The influences of Tylosine and licorice dietary supplementation in terms of the productive performance, serum parameters, egg yolk lipid profile, antioxidant and immunity status of laying Japanese quail under heat stress condition.

By reducing feed consumption, animals suffering from heat stress prefer to reduce their heat output. Heat exposure has also contributed to major adverse effects on the productive and reproductive performance of quails. Therefore, this research was intended to estimate the preventive function of licorice as a safe feed additive against the negative effects caused by heat stress conditions on laying quail productivity. A total number of 180 Japanese quail birds (120 females and 60 males), nine-weeks old were divided into five groups. Each group contained 36 birds in four replicates (nine birds) with completely randomized design. The dietary treatments were a basal diet without supplementation as control (T1), basal diet + 100 mg Tylosine kg-1 diet (T2), (T3), (T4) and (T5) fed basal diet + 250, 500 and 1000 mg licorice kg-1 diet, in respect. The results indicated that dietary supplementation with different feed additives had no significant effects on egg-laying rate, egg number, average egg weight, egg mass and feed conversion ratio compared with control. Also, different treatments showed no significant variations on serum IgG, total protein, globulin, albumin, creatinine, ALT and TAC and caused significant (P ≤ 0.05) improvement in IgM, AST, ALP, uric acid and MDA concentrations as compared to the control. Results indicated that total serum lipids, triglycerides, total cholesterol and LDL concentrations were significantly decreased due to different feed additives. However, HDL concentrations and HDL/LDL ratios were markedly increased by the other treatments than the control group. In addition, yolk total lipids were significantly (P ≤ 0.001) decreased with increasing licorice root powder's dietary levels compared with the control group. Also, a significant (P ≤ 0.01) reduction in egg yolk cholesterol level was observed in the group fed with 500 mg licorice compared to other treatments. In conclusion, fortified laying quail diets with licorice powder could be a useful strategy to alleviate adverse effects induced by heat stress as alternative to antibiotics on laying Japanese quail.

Effects of supplementation of dietary humate, probiotic, and their combination on performance, egg quality, and yolk fatty acid composition of laying hens.

This study was carried out to determine the effects of addition of humate, probiotic, and their combination into diets on performance, egg quality, and yolk fatty acid composition of hens during the second laying period. Lohmann LSL white layers (n = 192), 46 weeks of age, were randomly divided into 4 groups and fed with basal diet (control, C), 0.3% humate (H), 0.3% probiotic (P), 0.15% humate + 0.15% probiotic (HP) for 18 weeks. Feed consumption and egg production were determined daily, egg weight was measured biweekly, and body weights were recorded at the beginning and the end of the experiment. Also, 12 egg samples from each group were randomly collected to determine the egg quality every 30 days. Laying performance, yolk color, and fatty acid composition were significantly (P < 0.05 and P < 0.01) affected by addition of humate, probiotic, and their combination into diets of layers. The HP group had higher cracked egg yield and feed conversion ratio values than control and H and P groups. Except for egg yolk color, the other egg quality parameters such as shape index, shell strength, shell thickness, albumen index, yolk index, and Haught unit were not affected by treatment (P < 0.05 and P < 0.01). The egg yolks of treatment groups had less stearic acid than those of control group. In conclusion, supplementation of humate and probiotic into the diets of laying hens increased monounsaturated fatty acids in yolk and improved feed conversion ratio and egg yolk color.

Comparison of Antioxidants: The Limited Correlation between Various Assays of Antioxidant Activity.

The inhibitory effects a range of synthetic and natural antioxidants on lipid peroxidation of egg yolk and erythrocyte membranes induced by a free radical generator 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) was compared, with significant differences being found between both systems. When the protection by selected antioxidants against the effects of AAPH on erythrocytes (hemolysis, oxidation of hemoglobin and glutathione (GSH) and generation of reactive oxygen species (ROS)) was studied, most antioxidants were protective, but in some tests (oxidation of hemoglobin and GSH) some acted as prooxidants, inducing oxidation in the absence of AAPH and enhancing the AAPH-induced oxidation. These results demonstrate a diversified action of antioxidants in different systems and point to a need for careful extrapolation of any conclusions drawn from one parameter or experimental system to another.

