Polyphenol compounds in the chicken/animal diet: from the past to the future.

Animal feed provides a range of antioxidants that help the body building an integrated antioxidant system responsible for a prevention of damaging effects of free radicals and products of their metabolism. Vitamin E is considered to be the main chain-breaking antioxidant located in the membranes and effectively protecting them against lipid peroxidation. Recently, various polyphenol compounds, especially flavonoids, have received substantial attention because of their antioxidant activities in various in vitro systems. However, it was shown that flavonoid compounds are poorly absorbed in the gut and their concentrations in target tissues are too low to perform an effective antioxidant defences. The aim of the present paper is to review existing evidence about possible roles of various plant extracts provided with the diet in animal/poultry nutrition with a specific emphasis to their antioxidant activities.

Selenoproteins and maternal nutrition.

Selenium (Se) is an essential trace element of fundamental importance to health due to its antioxidant, anti-inflammatory and chemopreventive properties attributed to its presence within at least 25 selenoproteins (Sel). Sel include but not limited to glutathione peroxidases (GPx1-GPx6), thioredoxin reductases (TrxR1-TrxR3), iodothyronine deiodinases (ID1-ID3), selenophosphate synthetase 2 (SPS2), 15-kDa Sel (Sel15), SelH, SelI, SelK, SelM, SelN, SelO, SelP, SelR, SelS, SelT, SelV, SelW, as well as the 15-kDa Sel (Fep15), SelJ and SelU found in fish. In this review, we describe some of the recent progress in our understanding of the mechanisms of Sel synthesis. The impact of maternal Se intake on offspring is also discussed. The key regulatory point of Sel synthesis is Se itself, which acts predominantly at post-transcriptional levels, although recent findings indicate transcriptional and redox regulation. Maternal nutrition affects the performance and health of the progeny. Both maternal and offspring Se supplementations are essential for the antioxidant protection of the offspring. Prenatal Se supplementation provides an effective antioxidant system that is already in place at the time of birth while, postnatal Se supplementation becomes the main determinant of progeny Se status after the first few days of progeny life.

[Effect of selenium-containing supplements on the indices of specific immunity and nonspecific resistance in chicken].

The deficit of selenium is related to immunity worsening. Selenium improves an immune answer and raises disease resistance. The aim of our work was to study the influence of organic selenium in comparison with inorganic selenium on the factors of unspecific resistance and state of humoral immunity of chicken-broilers.

An analysis of pre- and post-hatching maternal effects mediated by carotenoids in the blue tit.

Maternal effects increase phenotypic plasticity in offspring traits and may therefore facilitate adaptation to environmental variability. Carotenoids have been hypothesized to mediate costs of reproduction in females as well as maternal effects. However, assessing potential transgenerational and population consequences of environmental availability of carotenoids requires a better understanding of mechanisms of maternal effects mediated by these antioxidant pigments. Manipulating dietary availability of carotenoids to egg-laying female blue tits and subsequently cross-fostering nestlings between female treatments allowed us to specifically investigate the relative importance of maternal effects through egg carotenoids and through post-hatching care mediated by antioxidants in females. Nestling body size and mass and plasma antioxidants were not significantly affected by pre- or post-hatching maternal effects mediated by antioxidants, although both types of maternal effects in interaction explained the variation in growth, as measured by wing length. Development of the ability to mount a cell-mediated immune response as well as its temporal dynamics was influenced by both pre- and post-hatching maternal effects, with an advantage to nestlings originating from, or reared by, carotenoid-supplemented females. In addition, nestlings reared by carotenoid-fed females had a lower blood sedimentation rate, indicating that they may have been less infected than nestlings from controls. Finally, prehatching maternal effects in interaction with nestling plasma carotenoid levels affected the development of carotenoid-based plumage. Maternal effects mediated by carotenoids may thus act as a proximate factor in development and phenotypic plasticity in traits associated with nestling fitness, such as immune response and ability to metabolize and use antioxidants, and ultimately participate in the evolution of phenotypic traits.

Maternal effects and the evolution of brain size in birds: overlooked developmental constraints.

