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Birds have been observed to have dietary preferences for unsaturated fatty acids during migration. Polyunsaturated fatty acids (PUFAs) may increase the exercise performance of migrant birds; however, PUFAs are also peroxidation prone and might therefore incur increased costs in terms of enhanced oxidative damage in migratory individuals. To shed light on this potential constraint, we analyzed plasma fatty acid (FA) composition and estimated the unsaturation index as a proxy for susceptibility to lipid peroxidation of migrants and residents of the partially migratory common blackbird (Turdus merula) at a stopover site during autumn migration. As predicted, migrant birds had higher relative and absolute levels of PUFAs compared to resident birds. This included the strictly dietary ω‐3 PUFA α‐linolenic acid, suggesting a dietary and/or storage preference for these FAs in migrants. Interestingly, the FA unsaturation index did not differ between migrants and residents. These findings suggest a mechanism where birds alter their levels of metabolic substrate without simultaneously increasing the susceptibility of the substrate to lipid peroxidation. In summary, our results are in line with the hypothesis that increased exercise performance during migration might be constrained by oxidative stress, which is manifested in changes in the composition of key FAs to retain the unsaturation index constant despite the increased levels of peroxidizable PUFAs., Migrating blackbirds, Turdus merula, have higher plasma levels of exercise‐enhancing polyunsaturated fatty acids compared to resident conspecifics at the same stopover site. High levels of such unsaturated fatty acids may incur costs in terms of increased lipid peroxidation, and hence oxidative stress. Yet, the fatty acid unsaturation index did not differ between migrants and residents, suggesting physiological mechanisms in migrants to mitigate this potential cost.

Most migrating birds need to stopover in between flights in order to refuel. Lately, additional purposes of stopover have been suggested, including physiological recovery from metabolically demanding migratory flight. One apparently unavoidable, but harmful physiological effect of migratory flight is increased oxidative damage to lipids and proteins. We here, for the first time, tested whether migrating birds are able to reduce their oxidative damage during stopover. To be able to collect longitudinal data on a large number of individual birds, we temporarily caged wild northern wheatears, a long-distance migrant which does not suffer stress when caged during migration. Around noon on the first and third day at stopover, the birds were blood-sampled to determine malondialdehyde (MDA) concentration, a commonly used marker of oxidative damage to lipids. We found that MDA concentrations significantly decreased during stopover, a result unchanged when correcting for the peroxidizability of the substrate. The extent of the decrease was unrelated to the amounts of food consumed or of fuel accumulated. Our findings support the hypothesis that stopovers serve reduction of oxidative damage, warranting re-thinking of how birds accomplish their migrations. They also highlight the need to include physiological recovery as a driver of the (temporal) organization of migration. A free Plain Language Summary can be found within the Supporting Information of this article. (Less)

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.

In many animals, catabolic and anabolic periods are temporally separated. Migratory birds alternate energy expenditure during flight with energy accumulation during stopover. The size of the energy stores at stopover affects the decision to resume migration and thus the temporal organization of migration. We now provide data suggesting that it is not only the size of the energy stores per se that may influence migration scheduling, but also the physiological consequences of flying. In two subspecies of the northern wheatear Oenanthe oenanthe, a long-distance migrant, estimated energy stores at a stopover during autumn migration were positively related with both constitutive innate and acquired immune function, and negatively related with oxidative damage to lipids. In other words, migrants’ physiological condition was associated with their energetic condition. Although time spent at stopover before sampling may have contributed to this relationship, our results suggest that migrants have to trade-off the depletion of energy stores during flight with incurring physiological costs. This will affect migrants’ decisions when to start and when to terminate a migratory flight. The physiological costs associated with the depletion of energy stores may also help explaining why migrants often arrive at and depart from stopover sites with larger energy stores than expected. We propose that studies on the role of energy stores as drivers of the temporal organization of (avian) migration need to consider physiological condition, such as immunological and oxidative states.

Stopover periods in between migratory flights determine the speed of migration and as such may affect birds’ fitness. The decision to leave a stopover site is affected by, for example, energy stores and weather conditions. We investigated whether immunological state influences autumn stopover departure decisions in Northern Wheatear (Oenanthe oenanthe), a nocturnal long-distance migrant. We measured three parameters of constitutive immune function: microbial killing capacity, haptoglobin, and total immunoglobulins. None of them explained variation in departure probability, minimum stopover duration, or departure time within the night. Thus, we did not find an effect of the birds’ immunological state on stopover departure decisions.

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.

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.

Migration is energetically demanding and physiologically challenging. Migrating birds, for example, need to boost their antioxidant defenses to defeat the pro‐oxidants produced during high energetic activity. The enhanced antioxidant defense possibly withdraws limited resources (e.g., energy or micronutrients) from other physiological functions, such as immune defense. Such a trade‐off might not occur outside the migration seasons or in resident individuals. Here, we investigate whether there is a negative relationship between innate immune function and antioxidant defense by sampling both migrating and resident blackbirds (Turdus merula) at the same location during the same period of the annual cycle. We show that in migrating blackbirds microbial killing capacity (BKA), an integrative measure of baseline innate immune function was negatively correlated with total nonenzymatic antioxidant capacity. In contrast, in resident conspecifics, sampled at the same time and location, these two physiological measures were not correlated. This suggests that migrating birds trade off innate immune function and antioxidant defense. Furthermore, and likely a consequence of this trade‐off, in migrant blackbirds BKA was positively correlated with oxidative damage to lipids. In resident blackbirds BKA and degree of lipid oxidation were uncorrelated. The mechanism and currencies of the supposed trade‐off are currently unknown, but energetic investments or micronutrients are likely candidates. Future experimental studies could provide more conclusive evidence for this trade‐off; yet, our results open up a new level of thinking about the physiological costs of migration.

