Stress,immunocompetence and leukocyte profiles of pied flycatchers in relation to brood size manipulation

The two main trade-offs considered determining reproductive patterns in iteroparous organisms are the one between current and future reproduction, and the one between the number and quality of offspring. Recently, it has been suggested that these trade-offs may be mediated by stress-induced reduction in immunocompetence. To test the hypothesis that stress reduces immune function, we investigated the effects of brood size manipulation on stress hormone levels, leukocyte profiles and immune responses against challenge with novel antigens in nestling and parent male pied flycatchers ( Ficedula hypoleuca). In male parents, heterophil (H) and lymphocyte (L) numbers, as well as H/L ratio increased with experimentally enlarged brood size, and corticosterone levels tended to do so, indicating that high parental work load altered their stress level and physiological state. Despite this, we found no effects on humoral immune responsiveness, measured as antibody production against diphtheria-tetanus vaccine. In nestlings, heterophil numbers and H/L ratio increased in enlarged broods, whereas T-cell-mediated immune responsiveness, measured against phytohemagglutinin (PHA), decreased in enlarged broods. The results support the view that growth-stress-induced immunosuppression may be an important physiological pathway mediating the trade-off between the number and viability of offspring. The difference in the observed immune-related responses between nestlings and males may be because we measured different aspects of the immune system (cellular vs humoral). However, it may also be a result of males lowering their own costs by feeding less, (and their mate possibly compensate by feeding more), whereas nestlings cannot escape the costs of increased intra-brood competition.     

Nestling cuckoos,Cuculus canorus,exploit hosts with begging calls that mimic a brood

Nestling cuckoos, Cuculus canorus, eject host eggs or young from the nest and are then raised alone by the hosts. Using reed warblers, Acrocephalus scirpaceus, as hosts, we investigated how the single cuckoo chick can command the same provisioning rate as a whole brood of host young. Large size alone is not sufficient to stimulate adequate provisioning because single blackbird, Turdus merula, or song thrush, T. philomelos, chicks of the same mass as a cuckoo were fed at a lower rate. Our experiments show that the key stimulus is the cuckoo chick''s rapid begging call (''si, si, si, si ...''), which sounds remarkably like a whole brood of host chicks, and which it matched in calling rate. When single blackbird or song thrush chicks were accompanied by loudspeakers that broadcast either cuckoo begging calls or calls of a brood of reed warblers, the hosts increased their provisioning rate to that for a cuckoo chick. We suggest that the cuckoo needs vocal trickery to stimulate adequate care to compensate for the fact that it presents a visual stimulus of just one gape.     

Host and brood parasite coevolutionary interactions covary with comparative patterns of the avian visual system

In coevolutionary arms-races, reciprocal ecological interactions and their fitness impacts shape the course of phenotypic evolution. The classic example of avian host–brood parasite interactions selects for host recognition and rejection of increasingly mimetic foreign eggs. An essential component of perceptual mimicry is that parasitic eggs escape detection by host sensory systems, yet there is no direct evidence that the avian visual system covaries with parasitic egg recognition or mimicry. Here, we used eye size measurements collected from preserved museum specimens as a metric of the avian visual system for species involved in host–brood parasite interactions. We discovered that (i) hosts had smaller eyes compared with non-hosts, (ii) parasites had larger eyes compared with hosts before but not after phylogenetic corrections, perhaps owing to the limited number of independent evolutionary origins of obligate brood parasitism, (iii) egg rejection in hosts with non-mimetic parasitic eggs positively correlated with eye size, and (iv) eye size was positively associated with increased avian-perceived host–parasite eggshell similarity. These results imply that both host-use by parasites and anti-parasitic responses by hosts covary with a metric of the visual system across relevant bird species, providing comparative evidence for coevolutionary patterns of host and brood parasite sensory systems.     

