Plasma, liver and kidney folate and plasma homocysteine concentrations are poor response variables at very low dietary folate intakes, in a folate depletion/repletion rat model.

Folate depletion/repletion rat models are popular protocols for assessing the bioavailability of folate. Much of the early work carried out on folate bioavailability concentrated on foods naturally high in folate. However, foods low in folate often contribute significantly to folate intake because of their high consumption in the general population. Therefore, the assessment of the bioavailability of foods low in folate is essential to properly estimate folate intake. The present study investigated plasma, liver and kidney folate and plasma homocysteine concentrations as appropriate response variables for measuring folate bioavailability in the rat at very low dietary folate intakes. One hundred and one weanling male rats (Wistar strain) were fed a folate-deficient diet containing 1% succinyl sulfathiazole for 28 d. Following depletion, six rats were randomly assigned to each of 16 repletion diets containing folic acid, fortified white bread, unfortified wholemeal bread or unfortified rye bread calculated to provide 6.25, 12.5, 18.75 and 25 micrograms folate/kg of each diet. After a further 28 d, plasma, liver and kidney folate concentrations were determined by microbiological assay. Plasma homocysteine was measured by HPLC as a functional indicator of folate status. Only a weak correlation was found between the response variables measured and dietary folate intake, indicating that this folate depletion/repletion rat model is not suitable for testing the response of rats fed diets containing very low levels of folate.

Effects of preparation and cooking of folic acid-fortified foods on the availability of folic acid in a folate depletion/repletion rat model.

The practice of food fortification with folic acid offers the potential to increase the folate intake of the general population. To fully exploit the potential of fortification for raising folate nutriture, appropriate food vehicles need to be selected. Selection should involve determination of the availability of folic acid as affected by characteristics of the carrier food, food matrix, food preparation, and cooking. The present study investigated the effects of preparation and cooking of a range of folic acid-fortified foods on the folate status of folate-deficient rats. Fifty-six weanling male rats (Wistar strain) were fed a folate-deficient diet containing 1% succinyl sulfathiazole for 28 days. Following depletion, six rats were randomly assigned to each of eight repletion diets containing cooked or uncooked meringue mix, quick bread mix, brownie mix, or pizza base mix. The test foods were fortified with 1400 microg of folic acid/kg of food and incorporated as 19% of the repletion diets. Each of the first four groups was pair-fed a diet containing a cooked fortified food with another group fed the corresponding uncooked fortified food. After a further 28 days, plasma, liver, and kidney folate concentrations were determined by microbiological assay. Mean plasma and liver folate concentrations of rats fed diets containing cooked fortified foods were similar to those of rats fed uncooked fortified foods. Preparation and cooking did not affect the availability of folic acid from the selected cereal-based convenience foods in this rat model system, suggesting that these foods are appropriate vehicles for fortification with folic acid.

Influence of folic acid-fortified foods on folate status in a folate depletion-repletion rat model.

An increasing number of foods fortified with varying levels of folic acid are appearing in the market place, targeted either at the general population or at specific consumer groups. Although it is assumed that the folate in these products should be highly bioavailable, there is a need to carry out studies to ascertain that this is, in fact, the case. The present study investigated the ability of selected folic acid-fortified foods (targeted at different types of consumer) to increase the folate status of folate-deficient rats. Forty-two weanling male rats (Wistar strain) were fed a folate-deficient diet containing 1 % succinyl sulfathiazole (w/w) for 28 d. Following depletion, seven rats were randomly assigned to each of five repletion diets containing folic acid, Complan, Slim Fast, Opti-Fuel2 or Cola Coa calculated to provide 200 microg folate/kg of each diet. Calculations were based on folate information from the product labels. After a further 28 d, plasma, liver and kidney folate concentrations were determined by microbiological assay. Plasma homocysteine was measured by HPLC as a functional indicator of folate status. The folate content of the foods was measured by tri-enzyme extraction followed by microbiological assay. Our analyses suggest that there may be considerable inaccuracies on the part of the manufacturers in relation to the folate declarations on the product labels. Despite this, the four foods evaluated were highly effective in elevating plasma, liver and kidney folate and lowering plasma homocysteine concentrations in rats. These results lend support to the policy of food fortification with folic acid as a means of raising the folate status of the population, and in particular to the fortification of specific foods which may target areas of the population where increased folate status is most needed.