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.

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.

BackgroundSongbirds following distinct migration strategies (e.g. long- vs. short- to medium-distance migrants) often differ in their speed of migration during autumn and, thus, are assumed to face different time constraints. During migration, most songbird species alternate migratory flights with stopover periods. Many of them restrict these migratory flights to the night, i.e., they are nocturnal migrants. At stopover, nocturnal migrants need to select a specific night (night-to-night decision) and time of night (within-night decision) to resume migration. These departure decisions, which largely determine the speed of migration, are jointly affected by a set of intrinsic and extrinsic factors, i.e., departure cues. Here we aim to assess whether the set of intrinsic and extrinsic factors and the magnitude of their respective effects on stopover departure decisions differs between nocturnally migrating songbird species, depending on their migration strategy and associated time constraints.MethodsWe radio-tracked migrating Northern Wheatears (Oenanthe oenanthe; long-distance migrant), European robins (Erithacus rubecula) and Common Blackbirds (Turdus merula; both medium-distance migrants) during autumn stopover and analysed their night-to-night and within-night departure timing in relation to intrinsic and extrinsic factors.ResultsSpecies generally differed in their departure timing on both temporal scales, with shortest stopovers and earliest nocturnal departures in the long-distance migrant. Some factors, such as day of year, fuel load, cloud cover and crosswind, had consistent effects on stopover departure decisions in all three species. However, species differed in the effects of tailwind assistance, change in atmospheric pressure and air temperature on their stopover departure decisions. Whereas night-to-night decisions were affected by these extrinsic factors in either both or one of the medium-distance migrants, such effects were not found in the long-distance migrant.ConclusionsOur results suggest that the general timing of departures in songbirds is affected by the species-specific migration strategy and associated time constraints. Further, they imply that the assessment and usage of specific extrinsic factors, i.e., weather conditions, as departure cues is adjusted based on this migration strategy, with the long-distance migrants being least selective at departure. Other intrinsic and extrinsic factors, however, seem to be used as departure cues independent of migration strategy.

All aerobic organisms are to some degree affected by oxidative stress, which is an imbalance between pro-oxidants and antioxidants in favor of the former. Pro-oxidants can damage DNA, proteins and lipids, and as such oxidative stress can carry considerably fitness costs. In mammals excessive calorie intake is a known cause of oxidative stress. We investigated whether in migrant birds, which typically engage in over-eating in between flights (refueling), high food intake causes oxidative stress. In an experiment we compared levels of plasmatic total non-enzymatic antioxidant capacity (AOX) and oxidative damage (lipid peroxidation) between migrants repeatedly fasted and refed (simulating the flight-refuel cycle of wild migrants), and migrants on ad libitum food. We found that refueling increased AOX, an effect mainly attributable to an increase in uric acid level, an antioxidant that is produced during protein metabolism. Accordingly, variation in AOX was mainly explained by the refueling birds' food intake. However, food intake in migrants on ad libitum food did not explain any variation in AOX. Refueling did not affect lipid peroxidation, nor were its levels explained by food intake. We propose that over-eating migrants retain uric acid, which might be a very low cost mechanism to forego oxidative damage.

For migrating birds, late arrival at the breeding grounds reduces reproductive success. Hence, while en route, late migrants are expected to accelerate migration. Most time can be saved by shortening stopovers, periods of rest and fuel replenishment (refuelling) in between flight bouts. In several field studies late migrants have indeed been observed to shorten stopovers. The length of stopovers is assumed to depend, at least partly, on refuelling rate (fuel deposition rate, FDR). Therefore, shorter stopovers later in the spring migration season may be explained by a seasonal increase in FDR. We tested this idea by measuring both migratory restlessness (an accurate proxy for departure likelihood from stopover) and FDR in the same set of temporarily caged wheatears, Oenanthe oenanthe, a long-distance songbird migrant, throughout spring migration. We found that the amount of migratory restlessness increased with the progress of the season, indicating higher departure likelihood in late migrants. Clearly, however, late migrants in our sample did not accelerate migration by increasing their FDR; daily FDR was relatively constant during the season in lean birds, and even decreased over the season in fat birds. There was little seasonal variation in daily food intake in both lean and fat birds. Because we eliminated and/or corrected for environmental factors influencing departure likelihood, these results strongly indicate that an endogenous time programme is involved in the regulation of departure from stopover in spring.

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.

Corticosterone at baseline levels is thought to be mainly involved in the regulation of uptake, storage and release of energy, processes central to avian migration. Consequently, corticosterone levels are thought to be upregulated during migration, but the temporal pattern of its secretion during migration is not well defined. For example, although it appears that corticosterone levels decrease from flight to stopover, it is unknown if levels at stopover are still elevated and it is largely unclear how these levels compare to non-migratory life-history stages. Furthermore, what role corticosterone plays in crucial migratory processes, such as refueling and departure from stopover, is far from understood. We here determined baseline corticosterone levels in migrating and resident common blackbirds ( Turdus merula ), sampled simultaneously on Helgoland, a stopover site that also supports a sedentary breeding population. In autumn, migrants had higher corticosterone levels than residents, but in spring levels did not differ between the two groups. Corticosterone levels of migrants were very similar in spring and autumn, whereas in residents levels tended to be higher in spring than autumn. Higher levels in residents in spring than autumn most likely reflect the higher daily workload faced by birds during the pre-breeding than the post-breeding period. Our study thus indicates that, relative to the levels observed in residents in autumn, in spring baseline corticosterone levels were moderately elevated in both migrants and residents and that in autumn levels were moderately elevated in migrants only. Currently, corticosterone’s main function at stopover is thought to lie in the regulation of departure. Because most migrant blackbirds stay only one or two days on Helgoland, our results are in line with this idea and suggest that migrating blackbirds up-regulated their corticosterone level in anticipation of an oncoming flight bout.