Prenatal independent and combined effects of yolk vitamin E and corticosterone on embryo growth and oxidative status in the yellow-legged gull.

Variation in the concentration of antioxidants and hormones of maternal origin in the eggs of birds can have a profound influence on offspring phenotype both prenatally and postnatally. Egg maternal substances can have interacting effects, but experimental studies of the consequences of the combined variation in the egg concentration of such molecules are extremely rare, particularly as far as prenatal stages are concerned. We manipulated the yolk concentration of vitamin E and corticosterone, which are, respectively, the main antioxidant and the main glucocorticoid hormone in bird eggs, both independently and simultaneously, and we tested their separate and combined effects on growth and oxidative status in the liver and in the brain of yellow-legged gull (Larus michahellis) embryos. Egg supplementation of relatively large physiological doses of corticosterone depressed embryo growth (total body mass, tarsus length and liver mass), whereas administration of vitamin E in association with corticosterone restored normal growth. Vitamin E did not affect embryo growth when administered alone. We further analysed the independent and combined effects of vitamin E and corticosterone on liver and brain total antioxidant capacity, the concentration of reactive oxygen molecules and lipid peroxidation. Vitamin E significantly reduced liver total antioxidant capacity, while corticosterone depressed brain lipid peroxidation. Prenatal exposure to vitamin E and corticosterone appears to have antagonistic effects on body growth, although vitamin E is not limiting in yellow-legged gull eggs. In combination with the results of previous experiments on the same species applying smaller experimental doses or focusing on the postnatal rather than prenatal life stages, our findings indicate that the effects of a physiological increase in the egg concentration of these substances can be life stage and dose specific, implying that generalizing prenatal effects of egg compounds may not be feasible.

Oregano essential oil (Origanum vulgare) to feed laying hens and its effects on animal health.

This study evaluated the effect of oregano essential oil added to the feed of commercial laying hens. This research was focused on the analysis of biochemical changes linked to hepatic function, and protein and lipid metabolism. It was used 240 laying hens (59 weeks-old) distributed in a completely randomized design of six treatments (five repetitions with eight birds each). The experiments were constituted by a control treatment (CT) with the inclusion of zinc bacitracin and five treatments of oregano essential oil (OEO: 0, 50, 100, 150, and 200 mg.kg-1), respectively. After 28 days of feeding, an increase on serum levels of total proteins and globulins was observed on groups T150 and T200, as well as an increase on albumin levels on group CT. After 84 days of feeding, a significant reduction on total proteins and albumin was observed on group T200, as well as an increase in serum triglycerides. OEO at 200 mg.kg-1 increased globulin levels on day 28, which may be considered an effect in the inflammatory response, which increases serum immunoglobulins and proteins.

Performance and egg quality of laying hens fed with mineral sources and rosemary oil.

In order to evaluate the effect of rosemary oil and micro mineral sources on the performance and egg quality of laying hens, 288 hens were used and distributed in a completely randomized design using a 2x3 factorial (mineral sources x rosemary oil) with six different diets and six replications. Diets were formulated containing inorganic or organic minerals, with or without added rosemary oil (100 and 200 mg kg-1). The treatments showed interaction for average egg weight, yolk color, albumen and yolk percentage, with significant results from the use of organic minerals compared to inorganic minerals. The addition of 200 mg kg-1 rosemary oil in diets improved laying rate, egg mass, specific gravity, number of pores on the shell when compared to other treatments. In conclusion, the use of 200 mg kg-1 of rosemary oil improves the performance of red laying hen eggs. The association between organic minerals and 100 mg kg-1 rosemary oil in laying hen diets increases yolk color and percentage of albumen. The use of organic minerals is superior to inorganic minerals as to improve the quality of eggs, increasing average egg weight, yolk color and percentage of albumen.

Effects of dietary fat source on lutein, zeaxanthin and total carotenoids content of the egg yolk in laying hens during the early laying period.