Optimal nutrition: vitamin E.

Interest in the role of vitamin E in disease prevention has encouraged the search for reliable indices of vitamin E status. Most studies in human subjects make use of static markers, usually alpha-tocopherol concentrations in plasma or serum. Plasma or serum alpha-tocopherol concentrations of < 11.6, 11.6-16.2, and > 16.2 mumol/l are normally regarded as indicating deficient, low and acceptable vitamin E status respectively, although more recently it has been suggested that the optimal plasma alpha-tocopherol concentration for protection against cardiovascular disease and cancer is > 30 mumol/l at common plasma lipid concentrations in combination with plasma vitamin C concentrations of > 50 mumol/l and > 0.4 mumol beta-carotene/l. Assessment of vitamin E status has also been based on alpha-tocopherol concentrations in erythrocytes, lymphocytes, platelets, lipoproteins, adipose tissue, buccal mucosal cells and LDL, and on alpha-tocopherol: gamma-tocopherol in serum or plasma. Erythrocyte susceptibility to haemolysis or lipid oxidation, breath hydrocarbon exhalation, oxidative resistance of LDL, and alpha-tocopheryl quinone concentrations in cerebrospinal fluid have been used as functional markers of vitamin E status. However, many of these tests tend to be non-specific and poorly standardized. The recognition that vitamin E has important roles in platelet, vascular and immune function in addition to its antioxidant properties may lead to the identification of more specific biomarkers of vitamin E status.

Lipid stability in meat and meat products.

Lipid oxidation is one of the main factors limiting the quality and acceptability of meats and meat products. Oxidative damage to lipids occurs in the living animal because of an imbalance between the production of reactive oxygen species and the animal's defence mechanisms. This may be brought about by a high intake of oxidized lipids or poly-unsaturated fatty acids, or a low intake of nutrients involved in the antioxidant defence system. Damage to lipids may be accentuated in the immediate post-slaughter period and, in particular, during handling, processing, storage and cooking. In recent years, pressure to reduce artificial additive use in foods has led to attempts to increase meat stability by dietary strategies. These include supplementation of animal diets with vitamin E, ascorbic acid, or carotenoids, or withdrawal of trace mineral supplements. Dietary vitamin E supplementation reduces lipid and myoglobin oxidation, and, in certain situations, drip losses in meats. However, vitamin C supplementation appears to have little, if any, beneficial effects on meat stability. The effect of feeding higher levels of carotenoids on meat stability requires further study. Some studies have demonstrated that reducing the iron and copper content of feeds improves meat stability. Post-slaughter carnosine addition may be an effective means of improving lipid stability in processed meats, perhaps in combination with dietary vitamin E supplementation.

Tissue content of alpha-tocopherol and oxidative stability of broilers receiving dietary alpha-tocopheryl acetate supplement for various periods pre-slaughter.

1. The effects of dietary alpha-tocopheryl acetate on the alpha-tocopherol status of chicken plasma and tissues were investigated. The rate of iron-ascorbate-induced lipid peroxidation was also studied. 2. One hundred and forty four chicks were divided into 6 groups: one control group was fed a basal diet of 30 mg alpha-tocopheryl acetate/kg food for the duration of the trial. A supplemental diet of 200 mg alpha-tocopheryl acetate was fed to each of the other 5 groups for 1,2,3,4 or 5 weeks prior to slaughter. 3. Supplementation resulted in an increase in alpha-tocopherol in plasma and all tissues examined. Saturation levels of alpha-tocopherol were observed in plasma after 1 week of feeding and in tissues within 3 to 4 weeks of feeding. 4. Supplementation with alpha-tocopheryl acetate for up to 4 weeks pre-slaughter resulted in significant reductions in susceptibility to induced lipid peroxidation. 5. Overall, the results show that feeding 200 mg alpha-tocopheryl acetate/kg food to chicks for at least 4 weeks prior to slaughter is necessary to optimise muscle content and stability against lipid peroxidation.