Warning signal properties covary with toxicity but not testosterone or aggregate carotenoids in a poison frog

Aposematic (warning) coloration is a highly conspicuous trait that is found throughout the animal kingdom. In several aposematic species, warning signals have been co-opted for use in conspecific communication systems; for example, in the toxic and bright orange Solarte population of the strawberry poison frog (Oophaga [Dendrobates] pumilio), the brightness of male warning coloration serves as a sexual signal by both attracting females and repelling rivals. Here, we investigate correlations between bright male warning coloration and several physiological characteristics (e.g., circulating testosterone and carotenoids and noxious alkaloids in the skin), to gain insights into the mechanisms underlying the signal variation in this population and to inform hypotheses regarding the evolutionary stability of this trait. We find that although measures of male brightness (viewer-dependent or viewer-independent) do not correlate with two classic correlates of sexually selected traits (circulating testosterone and aggregate carotenoids in the skin), male reflectance does show a positive correlation with concentrations of two xanthophyll carotenoids. Total reflectance (a viewer-independent measure of male brightness) also shows a negative relationship with aggregate pumiliotoxin in the skin, which is considered one of the major classes of defensive alkaloids in this species. Because the alkaloids used in this species’ chemical defense are acquired from dietary sources, the magnitude of the reflectance intensity of a male’s warning signal can potentially provide viewers with reliable information regarding territory quality, health, and/or current condition.     

Sperm traits negatively covary with size and asymmetry of a secondary sexual trait in a freshwater crayfish

In species where females mate promiscuously, the reproductive success of males depends both on their ability to acquire mates (pre-copulatory sexual selection) and ability of their ejaculates to outcompete those of other males (post-copulatory sexual selection). Sperm competition theory predicts a negative relationship between investment in body traits favouring mate acquisition (secondary sexual characters, SSCs) and investment in ejaculate size or quality, due to the inherent costs of sperm production. In contrast, the phenotype-linked fertility hypothesis posits that male fertilizing efficiency is reliably reflected by the phenotypic expression of male SSCs, allowing females to obtain direct benefits by selecting more ornamented males as copulation partners. In this study, we investigated the relationships between male SSCs and size and quality (viability and longevity) of ejaculates allocated to females in mating trials of the freshwater crayfish Austropotamobius italicus. We showed that the relative size of male weapons, the chelae, was negatively related to ejaculate size, and that chelae asymmetry, resulting from regeneration of lost chelipeds, negatively covaried with sperm longevity. Moreover, males allocated more viable sperm to mates from their own rather than different stream of origin. Our findings thus suggest that, according to sperm competition theory, pre-copulatory sexual selection for large weapons used in male fighting may counteract post-copulatory sperm competition in this crayfish species, and that investment in cheliped regeneration may impair ejaculate quality.     

Life history traits associated with body size covary along a latitudinal gradient in a generalist grasshopper

Animal body size often varies systematically along latitudinal gradients, where individuals are either larger or smaller with varying season length. This study examines ecotypic responses by the generalist grasshopper Melanoplus femurrubrum (Orthoptera: Acrididae) in body size and covarying, physiologically based life history traits along a latitudinal gradient with respect to seasonality and energetics. The latitudinal compensation hypothesis predicts that smaller body size occurs in colder sites when populations must compensate for time constraints due to short seasons. Shorter season length requires faster developmental and growth rates to complete life cycles in one season. Using a common garden experimental design under laboratory conditions, we examined how grasshopper body size, consumption, developmental time, growth rate and metabolism varied among populations collected along an extended latitudinal gradient. When reared at the same temperature in the lab, individuals from northern populations were smaller, developed more rapidly, and showed higher growth rates, as expected for adaptations to shorter and generally cooler growing seasons. Temperature-dependent, whole organism metabolic rate scaled positively with body size and was lower at northern sites, but mass-specific standard metabolic rate did not differ among sites. Total food consumption varied positively with body size, but northern populations exhibited a higher mass-specific consumption rate. Overall, compensatory life history responses corresponded with key predictions of the latitudinal compensation hypothesis in response to season length.     