In many migratory birds, males precede females during migration and arrival at the breeding sites. Three proximate mechanisms are proposed to explain this phenomenon of protandry: males 1) winter closer to breeding sites, 2) start spring migration earlier, and/or 3) migrate faster than females. So far, the relative contribution of these mechanisms to protandry is unknown. The present study investigated the importance of each of the 3 proximate mechanisms of protandry for a songbird migrant wintering in Africa, the northern wheatear (Oenanthe oenanthe). Two subspecies co-occur in Europe on migration, of which the leucorhoa northern wheatears breeding from Iceland to Canada have to cross the North Atlantic, whereas the nominate form breeding in Europe does not face any significant sea barrier. We show that the leucorhoa subspecies had a significantly higher degree of protandry at stopover sites across Europe than the oenanthe subspecies (−6 vs. −2 days). Leucorhoa northern wheatear’s higher degree of protandry was associated with a larger age effect, in which old males preceded young males, and greater sex-specific differences in wing shape and refueling yielding higher migration speeds in males than females. In oenanthe northern wheatears, light-level geolocators revealed that males did not winter closer to the breeding sites or migrate faster than females, but initiated spring migration earlier. Our results demonstrate that the significance of the mechanisms causing protandry can differ between related taxa and highlight the importance of the advancement in male arrival date with age as a potential factor shaping the degree of protandry.

A widespread phenomenon in migrant birds is that they travel faster in spring than in autumn. During migration birds spend most time at stopover sites and, correspondingly, the faster spring migration is mainly explained by shorter stopovers in spring than autumn. Because a main purpose of stopovers is to replenish the fuel used in flight, a higher rate of fuel deposition (FDR) in spring is thought to explain the shorter stopovers and hence shorter total duration of migration in spring. Critical migratory processes, including the onset and extent of pre-migratory fueling, are endogenously regulated. It is therefore not unlikely that refueling at stopover sites is, at least partly, also under endogenous control. We here tested whether there is an endogenous seasonal difference in food intake and FDR, which could contribute to shorter stopovers and hence faster migration in spring. We measured daily food intake and daily FDR in two subspecies of the northern wheatear Oenanthe oenanthe, temporarily confined at stopover under identical constant indoor conditions in spring and autumn. The two wheatear subspecies differed markedly in absolute food intake and FDR. Within subspecies, however, food intake and FDR did not differ between spring and autumn, indicating that faster spring migration in northern wheatears is not explained by an endogenously controlled seasonal difference in birds’ motivation to refuel. To further substantiate this claim, similar measurements should be taken at other locations along northern wheatears’ migration routes. Comparable experiments in other species could test the generality of our results.

Numerous studies in both the field and laboratory illustrate factors involved in the endogenous, intrinsic and environmental control of avian migration, but we are lacking an integrated individual-based approach, connecting field and laboratory studies in a single species. The Northern Wheatear Oenanthe oenanthe appeared very feasible for an integrated individual-based one-species approach in the study of the control of bird migration as it can be studied individually in the field as well as kept indoor under controlled ‘common-garden’ conditions. We here present a brief overview of the results collected in our wheatear studies on the intrinsic disposition and extrinsic factors that control migratory behavior in an obligate migratory species.

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.

Cooperative breeding – in which some sexually mature individuals forgo independent breeding, join a group as subordinate and help to raise the offspring of others – occurs in at least 3mammals) and 9birds) of vertebrates. Because helping others is costly, this behaviour contradicts the concept of ‘selfish’ natural selection. The intriguing evolutionary paradox of such seemingly altruistic behaviour has, therefore, been the focus of much study aiming to unravelling the evolutionary drivers underlying cooperative breeding. The benefits of group living, costs of dispersal and constraints of limited available independent breeding positions may persuade individuals to delay independent breeding and remain as subordinates within a group. However, it is the range of subsequent benefits (indirect benefits – such as improving reproduction and survival of related individuals or direct benefits – such as gaining breeding experience, benefits of future cooperation with raised recruits or gaining a share in reproduction) that favour the evolution of helping. Key Concepts The evolution of cooperative breeding is best understood by considering the benefits for helpers obtained from delayed dispersal and from helping, and the benefits helpers impose on breeders. Subordinate individuals are expected to stay when there are constraints on independent breeding and/or when they obtain benefits in their resident territory. Subordinates individuals can help related individuals and gain indirect (kin selected) fitness benefits, but in many species, helpers can also be unrelated; direct benefits of helping (like parentage, being allowed to stay) may thus be more important than commonly assumed. Breeders can benefit from helpers, either because these improve their reproductive success or because breeders can reduce their own investment in the offspring. The surplus of reproductive capable individuals and the effects of helpers on survival and reproduction can have important implications for population dynamics in cooperatively breeding species.