A central dogma for the evolution of brain size posits that the maintenance of large brains incurs developmental costs, because they need prolonged periods to grow during the early ontogeny. Such constraints are supported by the interspecific relationship between ontological differences and relative brain size in birds and mammals. Given that mothers can strongly influence the development of the offspring via maternal effects that potentially involve substances essential for growing brains, we argue that such effects may represent an important but overlooked component of developmental constraints on brain size. To demonstrate the importance of maternal effect on the evolution of brains, we investigated the interspecific relationship between relative brain size and maternal effects, as reflected by yolk testosterone, carotenoids, and vitamins A and E in a phylogenetic study of birds. Females of species with relatively large brains invested more in eggs in terms of testosterone and vitamin E than females of species with small brains. The effects of carotenoid and vitamin A levels on the evolution of relative brain size were weaker and non-significant. The association between relative brain size and yolk testosterone was curvilinear, suggesting that very high testosterone levels can be suppressive. However, at least in moderate physiological ranges, the positive relationship between components of maternal effects and relative brain size may imply one aspect of developmental costs of large brains. The relationship between vitamin E and relative brain size was weakened when we controlled for developmental mode, and thus the effect of this antioxidant may be indirect. Testosterone-enhanced neurogenesis and vitamin E-mediated defence against oxidative stress may have key functions when the brain of the embryo develops, with evolutionary consequences for relative brain size.

Comparative analysis of apoptosis induction by selenium compounds in human lymphoblastic leukemia MT-4 cells.

AIM: The precise mechanisms of apoptosis induced by various selenium compounds are not well understood. Therefore, the apoptogenic activity of two inorganic selenium compounds, sodium selenite and sodium selenate, in human T-cell acute lymphoblastic leukemia MT-4 cells was compared focusing on their effects on cell cycle progression and activation of proapoptotic Bax protein. METHODS: Apoptosis and cell cycle distribution of MT-4 cells exposed to inorganic selenium compounds was assessed by flow cytometry upon propidium iodide staining. Bax expression was analyzed by flow cytometry of cells labeled with anti-Bax monoclonals. Extent of DNA damage was assessed by Comet analysis. RESULTS: Sodium selenite induced apoptosis in dose-dependent mode starting from concentrations of 10 microM, while sodium selenate was much less toxic inducing apoptotic cell death only at 300 microM. Sodium selenite but not sodium selenate caused a slight arrest in G2/M phase of cell cycle. The cytotoxicity of sodium selenite was accompanied by DNA damaging effects visualized in DNA comet assay. Nevertheless, the drastic increase in Bax flow cytometry intensity was evident only in selenite-induced apoptosis. CONCLUSION: Sodium selenite induced apoptosis is accompanied by increased Bax expression.

Effect of organic selenium on turkey semen quality during liquid storage.

The aim of the present study was to evaluate the effect of dietary organic selenium on the turkey semen during storage. Twenty males (BUT, Big 6, 40 weeks of age) were divided into control (n=10) and experimental group (n=10). The turkeys in the both groups were fed with a commercial diet containing 0.1 ppm Se in the form of sodium selenite. The experimental birds were additionally supplied with 0.3 ppm organic Se in the form Sel-Plex (Alltech, Inc.). After 30 days of feeding, the semen samples were collected twice a week for the 3 weeks of the study and diluted 1+1(v/v) with TUR-2 diluent, and stored in a water bath (+10 to 15 degrees C) for 6 h. The percentage of motile spermatozoa, the sperm viability (live/dead spermatozoa), total lipids, phospholipids and total cholesterol were assessed in fresh and stored semen. The fertilizing ability of semen was assessed by artificial insemination of 30 hens per group with dose containing 200x10(6) spermatozoa weekly. After 6 h of semen storage, the motility of spermatozoa decreased significantly in the control group (by 8.7 relative percent, P<0.05) and only by four relative percent (P>0.05) in experimental group reflecting a protective effect of dietary Se supplementation. The proportion of live spermatozoa was higher in fresh semen and significantly lower in stored semen. The positive effect of Se supplementation was observed on the lipid composition of stored semen: the concentration of the total lipids and phospholipids in the seminal plasma from control group significantly increased, while in the experimental group remained constant. Better semen integrity in the experimental group was associated with an improved fertilizing ability of spermatozoa: the fertility rate of stored spermatozoa in the control group was 88%, while in the experimental group was 90.5%.

Effects of carotenoids from lucerne, marigold and tomato on egg yolk pigmentation and carotenoid composition.

