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.

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.

Endocrine regulation of migratory departure from stopover: Evidence from a longitudinal migratory restlessness study on northern wheatears.

Most migrating birds make stopovers to replenish fuel stores. The decision to resume migration from stopover to a large extent shapes the temporal organization of migration. This decision is known to be shaped by a suite of intrinsic and extrinsic factors such as the bird's fuel stores and current weather conditions. However, how departures from stopover are physiologically regulated is largely unknown. We here present data that strongly indicate that corticosterone, a hormone with a stimulatory effect on locomotion, acts as a mediator between fuel stores and departure from stopover. In migrating northern wheatears (Oenanthe oenanthe) temporarily caged at stopover, we observed a positive relationship between the change in fuel stores and the concurrent change in glucocorticoid metabolite (GCM) levels measured in the birds' droppings. We also found a positive relationship between the change in GCM levels and the change in the intensity of nocturnal migratory restlessness. As in northern wheatears nocturnal migratory restlessness is an accurate proxy for stopover departure likelihood, our results indicate that corticosterone mediates between fuel stores and the decision to resume migration. Our unique longitudinal study represents a considerable advance in our understanding of the endocrine regulation of avian migration.

Nocturnal departure timing in songbirds facing distinct migratory challenges.

Most migratory songbirds travel between their breeding areas and wintering grounds through a series of nocturnal flights. The timing of their departures defines the potential flight duration and thus the distance covered during a migratory night. Yet, migratory songbirds show substantial variation in their nocturnal departure timing. With this study, we aim to assess whether the respective challenges of the migration route, namely its distance and nature, help to explain this variation. At a stopover site, we caught Northern Wheatears (Oenanthe oenanthe) of two subspecies that differ in distance and nature of their onward migration route in spring, but not in autumn. We determined the start of their nocturnal migratory restlessness during short-term captivity, and radiotracked their nocturnal departure timing after release in both migration seasons. Northern Wheatears started their nocturnal migratory restlessness earlier when facing a long remaining migration distance and an extended sea barrier in spring. Individual departure directions generally affected the nocturnal departure timing with early departures being directed towards the respective migratory destination. In spring, this pattern was predominantly found in birds carrying relatively large fuel stores, but was absent in lean birds. At the same time, birds facing a short remaining migration distance and no extended sea barrier strongly reacted to relatively large fuel stores by an early start of nocturnal migratory behaviour (migratory restlessness and departure timing), whereas this reaction was not found in birds facing a long remaining migration distance and sea barrier. These results suggest that the basic diel schedule of birds' migratory activity is adapted to the onward migration route. Further, they suggest that birds adjust their behavioural response, that is start of nocturnal migratory behaviour, to fuel stores in relation to their impending migratory challenges. This is a substantial step in understanding variation of nocturnal departure timing and its adjustments in migratory songbirds. Further, it emphasizes the importance of interpreting birds' nocturnal migratory behaviour in the respective ecological context.

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.

Endocrine regulation of fueling by hyperphagia in migratory birds.

To support migratory endurance flight, birds accumulate large amounts of fat by hyperphagia (fueling). Whereas the factors influencing migrants' motivation to fuel are well described, the physiological mechanism regulating fueling is largely unknown. Hormones are likely involved and arguably the best studied with respect to food intake and fueling is corticosterone. Corticosterone has a permissive effect, as blocking the hormone's actions prohibits efficient fueling. There are no indications, though that corticosterone stimulates fueling, and some studies even observed negative correlations between corticosterone level and food intake and speed of fueling. The latter is unexpected as slow fueling could reduce the overall speed of migration. To test the causality of these negative correlations, I non-invasively increased circulating corticosterone levels in captive migrants and determined its effect on food intake and fuel accumulation. Neither food intake nor fuel accumulation differed between corticosterone-treated and control-treated individuals. This indicates that corticosterone does not hamper food intake and fueling during stopovers, nor does it stimulate these processes. Promising alternative candidates for the regulation of migratory hyperphagia are 'appetite regulating' hormones secreted by the adipose tissue, gut, or gastro-intestinal tract. The advance of next-generation sequencing will facilitate a bottom-up approach when investigating these.

How do energy stores and changes in these affect departure decisions by migratory birds? A critical view on stopover ecology studies and some future perspectives.

In birds, accumulating energy is far slower than spending energy during flight. During migration, birds spend, therefore, most of the time at stopover refueling energy used during the previous flight. This elucidates why current energy stores and actual rate of accumulating energy are likely crucial factors influencing bird's decision when to resume migration in addition to other intrinsic (sex, age) and extrinsic (predation, weather) factors modulating the decision within the innate migration program. After first summarizing how energy stores and stopover durations are generally determined, we critically review that high-energy stores and low rates of accumulating energy were significantly related to high departure probabilities in several bird groups. There are, however, also many studies showing no effect at all. Recent radio-tracking studies highlighted that migrants leave a site either to resume migration or to search for a better stopover location, so-called "landscape movements". Erroneously treating such movements as departures increases the likelihood of type II errors which might mistakenly suggest no effect of either trait on departure. Furthermore, we propose that energy loss during the previous migratory flight in relation to bird's current energy stores and migration strategy significantly affects its urge to refuel and hence its departure decision.

Migratory common blackbirds have lower innate immune function during autumn migration than resident conspecifics.

Animals need a well-functioning immune system to protect themselves against pathogens. The immune system, however, is costly and resource trade-offs with other demands exist. For migratory animals several (not mutually exclusive) hypotheses exist. First, migrants reduce immune function to be able to allocate resources to migration. Second, migrants boost immune function to cope with more and/or novel pathogens encountered during migration. Third, migrants reallocate resources within the immune system. We tested these hypotheses by comparing baseline immune function in resident and migratory common blackbirds (Turdus merula), both caught during the autumn migration season on the island of Helgoland, Germany. Indices of baseline innate immune function (microbial killing capacity and haptoglobin-like activity) were lower in migrants than in residents. There was no difference between the groups in total immunoglobulins, a measure of baseline acquired immune function. Our study on a short-distance avian migrant supports the hypothesis that innate immune function is compromised during migration.