Most avian migrants alternate flight bouts, characterized by high metabolic rates, with stopovers, periods of fuel replenishment through hyperphagia. High-energy metabolism and excessive calorie intake shift the balance between damaging prooxidants and antioxidants toward the former. Hence, migration likely affects the oxidative balance of birds. Migratory flight indeed appears to cause oxidative damage; however, whether migration affects the oxidative state of birds at stopover is unclear. Therefore, we compared total nonenzymatic antioxidant capacity (AOX) and malondialdehyde concentration (MDA; a measure of lipid peroxidation) in the plasma of migrant and resident common blackbirds. We also determined plasmatic uric acid (UA) and fatty acid (FA) concentrations and calculated a FA peroxidation index. Birds were sampled during autumn migration at a stopover site that also supports a sedentary blackbird population. Migrants had higher AOX than residents, also after correcting for UA concentration. Migrants tended to have higher FA peroxidation indexes than residents, indicating that the energy source of migrants contains higher concentrations of peroxidizable FAs. However, the two groups did not differ in MDA concentration, also not after correcting for peroxidation index. Peroxidation-corrected MDA concentration was negatively correlated with UA-corrected AOX. In other words, individuals with low nonenzymatic AOX suffered more from lipid peroxidation than individuals with high nonenzymatic AOX. These results together indicate that migrant blackbirds invest in antioxidant defenses to reduce oxidative damage to lipids, likely representing an adaptation to diminish the physiological costs of migration.

A variety of methods have been used to study the relationship between wind conditions and departure decisions of migrant birds at stopover sites. These methods are either costly or suffer from inaccuracy in determining whether or not an individual has resumed migration. Here we present a novel and simple approach to studying the relationship between wind conditions and departure likelihood. Northern Wheatears Oenanthe oenanthe caught during stopover were temporarily caged to measure their nocturnal migratory restlessness, which is an accurate proxy for their individual departure likelihood. We then related the degree of nocturnal restlessness to wind conditions prevailing at the time of capture. Confirming the general pattern from previous studies of departure, the intensity of nocturnal migratory restlessness, and hence departure likelihood, increased with increasing wind support towards the migratory goal. This suggests that approximating the propensity to depart by measuring nocturnal migratory restlessness is a reliable way to study the effect that wind conditions experienced during stopover has on the departure decision of migrants. Our study also shows that nocturnal migrants possess the ability to use information gathered during the day for their departure decisions at night. Because measuring migratory restlessness is straightforward and inexpensive, our approach is ideally suited to test hypotheses regarding spatio-temporal variation in wind selectivity in migrating birds.

Most migrating birds alternate flight bouts with stopovers, during which they rest and replenish the fuel used during flight (refueling). The rate of refueling (fuel deposition rate, FDR) affects stopover duration, and hence is an important determinant of the overall time required for migration. Although environmental and endogenous factors affect FDR, the urge to refuel depends on the anticipated distance to be travelled and possibly also on the amount of fuel used during the flight preceding stopover. Combining a field study with a fasting–refueling experiment on long-term captive songbirds, we tested whether the extent of fuel loss prior to refueling indeed affects FDR. In the field study, we took a comparative approach and determined FDR in two subspecies of northern wheatear (Oenanthe oenanthe) that differ greatly in the distance flown, and thus the extent of fuel used to reach our study (stopover) site. As both winter in western sub-Sahelian Africa, they face the same remaining migration distance. We found that FDR was higher in the subspecies that uses more fuel to get to our study site. Solidifying this result, in the experiment on captive northern wheatears, we found that the extent of fuel loss as a consequence of fasting explained most of the variation in subsequent FDR. The observation that experimental birds losing little fuel did not maximize their FDR suggests there are costs to rapid refueling. Our study shows that FDR is shaped not just by current environmental and endogenous conditions but also by fuel loss prior to refueling.

Field endocrine studies providing new comparisons for inference into the evolutionary and ecological factors shaping organismal physiology are important, often yielding novel physiological insights. Here, we explored factors associated with the sex steroid hormone concentrations and adrenocortical response to capture stress in Hog Island boas (Boa constrictor imperator) in the Cayos Cochinos archipelago of Honduras to generate comparative field hormone data from a tropical reptile and test the island tameness hypothesis. Baseline concentrations of testosterone, corticosterone, estradiol, and progesterone were measured during the wet and dry seasons, and an acute stressor of 1 h in a cloth bag was used to assess the stress response. Plasma steroid concentrations in these snakes were generally low in comparison to other taxa. Higher testosterone concentrations in males and higher estradiol and corticosterone concentrations in females were observed during the wet season compared to the dry season, which may be indicative of mating activities and vitellogenesis during this period. Snakes displayed a 15-fold increase in corticosterone concentrations in response to capture stress, a rise that was not impacted by whether a snake had been captured during previous years. The adrenocortical stress response was greater in males and positively related to body temperature. We suggest that this system merits future inquiries into the physiology and behavior of B. c. imperator, particularly as a model for studying insular impacts on diverse life history characters.

Migrating birds spend most of their time at stopover sites where they replenish the fuel used during flight, termed refueling. The overall time of migration thus largely depends on the duration of stopovers, and factors shaping stopover duration therefore are of interest. A handful of field studies have shown that the likelihood of departure from stopover sites increases with poor feeding conditions. However, food availability and stopover duration are generally difficult to quantify accurately in the field. Results of fasting-refueling experiments on captive birds using migratory restlessness (Zugunruhe) as a proxy for departure likelihood are mixed. Although Zugunruhe usually decreased with refueling, fasting often failed to increase Zugunruhe. In addition, some experiments lacked randomization. In a fasting-refueling experiment on Northern Wheatears (Oenanthe oenanthe), using birds as their own control in a randomized fashion, we found that fasting increased Zugunruhe, whereas refueling decreased Zugunruhe. These results show that the motivation to migrate, quantified by Zugunruhe, is affected by changes in food availability. Furthermore, Zugunruhe during refueling did not depend on fuel reserves left after fasting, but tended to decrease with the amount of fuel lost during fasting. We discuss why extent of fuel loss may be a better predictor of stopover duration than fuel reserves.