1. The effects of various sources of natural carotenoids (Px alfalfa concentrate, tomato powder and marigold extract) as feed additives in quail diets on egg yolk pigmentation and carotenoid composition were investigated. 2. Adult Japanese quail were fed one of 5 different diets for 23 d: three diets each contained Px alfalfa concentrate (PX) or tomato powder (TP) or marigold extract (MG), one diet contained marigold extract and tomato powder (MG + TP) and a control diet (wheat/barley based) was low in carotenoid. All products were added at a rate of 2%, apart from marigold extract which was added at a rate of 0.2%. 3. Visual assessment of yolk colour (Roche colour fan) showed a stabilised yolk colour of 1.6, 7.7, 8.5, 8.8 and 10.6 for the control, PX, TP, MG and MG + TP treatments, respectively. 4. The total carotenoid concentration of the egg yolks were 2.2, 22.4, 4.1, 39.0 and 37.7 microg/g for the experimental groups fed the following diets: control, PX, TP, MG and MG + TP, respectively. Deposition of retinol, alpha-tocopherol and gamma-tocopherol in the egg yolk was unaffected by treatments. 5. Lutein was shown to be the major carotenoid in the egg yolk, comprising 1.65, 17.97, 2.03, 31.14 and 28.57 microg/g in control, PX, TP, MG and MG + TP, respectively. Inclusion of TP in the quail diet resulted in lycopene transfer to the egg. 6. It was concluded that, in comparison with the control group, there was an increase in the yolk concentrations of lutein, zeaxanthin, lycopene and beta-carotene in eggs produced by female quail fed diets supplemented with natural carotenoids.

Effects of supplementing broiler breeder diets with organoselenium compounds and polyunsaturated fatty acids on hatchability.

The effects of supplementing broiler breeder diets with polyunsaturated fatty acids (PUFA) and organoselenium compounds on fertility, hatchability, and the weight of 1-d-old chicks was assessed. Prepeak (23 wk) and peak (27 wk) production breeders were fed 1 of 4 diets: a wheat-based commercial breeder diet with 55 g/kg of either soybean oil (SO) or fish oil (FO), but no added Se (only that originating from feed ingredients), and each diet with added Se as Sel-Plex (SO + Se, FO + Se). The diets were designed to contain <0.1 mg/kg of Se and about 0.5 mg/kg of Se for the nonsupplemented (no added Se) and the supplemented diets, respectively. The Se concentration of the eggshell of the hatching egg was measured. The concentration of Se, PUFA, and total lipid content of the brain and liver of the 1-d-old chick was determined. The number of fertile eggs increased, embryonic mortality decreased, and hatchability increased as hen age increased from 23 to 27 wk. The Se concentration in the eggshell and the brain and liver of 1-d-old chicks was higher in the high-Se treatments com pared with the concentration in the low-Se treatments. Fish oil inclusion in the breeder diet increased embryonic mortality in wk 3 of incubation and reduced both hatchability and 1-d-old chick weight in hens of both ages. The addition of Se to the FO diets ameliorated some of these adverse effects, because chicks hatched from eggs laid by 23-wk-old breeders of the FO + Se treatment were heavier than those receiving the FO treatment. The Se concentration in the brain and liver of chicks from the FO hens was higher than that in chicks from the SO hens. The concentration of docosahexaenoic fatty acid was higher in the liver of chicks from the SO + Se treatment compared with that of chicks from the SO treatment, indicating possible protective effects of Se. Hatchability was decreased by increased PUFA and was higher in 27-wk-old compared with 23-wk-old breeders.

Maternal organo-selenium compounds and polyunsaturated fatty acids affect progeny performance and levels of selenium and docosahexaenoic acid in the chick tissues.