1. The study was conducted to investigate the effects of different dietary levels of an unsaturated or saturated fat source and the effects of different dietary energy levels in laying hen diets on the carotenoid content of the egg yolk. 2. Seventy-two ISA Brown laying hens aged 22 weeks old were allocated, for a 10 week period, to three treatments: control diet (HE group) with a 3.4% supplemented fat containing 2.1% soybean and 1.3% palm oil as fat sources and a ratio of unsaturated to saturated fatty acid (u/s) of 3.33; a lowered energy diet with 2.4% supplemented fat containing 1.4% soybean and 1.0% palm oil and u/s of 3.41 (LE+high u/s); a lowered energy diet with 2.4% supplemented fat containing 0.5% soybean and 1.9% palm oil and a u/s of 2.39 (LE+low u/s). A marigold plant extract supplement was used as a source of lutein. 3. Performance parameters (feed consumption, feed conversion, body weight), egg production rate and egg quality parameters were similar between treatments. Dietary fat digestibility at the middle of the study period was not different. Egg yolk total fat content was similar in all treatments at the middle and at the end of the study period. 4. Lutein, zeaxanthin, cis-lutein and total carotenoids content were significantly lower in eggs produced from the LE+low u/s group compared to those from the HE and the LE+high u/s groups (P < 0.01 for all parameters). 5. It was concluded that feeding laying hens with a diet containing 1% less supplemented fat and a lower u/s ratio compared to a control diet and to a diet with 1% less supplemented fat with a higher u/s ratio resulted in a significant reduction of carotenoid expression in the egg yolk at the end of experimental period. Performance and egg quality parameters were not affected by treatments.

Influence of citrus flavonoids on laying hen performance, inflammatory immune response, egg quality and yolk oxidative stability.

1. The objective of this study was to investigate the effects of dietary supplementation with natural flavonoids (naringin and hesperidin) on laying hens' performance, cellular immunity and egg quality parameters. 2. A total of 72 individually caged laying hens were allocated into 1 of 6 treatment groups: a control (C) group that was fed with a basal diet and groups that were offered the same diet further supplemented with hesperidin at 0.75 g/kg (E1), or 1.5 g/kg (E2), or naringin at 0.75 g/kg (N1), or 1.5 g/kg (N2) or α-tocopheryl acetate at 0.2 mg/kg (VE) for 67 d. 3. Supplementation with naringin or hesperidin did not affect the performance and egg quality (P > 0.05) apart from an improvement in the yolk colour that was more orange in naringin and hesperidin groups in comparison to the controls (P-linear < 0.05). Egg yolk and plasma cholesterol levels were not affected by citrus flavonoids (P-linear > 0.05). 4. Inflammatory immune response, measured by phytohaemagglutinin skin test (PHA), was suppressed in laying hens that were fed with either naringin (P-linear < 0.05) or hesperidin (P < 0.05). Egg yolk oxidative stability was improved from the 4th d after naringin or hesperidin supplementation. This beneficial effect was comparable to that of α-tocopheryl acetate and was observed in eggs that were stored for up to 120 d. 5. In conclusion, naringin and hesperidin may favourably prolong the shelf life of eggs, appear to possess anti-inflammatory properties and could improve the yolk colour without any side effects on the performance or egg quality traits.

Flaxseed and dried tomato waste used together in laying hens diet.

The aim of this study was to investigate the effects of simultaneous supplementation of laying hens with dietary sources of n-3 polyunsaturated fatty acids (PUFA) and carotenoids on egg quality, fatty acids and carotenoid profile of the egg yolk and on feed and yolk lipid peroxidation. A 6-week experiment was carried out with 53-week old laying hens (96 Tetra SL) assigned to a control and three treatment groups supplemented with 5% flaxseeds and different levels of dried tomato waste (DTW, 2.5%, 5.0% and 10.0%). Hens from the groups supplemented with 5% and 7.5% DTW had a significantly lower average daily feed intake and laying percentage as compared to the control. Increased doses of dietary DTW enhanced yolk Roche colour score in direct correlation with the enrichment of egg yolk in carotenoids but decreased their transfer efficiency from feed to egg. After 4 weeks, egg yolk from hens fed with 5% flaxseeds and 7.5% DTW had increased lutein and zeaxanthin levels (by 29% and 24%, respectively) and the colour score was 3.5 fold higher compared to the control group. As a result of the dietary supplementation with flaxseed, the n-3 fatty acid content was 3.1-3.7-fold higher in egg yolk compared with the control and the n-6/n-3 ratio decreased from 18.3 (control) to 4.1-5.4 in supplemented diets. Dietary supplementation with 5% DTW effectively prevented lipid oxidation of eggs enriched with n-3 PUFA, but the increase in DTW content depressed the absorption and deposition of n-3 PUFA in egg yolk.

