Stopovers Serve Physiological Recovery in Migratory Songbirds.

AbstractMigrating birds perform extreme endurance exercise when flying. This shifts the balance between the production of reactive oxygen species and the antioxidant defense system toward the former, potentially generating oxidative damages. In between migratory flights, birds make stopovers, where besides accumulating fuel (mainly fats), they are assumed to rest and recover from the strenuous flight. We performed a series of studies on both temporarily caged (northern wheatears) and free-flying (northern wheatears and European robins) migrants to investigate whether migrants recover during stopover by decreasing the amount of oxidative lipid damage (malondialdehyde [MDA]) and/or increasing the total nonenzymatic antioxidant capacity (AOX). In caged wheatears, MDA decreased within a single day. These birds were able to simultaneously accumulate considerable amounts of fuel. Also, in the free-flying wheatears, there was a decrease in MDA during stopover; however, this process seemed incompatible with refueling. The reason for this difference could relate to constraints in the wild that are absent in caged birds, such as food limitation/composition and locomotor activity. In the robins, there was a near significant decrease in MDA concentration in relation to how long the birds were already at stopover, suggesting that this species also physiologically recovers during stopover. AOX did not change during stopover in either of the wheatear studies. For the robins, however, uric acid-corrected AOX declined during stopover. Our results show that during stopover, migrating birds rapidly reduce oxidative lipid damage, thereby likely recovering their physiological state. In addition to the commonly accepted function of refueling, stopovers thus probably serve physiological recovery.

Recovery of constitutive immune function after migratory endurance flight in free-living birds.

Strenuous physical activity can negatively affect constitutive innate immune function (CIF), the always present first line of defence against pathogens. CIF is non-specific, and thus vital when encountering novel pathogens. A lowered CIF likely increases the risk of infection and disease. Migratory birds engage in truly extreme physical activity during their endurance flights, however, little is known about how they deal with the negative impact this has on their immune function. By collecting both between- and within-individual data we show, for the first time, that free-flying migratory birds can recover several parameters of CIF during stopovers, which are stationary periods in between migratory flights. With this, we provide an important piece of the puzzle on how migrating birds cope with the physiological challenges they face on their biannual journeys. Furthermore, our study stresses the importance of migratory stopovers beyond fuel accumulation.

Migrating birds rapidly increase constitutive immune function during stopover.

Migratory flight is physiologically highly demanding and has been shown to negatively affect multiple parameters of constitutive immune function (CIF), an animal's first line of physiological defence against infections. In between migratory flights, most birds make stopovers, periods during which they accumulate fuel for the next flight(s). Stopovers are also commonly thought of as periods of rest and recovery, but what this encompasses is largely undefined. Here, we show that during stopover, northern wheatears Oenanthe oenanthe, a long-distance migratory bird, can rapidly increase constitutive innate immune function. We caught and temporarily caged birds under ad libitum food conditions at a stopover site in autumn. Within 2 days, most birds significantly increased complement activity and their ability to kill microbes. Changes in immune function were not related to the birds' food intake or extent of fuel accumulation. Our study suggests that stopovers may not only be important to refuel but also to restore immune function. Additionally, the increase in CIF could help migrating birds to deal with novel pathogens they may encounter at stopover sites.

Diel variation in corticosterone and departure decision making in migrating birds.

Animals usually show distinct periods of diel activity and non-activity. Circulating baseline levels of glucocorticoid hormones (corticosterone and cortisol) often peak just before or at the transition from the non-active to the active period of the day. This upregulation of glucocorticoids may function to mobilize stored energy and prepare an animal for increased activity. Usually, the alternation of active and non-active periods is highly predictable; however, there is one group of animals for which this is not always the case. Many otherwise diurnal birds show nocturnal activity during the migration seasons. Nocturnal migratory flights are alternated with stopover periods during which the birds refuel and rest. Stopovers vary in length, meaning that nocturnal migrants are inactive in some nights (when they continue their stopover) but extremely active in other nights (when they depart and fly throughout the night). This provides an ideal natural situation for testing whether glucocorticoids are upregulated in preparation for an increase in activity, which we used in this study. We found that in northern wheatears (Oenanthe oenanthe), corticosterone levels peaked in the few hours before sunset in birds departing from stopover that night, and, importantly, that this peak was absent in birds continuing stopover. This indicates that corticosterone is upregulated in the face of an increase in energy demands, underlining corticosterone's preparative metabolic function (energy mobilization). The timing of upregulation of corticosterone also gives a first insight in when during the day nocturnally migrating birds decide whether or not to resume migration.