Migratory birds rely on fueling prior to migratory flights. Fueling in migrants is controlled by intrinsic as well as extrinsic factors. From captive studies we have started understanding the internal mechanisms controlling bird migration. Field studies have demonstrated the effects of external factors, such as food availability, weather, competitors, parasites or diseases, on the stopover behavior of migrants. However, an integrated approach is still missing to study coherently how the innate migration program interacts with the varying environmental cues and to estimate the contribution of the innate migration program and the environment to realized migration. The northern wheatear Oenanthe oenanthe offers a unique opportunity for integrated studies. It breeds across almost the whole Holarctic with just a “gap” between eastern Canada and Alaska. All breeding populations overwinter in sub-Saharan Africa which makes the northern wheatear one of the most long-distant migratory songbirds with extraordinary long non-stop flights across oceans. It is a nocturnal migrant which travels without parental or social aid/guidance. Thus, young birds rely entirely on endogenous mechanisms of timing, route selection and fueling on their first outbound migration. By establishing indoor housing under controlled conditions the endogenous control mechanisms of northern wheatear migration could be revealed. At the same time, environmental factors controlling fueling could be investigated in the field. On migration wheatears occur in a variety of habitats with sparse vegetation where their stopover behavior could be quantitatively studied in the light of “optimal migration” theory by the use of remote balances, radio-tagging and even experimentally manipulated food availability. The present paper summarizes our approach to understand the control of migration in northern wheatears by combining field and laboratory studies at various spatial and temporal scales, and linking various sub-disciplines.

Corticosterone, at baseline and moderately elevated levels, is thought to regulate energy mobilization during the predictable life-history cycle. In birds, corticosterone is known to be moderately elevated during migration, and some experiments on captive, but migratory active birds have shown that exogenous corticosterone can positively affect food intake and fat deposition, i.e. fueling. We present observations which indicate that in wild birds endogenous corticosterone does not promote refueling during migratory stopovers. We took a comparative approach and studied two subspecies of Northern wheatears (Oenanthe oenanthe) during their simultaneous spring stopovers on Helgoland, a small island some 50km off the German coast. In spring O. oenanthe have to travel relatively short distances from Helgoland to their next stopover or breeding sites, whereas Oenanthe leucorhoa face a lengthy overseas journey. Consequently, for their next flight bout leucorhoa wheatears deposit more fuel, more rapidly than oenanthe wheatears. Corticosterone levels, however, were lower in leucorhoa than oenanthe wheatears, contradicting the idea that corticosterone promotes migratory refueling. This finding was solidified by the observation that actual fuel deposition rate was negatively correlated with corticosterone level. We also observed a positive correlation between corticosterone level and fuel stores. Together these findings suggest that, rather than promoting migratory refueling, corticosterone may function as a readiness cue, with levels increasing towards departure from the stopover site.

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.

We describe the reproductive cycle of Northern Pacific rattlesnakes (Crotalus oreganus) by quantifying steroid hormone concentrations and observing reproductive behaviors in free-ranging individuals. Additionally, we examined reproductive tissues from museum specimens. Plasma steroid hormone concentrations were quantified for both male and female snakes throughout the active season (March–October). We measured testosterone (T), 5a-dihydrotestosterone (DHT), and corticosterone (B) concentrations in both sexes and 17b-estradiol (E2) and progesterone (P) in females only. We observed reproductive behaviors (e.g., consortship, courtship, and copulation) in the field and measured testis and follicle size in male and female snakes from museum collections to relate steroid hormone concentrations to the timing of reproductive events. Our study revealed that C. oreganus in central California exhibits a bimodal pattern of breeding, with most mating behavior occurring in the spring and some incidences of mating behavior observed in late summer/fall. Each breeding period corresponded with elevated androgen (T or DHT) levels in males. Testes were regressed in the spring when the majority of reproductive behavior was observed in this population, and they reached peak volume in August and September during spermatogenesis. Although we did not detect seasonal variation in female hormone concentrations, some females had high E2 in the spring and fall, coincident with mating and with increased follicle size (indicating vitellogenesis) in museum specimens. Females with high E2 concentrations also had high T and DHT concentrations. Corticosterone concentrations in males and females were not related either to time of year or to concentrations of any other hormones quantified. Progesterone concentrations in females also did not vary seasonally, but this likely reflected sampling bias as females tended to be underground, and thus unobtainable, in summer months when P would be expected to be elevated during gestation. In females, P was positively correlated with T and DHT, and E2 was positively correlated with T.

Latitudinal variation in life-history traits has been the focus of numerous investigations, but underlying hormonal mech- anisms have received much less attention. Steroid hormones play a central role in vertebrate reproduction and may be associated with life-history trade-offs. Consequently, circulating concentrations of these hormones vary tremendously across vertebrates, yet interspe- cific geographic variation in male hormone concentrations has been studied in detail only in birds. We here report on such variation in amphibians and reptiles, confirming patterns observed in birds. Us- ing phylogenetic comparative analyses, we found that in amphibians, but not in reptiles, testosterone and baseline corticosterone were positively related to latitude. Baseline corticosterone was negatively related to elevation in amphibians but not in reptiles. For both groups, testosterone concentrations were negatively related to breed- ing-season length. In addition, testosterone concentrations were pos- itively correlated with baseline corticosterone in both groups. Our findings may best be explained by the hypothesis that shorter breed- ing seasons increase male-male competition, which may favor in- creased testosterone concentrations that modulate secondary sexual traits. Elevated energetic demands resulting from greater reproductive intensity may require higher baseline corticosterone. Thus, the pos- itive relationship between testosterone and corticosterone in both groups suggests an energetic demand for testosterone-regulated be- havior that is met with increased baseline glucocorticoid con