Effect of yolk corticosterone on begging in the yellow-legged gull.

Behavioral lateralization is widespread across vertebrates. The development of lateralization is affected by both genetic and environmental factors. In birds, maternal substances in the egg can affect offspring lateralization via activational and/or organizational effects. Corticosterone affects the development of brain asymmetry, suggesting that variation in yolk corticosterone concentration may also influence post-natal behavioral lateralization, a hypothesis that has never been tested so far. In the yellow-legged gull (Larus michahellis), we increased yolk corticosterone concentration within physiological limits and analyzed the direction of lateralization of hatchlings in reverting from supine to prone position ('RTP' response) and in pecking at dummy parental bills to solicit food provisioning ('begging' response). We found that corticosterone treatment negatively affected the frequency of begging and it may cause a slight leftward lateralization. However, the direction of lateralization of the RTP response was not affected by corticosterone administration. Thus, our study shows a maternal effect mediated by corticosterone on a behavioral trait involved in parent-offspring communication during food provisioning events. The findings on lateralization are not conclusive due to the weak effect size but provide information for further ecological and evolutionary studies, investigating mechanisms underlying the development of lateralization.

Independent and combined effects of egg pro- and anti-oxidants on gull chick phenotype.

Oviparous mothers transfer to their eggs components that have both independent and combined effects on offspring phenotype. The functional interaction between egg components, such as antioxidants and hormones, suggests that a change in the concentration of one component will have effects on offspring traits that depend on the concentration of other interacting components. However, the combined effects of variation in different egg components are virtually unknown. Bird eggs contain vitamin E, a major antioxidant, and also maternal corticosterone. The independent consequences of variation in the egg concentrations of these compounds for offspring phenotype are largely unknown and no study has investigated their combined effects. We manipulated the concentration of vitamin E and corticosterone in the eggs of the yellow-legged gull (Larus michahellis) by administering a physiological (2 s.d.) dose both independently and in combination. We tested for an effect on chick post-natal growth, plasma antioxidant capacity (TAC) and oxidative compounds (TOS). Separate administration of vitamin E or corticosterone caused a reduction in body mass relative to controls, whereas the combined administration of the two compounds reversed their negative effects. These results suggest that maternal egg components, such as antioxidants and steroid hormones, interact and mothers must balance their concentrations in order to achieve optimal offspring phenotype. The functional relationship between vitamin E and corticosterone is corroborated by the observation of positive covariation between these compounds.

High post-thaw survival of ram sperm after partial freeze-drying.

BACKGROUND: Recrystallization damages occur when a frozen sample is held at high subzero temperatures and when the warming process is too slow. METHODS: In this work, ram semen diluted in two different concentrations of sugar solutions (Lyo A consisted of 0.4 M sorbitol and 0.25 M trehalose, and the second, Lyo B composed of 0.26 M sorbitol and 0.165 M trehalose) in egg yolk and Tris medium were compared after freezing 10 µL samples to: (1) - 10, - 25, and - 35 °C and thawing. (2) Freezing to - 10 and - 25 °C, holding for 1 h and then thawing, and (3) freezing to - 10 and - 25 °C and drying for 1 h at these temperatures at a vacuum of 80 mTorr, prior thawing. For drying, we used a new freeze-drying apparatus (Darya, FertileSafe, Israel) having a condensation temperature below - 110 °C and a vacuum pressure of 10-100 mTorr that is reached in less than 10s. RESULTS: Results showed that samples in Lyo B solution frozen at - 25 °C had significantly higher sperm motility in partially freeze-dried samples than frozen samples (46.6 ± 2.8% vs 1.2 ± 2.5%, P < 0.001). Moreover, partially dried samples in Lyo B showed higher motility than Lyo A at - 25 °C (46.6 ± 2.8% vs 35 ± 4%). Cryomicroscopy and low-temperature/low-pressure environmental scanning electronic microscope demonstrated that the amount of the ice crystals present in partially dried samples was lower than in the frozen samples. CONCLUSION: Holding the sperm at high subzero temperatures is necessary for the primary drying of cells during the freeze-drying process. Rapid freeze-drying can be achieved using this new device, which enables to reduce recrystallization damages.

