Concerted Phenotypic Flexibility of Avian Erythrocyte Size and Number in Response to Dietary Anthocyanin Supplementation

Background: Endurance flight impose substantial oxidative costs on the avian oxygen delivery system. In particular, the accumulation of irreversible damage in red blood cells can reduce the capacity of blood to transport oxygen and limit aerobic performance. Many songbirds consume large amounts of anthocyanin-rich fruit, which is hypothesized to reduce oxidative costs, enhance post-flight regeneration, and enable greater aerobic capacity. While their antioxidant benefits appear most straightforward, the effects of anthocyanins on blood composition remain so far unknown. We fed thirty hand-raised European starlings (Sturnus vulgaris) two semisynthetic diets (with or without anthocyanin supplement) and manipulated the extent of flight activity in a wind tunnel (daily flying or non-flying for over two weeks) to test for their interactive effects on functionally important haematological variables. Results: Supplemented birds had on average 15% more and 4% smaller red blood cells compared to non-supplemented individuals and these diet effects were independent of flight manipulation. Haemoglobin content was 7% higher in non-supplemented flying birds compared to non-flying birds, while similar haemoglobin content was observed among supplemented birds that were flown or not. Neither diet nor flight activity influenced haematocrit. Conclusion: The concerted adjustments suggest that supplementation generally improved antioxidant protection in blood, which could prevent the excess removal of cells from the bloodstream and may have several implications on the oxygen delivery system, including improved gas exchange and blood flow. The flexible haematological response to dietary anthocyanins may also suggest that free-ranging species preferentially consume anthocyanin-rich fruits for their natural blood doping, oxygen delivery-enhancement effects

The Energy Savings-Oxidative Cost Trade- Off for Migratory Birds During Endurance Flight

Elite human and animal athletes must acquire the fuels necessary for extreme feats, but also contend with the oxidative damage associated with peak metabolic performance. Here, we show that a migratory bird with fuel stores composed of more omega-6 polyunsaturated fats (PUFA) expended 11% less energy during long-duration (6 hr) flights with no change in oxidative costs; however, this short-term energy savings came at the long-term cost of higher oxidative damage in the omega-6 PUFA-fed birds. Given that fatty acids are primary fuels, key signaling molecules, the building blocks of cell membranes, and that oxidative damage has long-term consequences for health and ageing, the energy savings-oxidative cost trade-off demonstrated here may be fundamentally important for a wide diversity of organisms on earth

Mitochondrial metabolism in blood more reliably predicts whole-animal energy needs compared to other tissues

Summary: Understanding energy metabolism in free-ranging animals is crucial for ecological studies. In birds, red blood cells (RBCs) offer a minimally invasive method to estimate metabolic rate (MR). In this study with European starlings Sturnus vulgaris, we examined how RBC oxygen consumption relates to oxygen use in key tissues (brain, liver, heart, and pectoral muscle) and versus the whole organism measured at basal levels. The pectoral muscle accounted for 34%–42% of organismal MR, while the heart and liver, despite their high mass-specific metabolic rate, each contributed 2.5%–3.0% to organismal MR. Despite its low contribution to organismal MR (0.03%–0.04%), RBC MR best predicted organismal MR (r = 0.70). Oxygen consumption of the brain and pectoralis was also associated with whole-organism MR, unlike that of heart and liver. Overall, our findings demonstrate that the metabolism of a systemic tissue like blood is a superior proxy for organismal energy metabolism than that of other tissues

Dietary Antioxidants Attenuate the Endocrine Stress Response during Long-duration Flight of a Migratory Bird

Glucocorticoids (GCs) are metabolic hormones that promote catabolic processes, which release stored energy and support high metabolic demands such as during prolonged flights of migrating birds. Dietary antioxidants (e.g. anthocyanins) support metabolism by quenching excess reactive oxygen species produced during aerobic metabolism and also by activating specific metabolic pathways. For example, similar to GCs’ function, anthocyanins promote the release of stored energy, although the extent of complementarity between GCs and dietary antioxidants is not well known. If anthocyanins complement GCs functions, birds consuming anthocyanin-rich food can be expected to limit the secretion of GCs when coping with a metabolically challenging activity, avoiding the exposure to potential hormonal detrimental effects. We tested this hypothesis in European starlings (Sturnus vulgaris) flying in a wind tunnel. We compared levels of corticosterone, the main avian GC, immediately after a sustained flight and at rest for birds that were fed diets with or without an anthocyanin supplement. As predicted, we found (i) higher corticosterone after flight than at rest in both diet groups and (ii) anthocyanin-supplemented birds had less elevated corticosterone after flight than unsupplemented control birds. This provides novel evidence that dietary antioxidants attenuate the activation of the HPA axis (i.e. increased secretion of corticosterone) during long-duration flight

Environmental Cues and Dietary Antioxidants Affect Breeding Behavior and Testosterone of Male European Starlings (Sturnus vulgaris).