An exception to the rule: Captivity does not stress wild migrating northern wheatears.

Wild animals typically suffer from stress when brought into captivity. This stress is characterized by elevated circulating glucocorticoid levels and weight loss. We here describe for the first time a case where a wild animal, the long-distance migrating northern wheatear, does not show signs of stress when caged. We captured these birds on a stopover site during their spring migration and caged them individually with ad libitum access to food and water. The birds were divided into four groups and were blood-sampled immediately in the field, a few hours after caging, one day after caging, or three days after caging, respectively. From these blood-samples we determined circulating corticosterone level. Food intake and body mass were also monitored. We found that, with very few exceptions, corticosterone levels were low and did not differ among the groups. Accordingly, almost all birds consumed huge quantities of food and substantially increased their body mass. Together these results clearly show that caging does not result in indications of stress in wild migrating northern wheatears. Confinement-specific conditions such as restricted movement normally stress animals. We suggest migratory birds may not perceive such conditions as stressors due to their hyperphagic state, a notion that requires further testing.

Ghrelin, corticosterone and the resumption of migration from stopover, an automated telemetry study.

The spectacular natural phenomenon of avian migration is evidently shaped by physical factors, but we know little about the underlying physiological regulation. This contrast is especially apparent for the process of departure on a migratory flight. The decision to resume migration is shaped by a suite of departure cues from innate rhythms, and intrinsic and extrinsic factors. It currently appears that these departure cues are translated into actual departure by the hormone corticosterone, but other hormones may play a role too and probably interact with corticosterone. We captured this concept here by investigating the role of the hormone ghrelin and its interaction with corticosterone for the departure decision in a migratory songbird. Ghrelin functions as an appetite-regulating hormone. It has also been suggested to be involved in the regulation of departure by upregulating corticosterone in migrants ready to depart, and by facilitating the breakdown of lipids to fuel migratory flight. We measured plasmatic ghrelin and corticosterone levels in migrating common blackbirds (Turdus merula) at an autumnal stopover site, and determined their departure timing with the use of a fully-automated radio-telemetry system. Against our expectations, ghrelin level was not correlated with the birds' lipid stores or with corticosterone level. Furthermore, departure likelihood and nocturnal departure time were not associated with ghrelin levels. Our study thus does not support the idea that ghrelin is involved in the regulation of departure from stopover, at least not in common blackbirds. We discuss possible reasons for the lack of confirmation of our expectations.

Cues, corticosterone and departure decisions in a partial migrant.

Most migrating birds make multiple stopovers to fuel and/or rest. The decision to resume migration from stopover is based on various cues, such as time within the season and wind conditions. There are hints that the strength of these departure cues shapes corticosterone level, which in its turn appears to regulate the timing of departure. We here provide results that very strongly indicate that indeed departure cues jointly shape corticosterone level of migrants at stopover. We compared corticosterone level between migrating and sedentary common blackbirds (Turdus merula) sampled simultaneously at the same location during autumn migration. As expected, in migrating individuals corticosterone level was positively associated with time within the season and with current wind conditions. The latter was only apparent in adult birds and not in 1st year migrants, thus matching the observation that 1st year autumnal migrants are less wind selective than adults. In contrast to the migrants, in sedentary blackbirds these "cues" did not explain variation in corticosterone level. Furthermore, stopover departure seemed more likely and to occur earlier in the night in migrants with high corticosterone level. Our unique comparative study thus supports the newly developed concept that corticosterone mediates between departure cues and stopover departure timing in avian migrants.

Corticosterone and timing of migratory departure in a songbird.

Bird migration entails replenishing fuel stores at stopover sites. There, individuals make daily decisions whether to resume migration, and must also decide their time of departure. Variation in departure timing affects the total time required to complete a migratory journey, which in turn affects fitness through arrival time at the breeding and wintering grounds. It is well established that stopover departure decisions are based on cues from innate rhythms, intrinsic factors and extrinsic factors. Yet, virtually nothing is known about the physiological mechanism(s) linking these cues to departure decisions. Here, we show for a nocturnal migratory songbird, the northern wheatear (Oenanthe oenanthe), that baseline corticosterone levels of birds at stopover increased both over the migratory season and with wind assistance towards the migratory destination. Corticosterone in turn predicted departure probability; individuals with high baseline corticosterone levels were more likely to resume migration on a given night. Corticosterone further predicted the departure time within the night, with high baseline levels being associated with early departures. These novel findings indicate that corticosterone may be mediating between departure cues and the timing of departure from a stopover site, which is a major step towards understanding the hormonal control of animal migration.