Female plumage colour influences seasonal oxidative damage and testosterone profiles in a songbird

Across diverse taxa, morphological traits mediate social interactions and mate selection. Physiological constraints on signal elaboration have been widely documented, but the potential for trait display to influence physiological state remains poorly understood. We tested for the presence of causal links between ventral plumage colour—a trait known to covary with reproductive performance—and physiological measures in female North American barn swallows, Hirundo rustica erythrogaster. Naturally darker swallows have lower levels of plasma oxidative damage. Females manipulated to display darker ventral plumage during reproduction rapidly decreased oxidative damage, adopting the physiological state of naturally darker individuals. These results support the presence of a social mechanism that links static plumage traits with the physiological state of their bearer during trait advertisement, long after the completion of signal development.     

Parental response to brood size in a cichlid fish

This paper reports on an experiment designed to investigate whether guarding females of the cichlid Aequidens coeruleopunctatus adjust their parental behaviour in response to changes in brood size. It was found that females were easier to scare away from their broods when threatened, and stayed away for longer, after some of their fry had been removed, and that the reverse occurred when broods were experimentally augmented with foreign fry. Brood size manipulations change the expected benefits of future investment. These results therefore indicate that parental care strategies in A. coeruleopunctatus reflect future prospects for reproductive success rather than past investment.     

Effects of testosterone and corticosterone on immunocompetence in the zebra finch

The original immunocompetence handicap hypothesis (ICHH) suggested that testosterone has a handicapping effect in males by both promoting the development of sexual signals and suppressing immune function. A modified version, the stress-linked ICHH, has recently proposed that testosterone is immunosuppressive indirectly by increasing production of corticosterone. To test both the original and stress-mediated versions of the ICHH, we implanted male zebra finches taken from lines selected for divergent maximum stress-induced levels of corticosterone (high, low and control) with either empty or testosterone-filled implants. Their humoral and cell-mediated immune responses were then assessed by challenge with diphtheria:tetanus vaccine and phytohemagglutinin respectively. We found no effect of the hormone manipulations on either PHA or tetanus antibody responses, but found a significant interaction between titers of both testosterone and corticosterone on diphtheria secondary antibody response; antibody response was greatest in individuals with high levels of both hormones. There was also a significant interactive effect between testosterone treatment group and corticosterone titer on body mass; the body mass of males in the elevated testosterone treatment group decreased with increasing corticosterone titer. These results suggest that, contrary to the assumption of the stress-mediated version of the ICHH, high plasma levels of corticosterone are not immunosuppressive, but are in fact immuno-enhancing in the presence of high levels of plasma testosterone. Equally, the central assumption of the ICHH that testosterone is obligately immunosuppressive is also not supported. The same individuals with the highest levels of both hormones and consequently the most robust antibody response also possessed the lowest body mass.     

Estimating repertoire size in a songbird: a comparison of three techniques

Many animals produce multiple types of breeding vocalizations that, together, constitute a vocal repertoire. In some species, the size of an individual’s repertoire is important because it correlates with brain size, territory size or social behaviour. Quantifying repertoire size is challenging because the long recordings needed to sample a repertoire comprehensively are difficult to obtain and analyse. The most basic quantification technique is simple enumeration, where one counts unique vocalization types until no new types are detected. Alternative techniques estimate repertoire size from subsamples, but these techniques are useful only if they are accurate. Using 12 years of acoustic data from a population of rufous-and-white wrens in Costa Rica, we used simple enumeration to measure the repertoire size for 40 males. We then compared these to the estimates generated by three estimation techniques: curve fitting, capture–recapture and a new technique based on the coupon collector’s problem. To understand how sampling effort affects the accuracy and precision of estimates, we applied each technique to six different-sized subsets of data per male. When averaged across subset sizes, the capture–recapture and coupon collector techniques showed the highest accuracy, whereas the curve fitting technique underestimated repertoire size. Precision (the average absolute difference between the estimated and true repertoire size) was significantly better for the capture–recapture technique than the coupon collector and curve fitting techniques. Both accuracy and precision improved as subset size increased. We conclude that capture–recapture is the best technique for estimating the sizes of small repertoires.     