The effects of supplementing broiler breeder diets with polyunsaturated fatty acids (PUFA) and organo-Se compounds on the levels of Se and PUFA in chick tissues and on chick performance were assessed. Prepeak (23 wk) and peak (27 wk) production broiler breeders were fed 1 of 4 diets: a wheat-based commercial diet with soybean oil or fish oil but no added Se, and each diet with added Se as Sel-Plex (soybean oil + Se, fish oil + Se; Alltech Inc, Nicholasville, KY). The diets were designed to contain less than 0.1 mg of Se/kg and about 0.5 mg/kg for the nonsupplemented and the supplemented diets, respectively. As-hatched chicks from the 4 parental treatments were fed a nutritionally high quality diet (ME = 12.57 MJ/kg; CP = 228.7 g/kg) or a low quality diet (ME = 10.28 MJ/kg; CP = 182.8 g/kg), resulting in 8 dietary treatments. Performance was better and mortality lower in chicks from 27-wk-old breeders compared with those from 23-wk-old breeders. Fish oil in the maternal diet increased progeny mortality and reduced chick body mass at hatch. Body mass at 7 and 14 d posthatch was lower in chicks fed the low quality diet compared with chicks fed the high quality diet. At hatch, and for up to 14 d posthatch, chicks from hens fed diets high in PUFA had higher concentrations of docosahexaenoic acid (DHA) in the brain and liver compared with chicks hatched from hens fed diets low in PUFA. The DHA content of the tissues of chicks from breeders fed diets supplemented with Se was higher than that in chicks from breeders fed unsupplemented diets. Even after 14 d of being fed a diet with identical levels of Se, chicks hatched from parents fed diets high in Se had higher tissue Se concentrations than those hatched from parents fed diets low in Se. Supplementation of the maternal diet of chicks with organo-Se appears to enhance the DHA concentration of the chick brain, which may improve brain function.

Multiple pathways of maternal effects in black-headed gull eggs: constraint and adaptive compensatory adjustment.

We investigated in the black-headed gull whether female deposition of antioxidants and immunoglobulins (enhancing early immune function), and testosterone (suppressing immune function and increasing early competitive skills) correlate suggesting that evolution has favoured the mutual adjustment of different pathways for maternal effects. We also took egg mass, the position of the egg in the laying sequence and offspring sex into account, as these affect offspring survival. Yolk antioxidant and immunoglobulin concentrations decreased across the laying order, while yolk testosterone concentrations increased. This may substantially handicap the immune defence of last-hatched chicks. The decrease in antioxidant levels was greater when mothers had a low body mass and when the increase in testosterone concentrations was relatively large. This suggests that female black-headed gulls are constrained in the deposition of antioxidants in last-laid eggs and compensate for this by enhanced testosterone deposition. The latter may be adaptive since it re-allocates the chick's investment from costly immune function to growth and competitive skills, necessary to overcome the consequences of hatching late from an egg of reduced quality.

Supplementation of the maternal diet with tomato powder and marigold extract: effects on the antioxidant system of the developing quail.

The effects of natural dietary carotenoid supplementation of the maternal diet (tomato powder and marigold extract) on transfer to the egg yolk and on the development of the antioxidant system of the young quail liver in early postnatal life were investigated. Sixty Japanese quail (Coturnix coturnix japonica) were allocated to four treatment groups, each with three replicates consisting of four females and one male each. The quail were fed on one of four different diets for 23 d, each of them based on a low carotenoid, wheat/barley-based control diet. Tomato powder and marigold extract were added at rates of 20 and 2 g/kg to treatments 2 and 3, respectively. Marigold extract and tomato powder were also used in combination in treatment 4 at 2 g marigold + 20 g tomato powder/kg of diet. At 20 weeks of age, 60 eggs from each treatment were collected and placed in an incubator. After hatching, d-old quail from each group were reared (under standard commercial conditions) up to 14 d of age. They were fed on a low-carotenoid commercial diet. After hatch, at 1, 7 and 14 d, the livers of five young quail from each treatment were assessed for total carotenoid concentration and carotenoid profile. Results indicated that lycopene is transferred from the feed to the egg yolk and further to the liver of the developing embryo. Elevated carotenoid concentration in the egg yolk and correspondingly in the liver of newly hatched quail remains significant during first week posthatch. Lutein and lycopene did not affect vitamin E concentration in the egg yolk or liver of the newly hatched quail. A combination of increased concentrations of lycopene and lutein in the egg yolk results in elevated concentrations of coenzyme Q in the liver of the newly hatched quail.

Effect of organic selenium in quail diet on its accumulation in tissues and transfer to the progeny.