Knowing why certain individuals refrain from reproduction is fundamental to our understanding of the evolution of cooperative breeding. Although many studies have focused on the role of life history and ecological factors, the proximate mechanisms underlying this behaviour have received considerably less attention. We determined the natural variation in plasma testosterone (T) level and cloacal protuberance (CP) size - an indication of sperm storage - in successfully reproducing primary male and in reproductively inactive subordinate male Seychelles warblers (Acrocephalus sechellensis). Primary males had significantly higher T levels during female's fertile period and significantly larger CPs throughout the breeding season than subordinate males. To test if the lower T levels of subordinates were caused by their social status per se, we experimentally promoted subordinates to primary males, by permanent removal of primary males from their territories. The experimentally promoted males increased T production and CP sizes significantly, suggesting former suppression by primary males. However, T levels of the promoted males only marginally increased. As this could not be explained by their younger age, we conclude that recovery from social suppression may take a substantial time period.

In many species, territory ownership is a prerequisite for reproduction; consequently, factors that affect success in territory acquisition can have a large impact on fitness. When competing for territories, some individuals may have an advantage if, for example, they are phenotypically superior or more familiar with the site than others. The relative importance of the many factors involved in territory acquisition is, at present, unclear. We studied patterns of natural territory acquisition in a closed and saturated population of Seychelles warblers. Furthermore, by removing breeders, we experimentally investigated the relative importance, to territory acquisition, of a range of factors and assessed whether this differed between the sexes. In both sexes, the main route to natural territory acquisition was to disperse from the natal territory to immediately claim a vacant dominant position. Males were older than females when acquiring a territory for the first time. In the removal experiment, for both sexes, the proximity of an individual's natal territory to a vacant dominant position was positively related to the individual's chance of claiming the vacancy. Older males were more likely to gain an experimental vacant dominant position than were young males, whereas age did not affect territory acquisition in females. In the Seychelles warbler, the degree of intrasexual competition for territory ownership may be stronger for males than for females because territory ownership is a prerequisite for male reproduction, whereas females can reproduce on their natal territory. In such competition, young males subsequently lose out to older ones. Copyright 2009, Oxford University Press.

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.

The presence of parents in the natal territory may play an important, but often overlooked, role in natal dispersal and the consequent acquisition of a territory. Living with parents in a territory may confer a fitness advantage to subordinates through, for example, the nepotistic behavior of the parents or indirect benefits gained by helping to raise nondescendent kin. When a parent is replaced by a stepparent, such advantages are reduced or disappear and, as a result, subordinates may disperse. Subordinates that disperse after parent replacement may be constrained in their timing of dispersal, which could have negative fitness consequences. In the cooperatively breeding Seychelles warbler, we show that when a parent was naturally replaced or experimentally removed and subsequently replaced by a stepparent from outside the territory, subordinates were more likely to disperse than when both parents remained in the natal territory. Furthermore, subordinates dispersing from territories in which one or both parents had been replaced were less likely to acquire a breeder position than subordinates dispersing when both parents were still on the natal territory. Our findings suggest that the presence of parents in the natal territory may promote delayed dispersal and facilitate the eventual acquisition of a breeder position outside the natal territory. Our results support the idea that the prolonged parental care, which long-lived species are able to provide, may have selected for family living. Copyright 2007, Oxford University Press.

1. Variation in survival, a major determinant of fitness, may be caused by individual or environmental characteristics. Furthermore, interactions between individuals may influence survival through the negative feedback effects of density dependence. Compared to species in temperate regions, we have little knowledge about population processes and variation in fitness in tropical bird species.2. To investigate whether variation in survival could be explained by population size or climatic variables we used capture-recapture models in conjunction with a long-term data set from an island population of the territorial, cooperatively breeding Seychelles warbler (Acrocephalus sechellensis). The lack of migration out of the study population means that our results are not confounded by dispersal.3. Annual survival was high, both for adults (84%) and juveniles (61%), and did not differ between the sexes. Although there was significant variation in survival between years, this variation could not be explained by overall population size or weather variables.4. For territorial species, resource competition will work mainly on a local scale. The size of a territory and number of individuals living in it will therefore be a more appropriate measure of density than overall population density. Consequently, both an index of territory quality per individual (food availability) and local density, measured as group size, were included as individual covariates in our analyses.5. Local density had a negative effect on survival; birds living in larger groups had lower survival probabilities than those living in small groups. Food availability did not affect survival.6. Our study shows that, in a territorial species, although density-dependent effects might not be detectable at the population level they can be detected at the individual territory level - the scale at which individuals compete. These results will help to provide a better understanding of the small-scale processes involved in the dynamics of a population in general, but in particular in tropical species living in relatively stable environments.

Th e latitudinal increase in extra-pair paternity (EPP) rates in birds suggests broad selective benefi ts to low EPP rates in the tropics. However, we have few EPP data from tropical birds, particularly from species with close relatives at high latitudes. Here, we report EPP rates in two resident equatorial populations of rufous-collared sparrow Zonotrichia capensis, a genus well-represented at high latitudes. We found 64% and 60% of broods contained extra-pair off spring, and 42% and 52% of all young were extra-pair. EPP rates were similar in these populations, despite clear diff erences in elevation, temperature, rainfall, and breeding season length. Th ese fi ndings provide evidence that EPP rates in tropical birds can be as high as those observed in temperate birds, and suggest that the selective pressures acting on EPP rates vary markedly across tropical birds.