Environmental cues, such as photoperiod, regulate the timing of major life-history events like breeding through direct neuroendocrine control. Less known is how supplementary environmental cues (e.g., nest sites, food availability) interact to influence key hormones and behaviors involved in reproduction, specifically in migratory species with gonadal recrudescence largely occurring at breeding sites. We investigated the behavioral and physiological responses of male European starlings to the sequential addition of nest boxes and nesting material, green herbs, and female conspecifics and how these responses depend on the availability of certain antioxidants (anthocyanins) in the diet. As expected, cloacal protuberance volume and plasma testosterone of males generally increased with photoperiod. More notably, testosterone levels peaked in males fed the high antioxidant diet when both nest box and herbal cues were present, while males fed the low antioxidant diet showed no or only a muted testosterone response to the sequential addition of these environmental cues; thus our results are in agreement with the oxidation handicap hypothesis. Males fed the high antioxidant diet maintained a constant frequency of breeding behaviors over time, whereas those fed the low antioxidant diet decreased breeding behaviors as environmental cues were sequentially added. Overall, sequential addition of the environmental cues modulated physiological and behavioral measures of reproductive condition, and dietary antioxidants were shown to be a key factor in affecting the degree of response to each of these cues. Our results highlight the importance of supplementary environmental cues and key resources such as dietary antioxidants in enhancing breeding condition of males, which conceivably aid in attraction of high quality females and reproductive success

Season, Anthocyanin Supplementation, and Flight Training Have Mixed Effects on the Antioxidant System of Migratory European Starlings

Migratory birds engage in 2 periods of endurance flight annually as they travel between summer breeding and overwintering grounds, and such endurance flights likely incur oxidative costs. These costs may differ between fall and spring migration, especially for females who must prepare for breeding and egg laying in spring. The objective of this study of a migratory bird was to test proposed hypotheses about how key components of the female’s antioxidant system differ in response to flight training in the fall and spring and to dietary antioxidant supplementation. We hand raised female European Starlings (Sturnus vulgaris) and fed them either a diet supplemented with dietary anthocyanins or a diet without added anthocyanins. We flew females in a wind tunnel for 15 days during fall and spring migration seasons and measured over time oxidative lipid damage (d-ROMs) and 3 components of the antioxidant system: nonenzymatic antioxidant capacity (OXY), uric acid, and glutathione peroxidase (GPx) activity. Prior to flight training, OXY and oxidative damage were lower in females during spring compared with fall, and females fed a low-antioxidant diet had consistently higher circulating uric acid. GPx activity decreased more in spring immediately after a long-duration flight. Females fed a high-antioxidant diet had a greater decrease in OXY after the 15-day flight training. Flight-trained females had higher circulating uric acid than untrained females immediately after the longest-duration flight and decreased GPx activity after the 15-day flight training. In sum, females upregulated enzymatic and nonenzymatic endogenous antioxidants in spring, and females fed a diet with less antioxidants appear to compensate by increasing circulating uric acid. Our findings emphasize the important role of dietary antioxidants for birds during migration, and similar flights in fall and spring likely represent distinct oxidative challenges in the life history of female birds

Flight Training And Dietary Antioxidants have Mixed Effects On The Oxidative Status of Multiple Tissues In A Female Migratory Songbird

Birds, like other vertebrates, rely on a robust antioxidant system to protect themselves against oxidative imbalance caused by energy-intensive activities such as flying. Such oxidative challenges may be especially acute for females during spring migration, since they must pay the oxidative costs of flight while preparing for reproduction; however, little previous work has examined how the antioxidant system of female spring migrants responds to dietary antioxidants and the oxidative challenges of regular flying. We fed two diets to female European starlings, one supplemented with a dietary antioxidant and one without, and then flew them daily in a windtunnel for two weeks during the fall and spring migration periods. We measured the activity of enzymatic antioxidants (GPx, SOD, CAT), non-enzymatic antioxidant capacity (ORAC), and markers of oxidative damage (protein carbonyls and lipid hydroperoxides) in four tissues: pectoralis, leg, liver, and heart. Dietary antioxidants affected enzymatic antioxidant activity and lipid damage in the heart, non-enzymatic antioxidant capacity in the pectoralis, and protein damage in leg muscle. In general, birds fed less antioxidants appear to incur increased oxidative damage while upregulating non-enzymatic and enzymatic antioxidant activity, though these effects were strongly tissue-specific. We also found trends for diet x training interactions for enzymatic antioxidant activity in the heart and leg. Flight-training may condition the antioxidant system of females to dynamically respond to oxidative challenges, and females during spring migration may shift antioxidant allocation to reduce oxidative damage