Pharmacokinetics of tapentadol in laying hens and its residues in eggs after multiple oral dose administration.

1. The aim of the study was to evaluate the pharmacokinetics (PKs) of tapentadol (TAP), a novel opioid analgesic, in laying hens after intravenous (IV) and oral (PO) administration and to quantify the concentrations of TAP residues in eggs. 2. Twenty healthy laying hens were divided into three groups: A (n = 6), B (n = 6) and C (n = 8). The study was conducted in two phases. Groups A and B received TAP by IV and PO routes at the dose of 1 and 5 mg/kg, respectively. 3. No visible adverse effects were observed after administration of the drug. TAP plasma concentrations were detectable up to 4 h following administration. Following IV administration, TAP plasma concentrations were only higher than the minimal effective concentration (148 ng/ml) reported for humans for 1 h. After single PO administration, plasma concentrations of TAP would not conform to software algorithms and the PK parameters were not calculated. TAP concentration following multiple PO doses at 5 mg/kg for 5 d was found to be higher and more persistent (12 h vs. 7 h) in yolk compared with albumen. 4. This is the first PK study on the novel atypical opioid TAP in laying hens. Further studies are required to investigate the analgesic efficacy and actual effective plasma concentration of TAP in this species.

Effects of extruded flaxseed on layer performance, nutrient retention and yolk fatty acid composition.

1. This study was conducted to determine the effects of feeding an extruded flaxseed (EF) on layer performance, apparent total tract nutrient retention (ATTR) and egg yolk fatty acid concentrations. 2. Seventy-two White Leghorn laying hens (58-week-old; three per cage) were randomly allotted to one of four dietary treatments: 0%, 7.5%, 15.0% and 22.5% of EF-supplemented diets for 8 weeks. 3. Supplementation with EF had no effect on feed intake, egg production, feed conversion ratio and egg weight. Egg components (yolk, albumen and shell percentages) were similar among treatments, except that shell percentage was greater for layers fed 22.5% EF than those fed 7.5% and 15% EF. The ATTR of dry matter and organic matter were highest for 0% and 7.5% EF, intermediate for 15% EF and lowest for 22.5% EF. Similar reductions on ATTR of crude protein and nitrogen-corrected apparent metabolisable energy were observed for layers fed 22.5% EF relative to those fed 0% or 7.5% EF. 4. Feeding EF at 7.5%, 15.0% and 22.5% of the diet markedly increased (by 92%, 198% and 271%, respectively) egg yolk concentrations of n-3 polyunsaturated fatty acid (PUFA) and reduced saturated fatty acid and n-6 PUFA concentrations. 5. It was concluded that omega-3 labelled eggs (300 mg/60 g of egg) may be produced with low (7.5% of diet) levels of dietary EF without compromising egg production parameters. However, feeding moderate to high levels of EF (i.e. 15% and 22.5% EF) may reduce total tract nutrient and energy utilisation.

Coadaptation of offspring begging and parental provisioning: A role for prenatal maternal effects?