Supranutritional vitamin E supplementation in pigs: Influence on subcellular deposition of α-tocopherol and on oxidative stability by conventional and derivative spectrophotometry.

The influence of three levels of vitamin E (30, 200 and 1000mg kg(-1)) in the diet of pigs on the subcellular deposition of α-tocopherol in muscle and on the oxidative stability determined by conventional and first derivative Spectrophotometry was studied. The content of α-tocopherol in m. gluteo biceps and in mitochondrial and microsomal fractions of the muscle significantly increased (p < 0.01) with increasing levels of dietary vitamin E. Concentrations of α-tocopherol in muscle, mitochondria and microsomes of pigs fed diet supplemented with 1000 mg kg(-1) α-tocopheryl acetate were 3.2-, 6.1- and 5.6-fold greater, respectively, than those in their counterparts from the control animals. These differences in α-tocopherol concentration in the subcellular fractions and intact muscle resulted in enhanced stability of the membranes and the tissue when exposed to iron-ascorbate induced peroxidation. When lipid oxidation in the same samples was further measured by the first derivative method, the resultant MDA-TBA values were 59-69% lower in tissue samples, 16-19% lower in mitochondria and 6-9% lower in microsomes than the conventional TBARS values.

Oxidative stability and alpha-tocopherol retention in turkey burgers during refrigerated and frozen storage as influenced by dietary alpha-tocopheryl acetate.

1. The effect of vitamin E (alpha-tocopheryl acetate) in turkey diets on the oxidative stability of raw and cooked turkey burgers and on the retention of alpha-tocopherol during refrigerated (4 degrees C) or frozen (-20 degrees C) storage was investigated. One hundred and two, one-day-old T-8s turkey poults were divided at random into 3 groups of 34 animals each and fed on either a basal diet (normal commercial turkey diet) supplemented with 20 mg alpha-tocopheryl acetate/kg (control) or fed an alpha-tocopherol supplemented diet containing 300 (E300) or 600 (E600) mg alpha-tocopheryl acetate/kg for 21 weeks. 2. Dietary supplementation with alpha-tocopheryl acetate significantly reduced TBARS numbers in both raw and cooked burgers during refrigerated and frozen storage. 3. The mean values of alpha-tocopherol in raw and cooked burgers stored at 4 degrees C did not change during storage. 4. In the case of both raw and cooked samples stored at -20 degrees C, the alpha-tocopherol values decreased from 5.67 to 3.54 and from 3.56 to 2.30 micrograms/g in the raw burgers from turkeys from the E600 and E300 treatments, respectively, after 4 months storage. The values decreased from 5.60 to 2.88 and from 3.29 to 1.85 micrograms/g in cooked burgers from turkeys from the E600 and E300 treatments, respectively, after 5 months storage.

Uptake of α-tocopherol in porcine plasma and tissues.

The effect of feeding α-tocopheryl acetate to pigs on the rate and extent of uptake of α-tocopherol in various tissues was investigated. The rate of iron ascorbate-induced lipid peroxidation was also studied. One hundred and eight Landrace × Large White pigs were assigned at random to one of the following barley-based diets: 20 mg α-tocopheryl acetate/kg feed for up to 126 days; 20 mg/kg feed to day 91, followed by 200 mg/kg feed to day 126, or 200 mg/kg feed for up to 126 days: Pigs from each group were slaughtered at specified intervals. For pigs fed the diet supplemented with 200 mg α-tocopheryl acetate/kg feed, α-tocopherol levels increased with increasing supplementation time up to day 126 in all tissues studied. The highest levels of α-tocopherol were observed in kidney fat and subcutaneous fat (inner layer) followed by subcutaneous fat (outer layer), liver, lung, heart, kidney, with muscle and brain containing approximately the same level. The α-tocopherol concentrations in all tissues examined, from pigs fed 200 mg α-tocopheryl acetate/kg feed from weaning to day 126 were greater than those fed the supplemented diet for 35 days. Iron-induced lipid peroxidation was reduced by dietary α-tocopheryl acetate supplementation, and muscle samples from pigs supplemented for 126 days were significantly (p < 0.05) less susceptible to peroxidation compared to muscle from pigs fed the same supplemented diet for 35 days.

