The dietary requirement of young rainbow trout (Oncorhynchus mykiss) for folic acid.

We sought to determine the dietary folic acid requirement of young rainbow trout using growth indices supported by measurements of tissue folate concentrations. The investigation was conducted with purified diets that had, by assay, basal folic acid levels of 0.08 and 0.16 mg/kg in the first and second, respectively, of two experiments. Each experiment was started with fry (initial mean weight, 1.4 and 2.8 g/fish in Experiments 1 and 2, respectively) and was conducted at a water temperature of 15 degrees C. Experiment 1 lasted 18 wk and Experiment 2 lasted 16 wk. Recovery tests (of 8 wk duration, performed on fish fed the unsupplemented diet) and pair-feeding showed that the unsupplemented diet led to a folate-specific deficiency condition in which the main hematological abnormality was the appearance of misshapen nuclei in a small proportion (2.3%) of erythrocytes. Dietary requirements were shown not to exceed 0.3 and 0.6 mg folic acid/kg (17 and 33 micrograms/MJ digestible energy) for optimizing survival and growth indices, respectively. We conclude that the dietary folate requirement of the trout is comparable to that of other vertebrates for the purpose of achieving maximal weight gain.

Capacity of the European eel (Anguilla anguilla) to elongate and desaturate dietary linoleic acid.

The ability of eels (Anguilla anguilla) to further desaturate and chain elongate linoleic acid, 18:2n-6, was studied by feeding diets containing either corn oil or a fish oil to groups of elvers for 12 wk and analyzing proportions of fatty acids in tissue lipids. Over the 12-wk period elvers given both dietary treatments increased in weight by fourfold. The proportion of arachidonic acid, 20:4n-6, present in polar lipids of elvers fed the diet containing corn oil increased from an initial value of 5% by weight to 12% by weight; the corresponding value for elvers given a diet containing fish oil, with little linoleic acid in it, was less than 4% by weight. The capacity of this fish to modify dietary linoleic acid metabolically was confirmed by examining the metabolic fate of radioactive carbon after giving [1-14C]linoleic acid orally to eels. Seven days after administration of the linoleic acid approximately 10% of the radioactivity recovered in liver fatty acids was present in trienes and tetraenes with about 4% occurring in arachidonic acid. Compositional analyses from the feeding experiment also indicated that dietary docosaenoic acid, 22:1n-11, is preferentially oxidized by eels.

Effect of selenium deficiency on hydroperoxide-stimulated release of glutathione from isolated perfused liver of rainbow trout (Salmo gairdneri).

1. Duplicate groups of rainbow trout (Salmo gairdneri) were each given partially purified diets which were either adequate or depleted in selenium for 40 weeks. 2. Although there was no significant difference in weight gain, liver Se concentration was significantly lower in fish given the deficient diet. 3. Glutathione (GSH) peroxidase (EC 1.11.1.9) activity was significantly reduced in liver of Se-deficient fish but a differential assay did not indicate the presence of a non-Se-dependent GSH peroxidase activity, although liver GSH S-transferase (EC 2.5.1.18) was significantly increased. 4. Perfusion of livers from trout given Se-adequate diets with t-butyl hydroperoxide (BuOOH) or hydrogen peroxide caused an increase in the rate of release of glutathione disulphide (GSSG) into the perfusate. 5. Perfusion of livers from Se-deficient trout with BuOOH or H2O2 did not result in any change in rate of release of GSSG into the perfusate. 6. These findings confirm the absence of any compensatory non-Se-dependent peroxidase activity in Se-depleted trout.

Some effects of selenium deficiency on glutathione peroxidase (EC 1.11.1.9) activity and tissue pathology in rainbow trout (Salmo gairdneri).