Recent studies on birds have shown that offspring begging and parental provisioning covary at the phenotypic level, which is thought to reflect genetic correlations. However, prenatal maternal factors, like yolk testosterone, may also facilitate parent-offspring coadaptation via their effects on offspring begging and development. In fact, maternal effects are thought to adjust offspring phenotype to the environmental conditions they will experience after birth, which are in turn strongly dependent on the levels of parental provisioning. Using cross-fostering experiments in canaries, we tested the role of maternal effects on parent-offspring coadaptation from two different approaches. First, we analyzed whether females deposit yolk testosterone in relation to their own or their partner's prospective parental provisioning, measured as the rate of parental feeding to foster nestlings. Second, we investigated whether females deposit yolk testosterone in relation to costs they incurred when raising a previous brood, as this likely impinges on their capacity to provide parental care in the near future. However, from the results of both experiments we have no evidence that canary females deposit yolk testosterone in order to match offspring begging to the levels of care they and/or their partners provide. We therefore found no evidence that yolk testosterone facilitates parent-offspring coadaptation. In addition, our results suggest that the functional consequences of yolk testosterone deposition may relate to hatching asynchrony since it primarily varied with egg laying order.

Higher growth rate and gene expression in male zebra finch embryos are independent of manipulation of maternal steroids in the eggs.

Sexual dimorphism in prenatal development is widespread among vertebrates, including birds. Its mechanism remains unclear, although it has been attributed to the effect of maternal steroid hormones. The aim of this study was to investigate how increased levels of steroid hormones in the eggs influence early embryonic development of male and female offspring. We also asked whether maternal hormones take part in the control of sex-specific expression of the genes involved in prenatal development. We experimentally manipulated hormones' concentrations in the egg yolk by injecting zebra finch females prior to ovulation with testosterone or corticosterone. We assessed growth rate and expression levels of CDK7, FBP1 and GHR genes in 37h-old embryos. We found faster growth and higher expression of two studied genes in male compared to female embryos. Hormonal treatment, despite clearly differentiating egg steroid levels, had no effect on the sex-specific pattern of the embryonic gene expression, even though we confirmed expression of receptors of androgens and glucocorticoids at such an early stage of development. Thus, our study shows high stability of the early sex differences in the embryonic development before the onset of sexual differentiation and indicates their independence of maternal hormones in the egg.

Effects of Curcuma longa rhizome powder on egg quality, performance and some physiological indices of laying hens fed different levels of metabolizable energy.

BACKGROUND: High-energy diets of laying hens may improve roductive performance, although some negative effects may also appear with respect to egg quality and physiological parameters. Curcuma longa rhizome powder (CRP) has beneficial effects on health indices of the birds through antioxidant and antimicrobial properties, especially when the birds experience nutritional stress. RESULTS: Increasing dietary CRP enhanced egg quality by improving eggshell thickness and hardness but decreasing yolk cholesterol content (P < 0.05). The best feed conversion ratio was obtained in birds fed high-apparent metabolizable energy (AME) diets supplemented with 2.0 g kg-1 CRP (P < 0.05). Although increasing dietary AME elevated the serum concentration of triglycerides (P < 0.05) and enzymatic activities of alanine aminotransferase and aspartate aminotransferase (P < 0.05), dietary inclusion of CRP alleviates the blood levels of these enzymes (P < 0.01). Low level of dietary CRP boosted the immune responses to Newcastle virus (P < 0.01) and sheep red blood cells (P < 0.05) antigens but decreased the heterophil to lymphocyte ratio (P < 0.05). Inclusion of at least 2.0 g kg-1 CRP in the diet of laying decreased Escherichia coli enumerations in the ileal content (P < 0.01) and improved villus height, crypt depth and goblet cell numbers (P < 0.05). CONCLUSION: An improvement in the productive performance of laying hens fed high-energy diets might be associated with decreasing health indices and product quality, which could potentially be amended by nutritional modifications such as incorporating medicinal herbs in the feed. © 2016 Society of Chemical Industry.

Effects of experimentally manipulated yolk thyroid hormone levels on offspring development in a wild bird species.

