucagons. -amylase activity was not affected by the treatment with either glucagon-family peptides or epinephrine.The comparison of the glycogenolytic effects of salmon glucagon to those of epinephrine reveals a greater potency of the latter hormone in the stimulation of cAMP synthesis, glycogen-phosphorylase activity and glucose release. Glycogen content in the liver cells was equally depleted after treatment with both of the two hormones.     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.     

Effect of Dietary Lipids on Macrophage Function,Stress Susceptibility and Disease Esistance in Atlantic Salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

As the supply of marine fish oil is becoming a limiting factor in the production of Atlantic salmon (Salmo salar), new diets and alternative sources of energy are being tested. Plant oils are natural potential candidates to replace fish oil, but the different levels of essential polyunsaturated fatty acids may influence the health and growth of salmon. In this study, we have investigated the resistance to transport stress and bacterial infection, phagocytic activity in head kidney macrophages and eicosanoid metabolism in salmon fed three different diets. In high-energy fishmeal based diets, 50% and 100% of the supplementary fish oil (FO) was replaced with soybean oil (SO). The three dietary groups were fed for 950 day-degrees at 5 °C (27 weeks) and 12 °C (11 weeks) before challenging the fish with Aeromonas salmonicida, analyzing the lipid composition of head kidney and examining macrophage function in vivo and in vitro. Dietary fatty acids affected the lipid composition of the kidney. The level of eicosanoid precursor’s 20:4n-6 and 20:3n-6 were 3 and 7-fold higher in the 100% SO group compared with the FO group. The total fraction of n-3 lipids in kidney was 19% in the SO group, compared to 16% and 12% in the 50% or 100% SO groups, respectively. However, the production of leucotriene B₄ (LTB) and prostaglandin E₂ (PGE) immunoreactive materiel from exogenously added arachidonic acid in head kidney macrophages was only affected by the composite diet (increased) at 5 °C. In addition, the phagocytic activity of kidney macrophages in vivo and in vitro was not affected by diet. No effect of diet was observed on transport stress or susceptibility to a bacterial infection with Aeromonas salmonicida. Atlantic salmon therefore seems to tolerate a diet solely based on soybean oil as lipid source, without any detrimental effects on growth, health and immune functions.     

Capacity for digestive hydrolysis and amino acid absorption in Atlantic salmon (Salmo salar) fed diets with soybean meal or inulin with or without addition of antibiotics

This experiment was done to study the effects of dietary soybean meal (SBM) and inulin (a prebiotic) on the capacity for digestive hydrolysis and amino acid absorption by Atlantic salmon, and how a dietary supplement of the broad-spectrum antibiotic oxytetracycline (OTC) modulated these responses. A control diet (FM) was made from fish meal, fish oil and extruded wheat. Two similar diets were made with 250 g soybean meal (SBM) or 75 g inulin kg^− 1. Each diet was made with or without a supplement of 3 g OTC kg^− 1. All six diets contained yttrium oxide for estimation of apparent nutrient absorption. Each diet was fed to two groups of 172 g salmon kept in 1 m² tanks with 9 °C saltwater for 3 weeks. Intestinal organs were then sampled and weighed. Gastrointestinal tracts (GIT) were sectioned for analyses of brush border alkaline phosphatase (ALP) and leucine aminopeptidase (LAP) activities. Tissue from the distal intestine (DI) was also fixed for histological examination. Digesta from the different sections were freeze dried for estimation of trypsin and amylase activities, and of apparent absorption of amino acids (AA), nitrogen (N), and sulphur (S). About 85% of the trypsin activity, 70% of the amylase activity, 85% of the ALP activity, and 82% of the LAP activity were found in the proximal (PI) and mid (MI) intestine of fish with functional DI, and the absorption of AA, N, and S was quantitatively completed in the MI. Dietary OTC resulted in lower relative liver weight, but apart from increased ALP and LAP activities in DI when feeding OTC in combination with inulin, OTC did not modify the responses to dietary SBM or inulin. Dietary SBM resulted in lower relative liver weight, and induced pathomorphological changes in the DI mucosa, thus lower the ALP and LAP activities in the DI. SBM also stimulated absorption of AA, N, and S in the PI, but at the same time increased the activities of trypsin and amylase in the DI, indicating reduced re-absorption and increased faecal losses of these endogenous enzymes. Dietary inulin did not damage the DI, and stimulated intestinal growth and higher relative mass of the GIT. Inulin without OTC did not affect the hydrolytic and absorptive capacity of the salmon GIT.