1. The aim of the present study was to investigate the effects on the eggs and hatchlings (up to 2 weeks post-hatch) of feeding a relatively large amount of so-called organic selenium to breeder quail. 2. Two groups of quail (3 families in each group consisting of 4 females and 1 male) were formed at the beginning of their reproductive period. The quail were fed on a commercial maize-based diet containing 0.096 mg/kg feed-derived selenium (Se), supplemented with 0.2 mg/kg selenite (control group) or 0.5 mg/kg organic selenium in the form of Sel-Plex (Alltech Ltd, USA) for 6 months. Eggs were collected at 6 months of age and Se in the egg yolk, egg white and shell was analysed. Five quail at 1, 7 and 14 d post-hatch were killed to provide samples of liver, brain, breast and leg muscles for Se analysis. After egg collection for analysis and incubation, adult quail were killed and liver, kidney, lung, brain, breast and leg muscles were collected for Se analyses. 3. Inclusion of high doses (0.5 mg/kg) of organic Se in the quail diet was associated with a significant increase in Se concentration in all tissues studied of adult quail as well as in egg yolk, egg albumin and eggshell. 4. Increased Se concentration in the quail egg was associated with increased Se concentration in the liver, breast and leg muscles and brain of newly hatched quail. This difference was shown to be significant for 2 weeks post-hatch. Therefore, it has been suggested that the maternal effect of dietary selenium can be seen beyond the hatching time and more emphasis should be given to this effect in future. 5. It was shown that it is possible to produce Se-enriched quail meat and eggs by adding organic selenium to the diet.

Effects of supplementing broiler breeder diets with organic selenium and polyunsaturated fatty acids on egg quality during storage.

We assessed the effects of supplementation of broiler breeder diets with polyunsaturated fatty acids (PUFA) and organic Se on hatching egg quality during storage. Broiler breeders (n = 352) were fed 1 of 4 diets: SO (soybean oil with no added Se), SO with Se (SO+Se), fish oil (FO), or FO and Se (FO+Se). Eggs from prepeak (23 wk) and peak production (27 wk) breeders were stored for 14 d under typical (15 degrees C, 78% RH) conditions. Eggs were analyzed for albumen Haugh units (HU) and pH, shell characteristics, egg component weight, Se content, and fatty acid (FA) profile. The efficiency of transfer of Se into the egg was greater in low Se treatments compared with in high Se treatments. The majority of Se from the low Se treatments was deposited in the yolk, whereas in high Se treatments, the Se was deposited evenly in the yolk and albumen. In the low Se treatments, the Se content of yolk and albumen was lower than in the high Se treatments and decreased as age increased. Albumen HU decreased with increased storage, although high Se treatments had greater HU compared with low Se treatments. Eggs from the FO treatment were smaller with thinner eggshells compared with the SO treatment, but addition of Se to the FO diets ameliorated some of these effects. The egg yolk FA profile from the SO+Se treatment was altered compared with that from the SO treatment, perhaps due to interactions between dietary PUFA and Se-dependent enzyme systems. Hatching egg quality during storage was affected by dietary PUFA and Se content, as well as broiler breeder age.

Dietary fish and evening primrose oil with vitamin E effects on semen variables in cockerels.

Our aim was to determine the effect of n-3 (2%, wt/wt, fish oil rich diet) and n-6 (2%, wt/wt, evening primrose oil rich diet) fatty acid dietary supplementation and their combination with two concentrations of vitamin E (40 vs 200 mg/kg) on semen variables and on fatty acid and vitamin E profiles of spermatozoa in broiler breeders at 32, 42 and 52 weeks of age. The inclusion of fish oil in the cockerel diets increased the docosahexaenoic acid proportion in the sperm phospholipid fraction, which was almost threefold higher compared to the other two groups irrespective of vitamin E supplementation. In contrast, an increase in the proportion of total n-6 polyunsaturates, mainly 22:4n-6, was observed in the evening primrose oil group compared to the control only when the dietary content of vitamin E was increased to 200 mg/kg. Sperm concentration was decreased in the fish and evening primrose oil groups if vitamin E was 40 mg/kg, but such an effect was prevented in the fish, not the evening primrose oil group, by increasing the vitamin E to 200 mg. The proportion of motile spermatozoa was improved by the increased supplementation of vitamin E in all oil treatments.

Antioxidants, radiation and mutation as revealed by sperm abnormality in barn swallows from Chernobyl.