Consumption of thermally-oxidized sunflower oil by chicks reduces alpha-tocopherol status and increases susceptibility of tissues to lipid oxidation.

The effect of heated sunflower oil consumption on alpha-tocopherol status, fatty acid composition and oxidative stability of chicken tissues was investigated. Chicks were fed on diets containing (g/kg): fresh sunflower oil (FSO) 40, heated sunflower oil (HSO) 40 or heated sunflower oil (40) supplemented with alpha-tocopheryl acetate (HSE) to a similar alpha-tocopherol concentration as the FSO diet. Concentrations of alpha-tocopherol in tissues of chicks fed on HSO and HSE were significantly lower than those of chicks fed on FSO. Significant correlations were observed between plasma alpha-tocopherol concentration and the alpha-tocopherol concentrations of other tissues (r > or = 0.67, P < 0.005) and between log plasma alpha-tocopherol and plasma thiobarbituric acid-reacting substances (TBARS) concentrations (r -0.851, P < 0.001). The concentrations of TBARS in tissues of chicks fed on the various diets were generally very similar before stimulation of peroxidation with Fe-ascorbate. Susceptibility of tissues to Fe-ascorbate-induced lipid peroxidation was increased by feeding HSO. Supplementation with alpha-tocopheryl acetate reduced susceptibility to lipid oxidation to varying degrees, depending on the tissue. The results suggest that chronic ingestion of oxidized lipids may compromise free-radical-scavenging activity in vivo by depleting alpha-tocopherol in the gastrointestinal tract, or possibly in plasma and other tissues.

Influence of heated vegetable oils and alpha-tocopheryl acetate supplementation on alpha-tocopherol, fatty acids and lipid peroxidation in chicken muscle.

1. Chicks were fed on diets containing fresh, heated or alpha-tocopheryl acetate-supplemented heated vegetable oils. The effects on alpha-tocopherol status, and on the fatty acid composition and oxidative stability of thigh and breast muscle were determined. 2. Plasma alpha-tocopherol was significantly correlated with alpha-tocopherol concentrations in thigh and breast muscle. 3. The fatty acid profiles of muscle lipids reflected dietary fatty acid composition. 4. The consumption of heated sunflower and linseed oils reduced alpha-tocopherol status, altered fatty acid composition of muscle lipids and increased susceptibility of muscle to lipid oxidation. 5. Supplementation of diets containing heated oils with alpha-tocopheryl acetate resulted in some alleviation of these effects. 6. The results indicate that caution should be exercised in the use of thermally oxidised oils in poultry diets if undesirable changes in composition and oxidative stability of carcase lipids are to be avoided.

Influence of dietary alpha-tocopherol on tocopherol concentrations in chick tissues.

1. The effect of feeding alpha-tocopherol (5 to 180 micrograms/g diet) for 24 days on the concentrations of alpha-tocopherol in various chicken tissues was investigated. 2. Tissue alpha-tocopherol concentrations responded to dietary intake in the order: heart congruent to lung greater than liver greater than thigh muscle greater than brain, and in all cases the relationship between the concentrations of dietary and tissue alpha-tocopherol was highly significant (0.997 less than or equal to r less than or equal to 1). 3. Plasma alpha-tocopherol concentration appears to be a good index (r greater than or equal to 0.910, P less than 0.001) of alpha-tocopherol status of lung, liver and heart.