Maternal effects are a crucial mechanism in a wide array of taxa to generate phenotypic variation, thereby affecting offspring development and fitness. Maternally derived thyroid hormones (THs) are known to be essential for offspring development in mammalian and fish models, but have been largely neglected in avian studies, especially in respect to natural variation and an ecological context. We studied, for the first time in a wild species and population, the effects of maternally derived THs on offspring development, behavior, physiology and fitness-related traits by experimental elevation of thyroxine and triiodothyronine in ovo within the physiological range in great tits (Parus major). We found that elevated yolk TH levels had a sex-specific effect on growth, increasing male and decreasing female growth, relative to controls, and this effect was similar throughout the nestling period. Hatching or fledging success, motor coordination behavior, stress reactivity and resting metabolic rate were not affected by the TH treatment. We conclude that natural variation in maternally derived THs may affect some offspring traits in a wild species. As this is the first study on yolk thyroid hormones in a wild species and population, more such studies are needed to investigate its effects on pre-hatching development, and juvenile and adult fitness before generalizations on the importance of maternally derived yolk thyroid hormones can be made. However, this opens a new, interesting avenue for further research in the field of hormone mediated maternal effects.

Dietary antioxidants and flight exercise in female birds affect allocation of nutrients to eggs: how carry-over effects work.

Physiological challenges during one part of the annual cycle can carry over and affect performance at a subsequent phase, and antioxidants could be one mediator of trade-offs between phases. We performed a controlled experiment with zebra finches to examine how songbirds use nutrition to manage trade-offs in antioxidant allocation between endurance flight and subsequent reproduction. Our treatment groups included (1) a non-supplemented, non-exercised group (control group) fed a standard diet with no exercise beyond that experienced during normal activity in an aviary; (2) a supplemented non-exercised group fed a water- and lipid-soluble antioxidant-supplemented diet with no exercise; (3) a non-supplemented exercised group fed a standard diet and trained to perform daily endurance flight for 6 weeks; and (4) a supplemented exercised group fed an antioxidant-supplemented diet and trained to perform daily flight for 6 weeks. After flight training, birds were paired within treatment groups for breeding. We analyzed eggs for lutein and vitamin E concentrations and the plasma of parents throughout the experiment for non-enzymatic antioxidant capacity and oxidative damage. Exercised birds had higher oxidative damage levels than non-exercised birds after flight training, despite supplementation with dietary antioxidants. Supplementation with water-soluble antioxidants decreased the deposition of lipid-soluble antioxidants into eggs and decreased yolk size. Flight exercise also lowered deposition of lutein, but not vitamin E, to eggs. These findings have important implications for future studies of wild birds during migration and other oxidative challenges.

Glucose metabolism and gene expression in juvenile zebrafish (Danio rerio) challenged with a high carbohydrate diet: effects of an acute glucose stimulus during late embryonic life.

Knowledge on the role of early nutritional stimuli as triggers of metabolic pathways in fish is extremely scarce. The objective of the present study was to assess the long-term effects of glucose injection in the yolk (early stimulus) on carbohydrate metabolism and gene regulation in zebrafish juveniles challenged with a high-carbohydrate low-protein (HC) diet. Eggs were microinjected at 1 d post-fertilisation (dpf) with either glucose (2 M) or saline solutions. Up to 25 dpf, fish were fed a low-carbohydrate high-protein (LC) control diet, which was followed by a challenge with the HC diet. Survival and growth of 35 dpf juveniles were not affected by injection or the HC diet. Glucose stimulus induced some long-term metabolic changes in the juveniles, as shown by the altered expression of genes involved in glucose metabolism. On glycolysis, the expression levels of hexokinase 1 (HK1) and phosphofructokinase-6 (6PFK) were up-regulated in the visceral and muscle tissues, respectively, of juveniles exposed to the glucose stimulus, indicating a possible improvement in glucose oxidation. On gluconeogenesis, the inhibition of the expression levels of PEPCK in fish injected with glucose suggested lower production of hepatic glucose. Unexpectedly, fructose-1,6-bisphosphatase (FBP) expression was induced and 6PFK expression reduced by glucose stimulus, leaving the possibility of a specific regulation of the FBP-6PFK metabolic cycle. Glucose metabolism in juveniles was estimated using a [¹4C]glucose tracer; fish previously exposed to the stimulus showed lower retention of [¹4C]glucose in visceral tissue (but not in muscle tissue) and, accordingly, higher glucose catabolism, in comparison with the saline group. Globally, our data suggest that glucose stimulus at embryo stage has the potential to alter particular steps of glucose metabolism in zebrafish juveniles.