1. Two duplicate groups of rainbow trout (Salmo gairdneri; mean weight 27 g) were given diets of differing selenium content (deficient 0.025 mg Se/kg; supplemented 1.022 mg Se/kg) for 30 weeks. 2. There were no significant differences between treatments in weight gain but packed cell volume, liver vitamin E and liver and plasma Se concentrations were all significantly lower in the Se-deficient trout. 3. Ataxia occurred in about 10% of the Se-deficient trout and histopathologies were evident in nerve cord (damage to axon sheath) and liver (loss of integrity in endoplasmic reticulum and mitochondria with appearance of increased vesiculation). 4. Glutathione peroxidase (EC 1.11.1.9) activity was significantly reduced in liver and plasma of Se-deficient fish but there was no indication, from differential assay, of any non-Se-dependent glutathione peroxidase activity. Glutathione transferase (EC 2.5.1.18) activity was significantly increased in Se-deficient trout.

Some effects of vitamin E and selenium deprivation on tissue enzyme levels and indices of tissue peroxidation in rainbow trout (Salmo gairdneri).

Duplicate groups of rainbow trout (Salmo gairdneri) (mean weight 11 g) were given for 40 weeks one of four partially purified diets that were either adequate or low in selenium or vitamin E or both. Weight gains of trout given the dually deficient diet were significantly lower than those of trout given a complete diet or a diet deficient in Se. No mortalities occurred and the only pathology seen was exudative diathesis in the dually deficient trout. There was significant interaction between the two nutrients both with respect to packed cell volume and to malondialdehyde formation in the in vitro NADPH-dependent microsomal lipid peroxidation system. Tissue levels of vitamin E and Se decreased to very low levels in trout given diets lacking these nutrients. For plasma there was a significant effect of dietary vitamin E on Se concentration. Glutathione (GSH) peroxidase (EC 1.11.1.9) activity in liver and plasma was significantly lower in trout receiving low dietary Se but was independent of vitamin E intake. The ratios of hepatic GSH peroxidase activity measured with cumene hydroperoxide and hydrogen peroxide were the same for all treatments. This confirms the absence of a Se-independent GSH peroxidase activity in trout liver. Se deficiency did not lead to any compensatory increase in hepatic GSH transferase (EC 2.5.1.18) activity; values were essentially the same in all treatments. Plasma pyruvate kinase (EC 2.7.1.40) activity increased significantly in the trout deficient in both nutrients. This was thought to be due to leakage of the enzyme from the muscle and may be indicative of incipient (subclinical) muscle damage.

The effect of vitamin E and oxidized fish oil on the nutrition of rainbow trout (Salmo gairdneri) grown at natural, varying water temperatures.

Groups of rainbow trout (Salmo gairdneri) of approximate mean initial weight 8 g were grown in outdoor tanks over a 14-week period at water temperatures between 12 degrees (start) and 6 degrees (end). Four diets were used. Two contained non-oxidized fish oil (120 g/kg) with or without supplementary DL-alpha tocopheryl acetate and two contained moderately oxidized fish oil again with or without DL-alpha-tocopheryl acetate. The measured selenium content of the diets was 0.10 mg/kg. No significant differences occurred as a consequence of the use of moderately oxidized oil compared with the corresponding treatments using non-oxidized oil. Significant differences did occur between dietary treatments that contained supplementary DL-alpha-tocopheryl acetate and those that did not. These differences applied to weight gain, haematocrit, erythrocyte fragility, mortalities, liver and muscle tocopherol concentrations and lipid peroxidation of liver mitochondria in vitro. Liver glutathione peroxidase (EC 1.11.1.9) activity was unaffected by the dietary treatments used and the proportions of fatty acids in polar lipids of liver and muscle were little changed by the diets used. Severe muscle damage occurred in trout given diets lacking supplementary DL-alpha-tocopheryl acetate. Previous experiments carried out on rainbow trout at a constant water temperature of 15 degrees ( Hung et al. 1981; Cowey et al. 1981, 1983), using diets lacking supplementary vitamin E, did not lead to differences in weight gain, pathological changes or mortalities. Vitamin E requirement may increase as water temperature decreases; minimum dietary requirements for vitamin E measured at a constant water temperature of 15 degrees may not be valid under practical conditions where water temperatures vary over the year.