In captivity, migratory birds show increased activity during the time that they would normally migrate. The phenology and intensity of such ‘migratory restlessness’ has been shown to mirror species- and population-specific migration patterns observed in the wild and has consequently been used as a proxy for the motivation to migrate. Many studies doing so, however, were aiming to explain among-individual variation in migratory behaviour or traits, and not species- or population-specific traits. These studies thus assumed that, also at the level of the individual, migratory restlessness is an accurate proxy for the motivation to migrate. We tested this assumption for the first time and found that it holds; individuals showing very little migratory restlessness remained at stopover for longer than one night, whereas most individuals showing more restlessness departed sooner. This finding validates the use of migratory restlessness as a proxy for the motivation to migrate, thereby justifying the conclusions made in a large body of research on avian migration.

The decision made by migrating birds to stop refueling and to depart from stopover depends on cues from innate rhythms, intrinsic factors such as fuel reserves, and extrinsic factors such as weather conditions. The physiological mechanism behind this decision, however, is largely unexplored. The transition from refueling to flight involves an increase in both locomotion and energetic demands. Because, at baseline levels, corticosterone stimulates locomotion and is involved in the mobilization of energy, this hormone could encourage departure of migrants. We collected field data on baseline corticosterone, migratory restlessness, and actual departure in northern wheatears at stopover. Additionally, in refueling long-term captive conspecifics, we measured migratory restlessness while simultaneously collecting droppings to determine glucocorticoid metabolite (GCM) levels. We found that migratory restlessness at stopover was positively correlated with corticosterone level. Similarly, in refueling long-term captive birds, migratory restlessness was positively correlated with concurrently measured GCM levels in droppings. To our best knowledge, our study is the first to simultaneously measure a hormonal signal and migratory restlessness. In conclusion, our results are consistent with the hypothesis that, by increasing locomotor activity, baseline corticosterone is involved in the regulation of departure of migrants at stopover. Future studies could reveal how corticosterone is up-regulated in migrants that are ready to depart.

We tested the hypothesis that mother birds counterbalance the negative effects of hatching asynchrony for later–hatched chicks by increasing the yolk androgen concentrations in consecutive eggs of their clutch. In doing so, they may adaptively tune each offspring9s competitive ability and, thus, growth and survival. However, evidence in support of this hypothesis is contradictory. The yolk concentrations of maternal androgens in the eggs of black–headed gulls increase significantly with the laying order of the eggs in a clutch. We experimentally tested the functional consequences of this increase on chick development under natural conditions by injecting eggs with either an oil or androgen solution. We created experimental clutches in which androgen levels either stayed constant or increased with laying order while controlling for differences in egg quality by using only first–laid eggs. We then compared development, growth and survival between these broods. Androgen treatment enhanced embryonic development because androgen–treated eggs hatched half a day earlier than controls, while their size at hatching was similar to oil–treated controls. Androgen treatment did not increase chick survival, but it enhanced growth. Androgen–treated, third–hatched chicks had a higher body mass and longer legs than thirdndash;hatched chicks that hatched from oil–treated eggs. At the same time, growth of first chicks (which were all oil treated) was reduced by the presence of two androgen–treated siblings, suggesting that yolk androgens enhance the competitive ability of later–hatched chicks. Our results support the hypothesis that transfer of different amounts of androgens to the eggs of a clutch is a mechanism by which mothers maximize their reproductive output.

Mating systems can vary among species and populations and thus influence evolutionary trajectories, ecological traits, and population demography. The siring of offspring by an extrapair male, or extrapair paternity (EPP), is a widespread and varied phenomenon in all vertebrate classes. However, we do not understand all of the factors associated with variation in EPP rates. The breeding synchrony hypothesis suggests that EPP rates should increase with latitude and elevation, whereas the paternal care hypothesis predicts that EPP rates should decrease with elevation. To address these hypotheses, we investigated how population EPP rates vary over elevation and latitude in emberizid sparrows. In comparative analyses including the effects of phylogeny, the relationship between EPP rates and elevation depended on latitude. EPP rates were greater in higher-latitude populations. But within higher-latitude populations, EPP rates decreased with increasing elevation. These findings provide support for both the breeding synchrony and paternal care hypotheses, suggesting that in lower-latitude, higher-elevation populations, the need for male parental care does not outweigh the benefits of seeking extrapair fertilizations in populations with relatively synchronous breeding. In contrast, at higher-latitude, higher-elevation sites, the need for male parental care is greater and might drive lower rates of extrapair mating despite highly synchronous breeding.

In most monogamous bird species, circulating testosterone concentration in males is elevated around the social female's fertile period. Variation in elevated testosterone concentrations among males may have a considerable impact on fitness. For example, testosterone implants enhance behaviours important for social and extra-pair mate choice. However, little is known about the relationship between natural male testosterone concentration and sexual selection. To investigate this relationship we measured testosterone concentration and sexual signals (ventral plumage colour and tail length), and determined within and extra-pair fertilization success in male North American barn swallows (Hirundo rustica erythrogaster). Dark rusty coloured males had higher testosterone concentrations than drab males. Extra-pair paternity was common (42% and 31% of young in 2009 and 2010, respectively), but neither within- nor extra-pair fertilization success was related to male testosterone concentration. Dark rusty males were less often cuckolded, but did not have higher extra-pair or total fertilization success than drab males. Tail length did not affect within- or extra-pair fertilization success. Our findings suggest that, in North American barn swallows, male testosterone concentration does not play a significant direct role in female mate choice and sexual selection. Possibly plumage colour co-varies with a male behavioural trait, such as aggressiveness, that reduces the chance of cuckoldry. This could also explain why dark males have higher testosterone concentrations than drab males. Published version