Reduced levels of antioxidants such as carotenoids and vitamins A and E can increase DNA damage caused by free radicals. Exposure to radiation has been proposed to reduce levels of antioxidants that are used for DNA repair and this reduction may be responsible for increased levels of mutation in radioactively contaminated areas. We test this hypothesis using field measures of antioxidants in blood, liver and eggs of the barn swallow Hirundo rustica while relating these to levels of mutation as reflected by the frequency of abnormal sperm. Antioxidant levels in blood, liver and eggs were reduced in Chernobyl, Ukraine, compared with an uncontaminated control area, and levels of antioxidants correlated negatively with levels of background radiation. The frequency of abnormal sperm was almost an order of magnitude higher in Chernobyl than in the control area and was negatively related to antioxidant levels in blood and liver. This is consistent with the hypothesis of a direct link between radiation and individual levels of antioxidants, suggesting that levels of mutation differ among individuals owing to individual differences in the abundance of antioxidants.

Effect of canthaxanthin content of the maternal diet on the antioxidant system of the developing chick.

1. Effects of canthaxanthin supplementation of the maternal diet on the antioxidant system of the developing chick were investigated. 2. Three hundred and twenty female broiler breeder birds were housed in one of 4 controlled environment rooms with 3 replicates for all treatments, with the exception of the control treatment of which there were 4 replicates. All birds received one of 5 diets: control low xanthophyll diet, or the same diet supplemented with 3, 6, 12 or 24 mg/kg canthaxanthin in the form of Carophyll Red. At 30 weeks of age 60 eggs from each of the 5 groups were incubated. At d 16 of the embryo development, at d 1 and d 7 posthatch tissue samples were collected and analysed by HPLC-based methods. 3. Canthaxanthin accumulation in the egg yolk was proportional to dietary content. Furthermore, at 12 to 24 mg/kg canthaxanthin was associated with an increase in gamma-tocopherol concentration in the egg yolk. Canthaxanthin was transferred from the egg yolk to the developing embryo and, as a result, its concentration in the liver of the embryo at 16 and in 1-d-old chicks was increased. Even at d 7 posthatch canthaxanthin concentration in the chicken liver was elevated. 4. Canthaxanthin supplementation of the maternal diet at 12 mg/kg was associated with an increased alpha-tocopherol concentration in the liver of 1-d-old chicks and resulted in decreased tissue susceptibility to lipid peroxidation. 5. Canthaxanthin supplementation at 6 to 24 mg/kg was also associated with a delay in alpha-tocopherol depletion from the liver for 7-d posthatch. As a result of the increased canthaxanthin and vitamin E concentrations in the liver of 7-d-old chicks, tissue susceptibility to lipid peroxidation decreased. 6. The results support an idea that dietary carotenoids can modulate antioxidant systems of the developing chicken.

Transfer of Vitamins E and A from yolk to embryo during development of the king penguin (Aptenodytes patagonicus).

Since the yolk lipids of the king penguin (Aptenodytes patagonicus) are rich in n-3 fatty acids, which are potentially susceptible to peroxidative damage, the yolk contents and yolk-to-embryo transfer of antioxidants and lipid-soluble vitamins were investigated under conditions of natural incubation in the wild. The concentration of vitamin E in the unincubated egg was 155 microg/g wet yolk, of which 88% was alpha-tocopherol and the rest was gamma-tocopherol. Vitamin A (2.9 microg/g) was present in the yolk entirely as retinol; no retinyl esters were detected. Throughout the latter half of the incubation period, vitamins E and A were taken up from the yolk into the yolk sac membrane (YSM) and later accumulated in the liver, with vitamin A being transferred in advance of vitamin E. In the YSM, vitamin A was present almost entirely as retinyl ester, indicating that the free retinol of the yolk is rapidly esterified following uptake. Retinyl esters were also the predominant form in the liver. The retinyl esters of the liver and YSM displayed different fatty acid profiles. At hatching, the brain contained relatively little vitamin E (4.7 microg/g) compared to the much higher concentration in the liver (482.9 microg/g) at this stage. Ascorbic acid was not detected in the yolk but was present at a high concentration in the brain at day 27 (404.6 microg/g), decreasing to less than half this value by the time of hatching. This report is the first to delineate the yolk-to-embryo transfer of lipid-soluble vitamins for a free-living avian species. The yolk fatty acids of the king penguin provide an extreme example of potential oxidative susceptibility, forming a basis for comparative studies on embryonic antioxidant requirements among species of birds whose yolk lipids differ in their degree of unsaturation.