Tissue distribution, uptake, and requirement for alpha-tocopherol of rainbow trout (Salmo gairdneri) fed diets with a minimal content of unsaturated fatty acids.

The metabolism of and requirements for alpha-tocopherol in rainbow trout fed diets containing 1% linolenic acid as sole source of unsaturated fat and graded levels of tocopherol (0.06-10 mg/100 g) were examined. Fish grew 5-fold over a 16-week period. In liver, tocopherol was concentrated in mitochondria with little in cytosol. Orally administered [3H]-tocopherol was rapidly taken up by plasma and liver but uptake into erythrocytes and white muscle was much slower; in most tissues radioactivity reached a plateau after about 3 days but in red muscle radioactivity increased over a 10-day period. Activities of enzymes that prevent free radical initiated tissue damage did not change in tocopherol deficiency. Tocopherol-deficient trout had no gross or subcellular pathologies even though liver and muscle were severely depleted of the vitamin. Ascorbic acid-stimulated lipid peroxidation in liver organelles indicated a tocopherol requirement of 2-3 mg/100 g diet; the molar ratios of polyunsaturated fatty acids to tocopherol in livers of trout fed diets lacking or supplemented with tocopherol (100 mg/100 g) were 980 and 170, respectively.

Studies on the nutrition of salmonid fish. The magnesium requirement of rainbow trout (Salmo gairdneri).

1. Rainbow trout (Salmo gairdneri) of mean initial weight 35 g were given one of five experimental diets for 20 weeks. The diets contained (g/kg dry diet) 15 calcium, 10 phosphorus and graded levels of magnesium from 0.04 (diet no. 1) to 1.0 (diet no. 5). In a second experiment rainbow trout of mean initial weight 16 g were given one of six experimental diets for 20 weeks. The diets contained (g/kg dry diet): Ca (40), P (30) and levels of Mg from 0.06 (diet no. 6) to 2.0 (diet no. 11). 2. In both experiments weight gains were lowest in those trout given diets containing the basal levels of Mg (diet no. 1 and diet no. 6) but increased with increasing dietary Mg concentration. In neither experiment was there any further increase in weight gain once the Mg concentration reached 0.25-0.5 g/kg dry diet; weight gain reached a plateau at this dietary MG level. 3. The following trends occurred in serum electrolyte concentrations as dietary Mg increased. Mg increased in both experiments, in Expt 2 it reached a maximum of 1 mmol/l when the diet contained 0.5 g Mg/kg and did not increase further; sodium was positively correlated in both experiments; potassium decreased and in Expt 2 reached a plateau minimum of 1.7 mmol/l at a dietary Mg concentration of 0.5 g/kg; Ca and P altered little in either experiment. 4. In both experiments renal Ca concentrations were greatly increased in trout given diets lacking supplementary Mg; they fell to low levels (3-5 mmol/kg) when diets contained 0.15 g Mg/kg or more. Renal K and P concentrations were negatively correlated with dietary Mg in Expt 2; other electrolytes measured were not altered in concentration by the treatments used. 5. Extracellular fluid volume (ECFV) of muscle was negatively correlated with dietary Mg. In Expt 2 it reached a minimal or normal value at 0.5 g Mg/kg diet and did not decrease further. Muscle Mg concentration increased with diet Mg in both experiments and muscle K concentration was also correlated with diet Mg in Expt 2. These changes were related to the shift in muscle water. In Expt 1, P concentration was decreased with increasing diet Mg but in Expt 2 its concentration increased, these changes may have been connected with the three-fold difference in dietary P in the two experiments. 6. By contrast with skeletal muscle, Mg levels in cardiac muscle increased at low dietary Mg intakes. 7. Concentrations of electrolytes in liver did not alter with the dietary treatments used. 8. The results show that Mg requirement of rainbow trout is met by a diet containing 0.5 gMg/kg diet.