Males may face a trade-off between caring for offspring and pursuing additional matings. In birds, the androgen testosterone has been suggested to be a key proximate mediator in this trade-off for several reasons. At the population level, high testosterone is typically associated with the period of intense male–male competition over females, whereas low testosterone is associated with the period of paternal care. In addition, males with experimentally elevated testosterone during provisioning feed their young at a lower rate than control males. Nearly all studies observing these patterns, however, ignore the tremendous variation in endogenous testosterone concentration that exists within a population of males, even during the same breeding stage. Because selection acts at the level of individual, this variation has to be taken into account when studying proximate mechanisms mediating the paternal and mating effort trade-off. Studying barn swallows we here show that, within males, testosterone concentrations were not higher around the fertile period of the social mate than during nestling feeding. More importantly, 30% of males showed no decrease in testosterone concentration between these 2 periods. Further, male feeding effort was not related to testosterone concentration during feeding. These results indicate that, at least in barn swallows, endogenous testosterone is not a key mediator in the trade-off between paternal effort and mating effort. Our results also stress that to understand how selection has shaped temporal testosterone profiles and action, it is crucial to study the relationship between testosterone and traits contributing to fitness at the level of the individual. Key words: barn swallow, endogenous testosterone, extrapair, mating effort, nestling feeding, paternal effort. [Behav Ecol]

We observed a change in the sex-specific rate of delayed natal dispersal in a stable population of Seychelles warblers over a period of 20 years. At first, females were more likely to delay dispersal in their first year of life than were males, whereas later there was no sex bias in the rate of delayed natal dispersal. Similarly, the female-bias in helping-at-the-nest and the male-bias in floating have also weakened over time. These changes may have resulted from the decrease in variation in territory quality observed in the population over the study period. Our findings strengthen the view that natal dispersal is a highly plastic response to local ecological and social circumstances, and clearly show that rates of sex-biased dispersal cannot be considered a species or population constant. Our study also highlights the importance of collecting long-term datasets to understand complex behaviour such as natal dispersal.

We describe the reproductive cycle of Northern Pacific rattlesnakes (Crotalus oreganus) by quantifying steroid hormone concentrations and observing reproductive behaviors in free-ranging individuals. Additionally, we examined reproductive tissues from museum specimens. Plasma steroid hormone concentrations were quantified for both male and female snakes throughout the active season (March-October). We measured testosterone (T), 5alpha-dihydrotestosterone (DHT), and corticosterone (B) concentrations in both sexes and 17beta-estradiol (E2) and progesterone (P) in females only. We observed reproductive behaviors (e.g., consortship, courtship, and copulation) in the field and measured testis and follicle size in male and female snakes from museum collections to relate steroid hormone concentrations to the timing of reproductive events. Our study revealed that C. oreganus in central California exhibits a bimodal pattern of breeding, with most mating behavior occurring in the spring and some incidences of mating behavior observed in late summer/fall. Each breeding period corresponded with elevated androgen (T or DHT) levels in males. Testes were regressed in the spring when the majority of reproductive behavior was observed in this population, and they reached peak volume in August and September during spermatogenesis. Although we did not detect seasonal variation in female hormone concentrations, some females had high E2 in the spring and fall, coincident with mating and with increased follicle size (indicating vitellogenesis) in museum specimens. Females with high E2 concentrations also had high T and DHT concentrations. Corticosterone concentrations in males and females were not related either to time of year or to concentrations of any other hormones quantified. Progesterone concentrations in females also did not vary seasonally, but this likely reflected sampling bias as females tended to be underground, and thus unobtainable, in summer months when P would be expected to be elevated during gestation. In females, P was positively correlated with T and DHT, and E2 was positively correlated with T.

Cooperative breeding – in which some adults forgo independent breeding and remain as subordinates within a group helping to raise the offspring of others – occurs in between 3% and 10% of vertebrates. The structure of such systems varies greatly, from pairs with helpers-at-the-nest to communal breeders, andmayinclude young helpers or post-reproductive ‘grandparents’. That some individuals spend part, or all, of their lives helping others to reproduce contradicts the concept of ‘selfish’ natural selection and provides an intriguing evolutionary paradox. When and why such apparently altruistic behaviour occurs has, therefore, been the focus of much study. Although constraints and the benefits of group living, may persuade individuals to remain as subordinates, indirect ‘kin’ benefits or direct benefits (such as becoming a breeding helper) appear to favour the subsequent evolution of helping. Cooperative behaviour can have wide-ranging consequences, not only on the biology of the cooperative species, but also on those species it interacts with.

In this study, we test whether patterns of territory inheritance, social mate choice and female-biased natal dispersal act as inbreeding avoidance mechanisms in the cooperatively breeding Seychelles warbler. Our results show that Seychelles warblers do not reduce the likelihood of inbreeding by avoiding related individuals as mates. The occurrence of natural and experimentally induced territory inheritance did not depend on whether the remaining breeder was a parent of the potential inheritor or an unrelated breeder. Furthermore, dispersing individuals were no less related to their eventual mates than expected given the pool of candidates they could mate with. The female bias in natal dispersal distance observed in the Seychelles warbler does not facilitate inbreeding avoidance because, contrary to our prediction, there was no sex difference in the clustering of related opposite sex breeders around the natal territories of dispersers. As a result, the chance of females mating with relatives was not reduced by their greater dispersal distance compared with that of males.