Energetic reserves, leptin and testosterone: a refinement of the immunocompetence handicap hypothesis

The immunocompetence handicap hypothesis (ICHH) assumes that testosterone (T), required for the expression of sexual traits, can also incur a cost due to its immunosuppressive properties. However, T-dependent immunosuppression could also arise as an indirect consequence of energy reallocation from the immune system to other metabolic demands. Leptin is mostly produced in lipogenic tissues and its circulating level is positively correlated with the amount of lipid reserves. Leptin also has an important role as immunoenhancer and we suggest that this hormone could play a role as a mediator of the immunosuppressive effect of testosterone. In particular, we propose that only the individuals able to maintain large lipid reserves (with high leptin levels), while sustaining high testosterone levels, might be able to develop sexual displays without an impairment of their immune defences. Here, we tested one of the assumptions underlying this extension of the ICHH: leptin administration should attenuate testosterone-induced immunosuppression. T-implanted and control male zebra finches (Taeniopygia guttata) received daily injections of leptin or phosphate buffered saline. T-implants initially depressed the phytohaemagglutinin-induced immune response. However, T-birds injected with leptin enhanced their immune response to the level of control birds. These results open a new perspective on the study of the ICHH.     

Nestling testosterone is associated with begging behaviour and fledging success in the pied flycatcher, Ficedula hypoleuca

Animal signals are hypothesized to be costly in order to honestly reflect individual quality. Offspring solicitation signals given by nestling birds are thought to have evolved to advertise either need or individual quality. We tested the potential role of testosterone (T) in controlling the intensity of these signals by measuring begging behaviour as: (i) duration of the begging display and (ii) maximum height of the begging stretch, and by sampling endogenous T levels in nestling blood. We tested nestling pied flycatchers (Ficedula hypoleuca) using well-established experimental paradigm involving transient food deprivation to encourage begging behaviour and then blood-sampled nestlings at the end of these tests for T levels. Our results show that individual nestlings with the most intense begging displays had the highest circulating levels of T immediately after testing. In addition, we found substantial differences between broods in terms of circulating T. Finally, we found evidence that broods with higher levels of T showed increased fledging success, indicating a benefit for increased T production in nestlings. The potential trade-offs involved in T-mediated begging behaviour are discussed.     

Heritability of gonad size varies across season in a wild songbird

Many organisms advance their seasonal reproduction in response to global warming. In birds, which regress their gonads to a nonfunctional state each winter, these shifts are ultimately constrained by the time required for gonadal development in spring. Gonadal development is photoperiodically controlled and shows limited phenotypic plasticity in relation to environmental factors, such as temperature. Heritable variation in the time required for full gonadal maturation to be completed, based on both onset and speed of development and resulting in seasonally different gonad sizes among individuals, is thus a crucial prerequisite for an adaptive advancement of seasonal reproduction in response to changing temperatures. We measured seasonal gonadal development in climate‐controlled aviaries for 144 great tit (Parus major) pairs, which consisted of siblings obtained as whole broods from the wild. We show that the extent of ovarian follicle development (follicle size) in early spring is highly heritable (h² = 0.73) in females, but found no heritability of the extent of testis development in males. However, heritability in females decreased as spring advanced, caused by an increase in environmental variance and a decrease in additive genetic variation. This low heritability of the variation in a physiological mechanism underlying reproductive timing at the time of selection may hamper genetic adaptation to climate change, a key insight as this great tit population is currently under directional selection for advanced egg‐laying.     

Nestling testosterone controls begging behaviour in the pied flycatcher, Ficedula hypoleuca

Begging signals and endogenous testosterone (T) levels of young birds have been shown to be positively correlated. If T is causally involved in controlling the level of begging effort, an endocrine control mechanism could explain the evolution of begging as a costly signal reflecting need. We tested experimentally whether elevated circulating T levels enhanced begging behaviour in nestling pied flycatchers, Ficedula hypoleuca. A pilot study confirmed that nestling T levels could be elevated within a natural physiological range using an oral dose of T. After T-dosing, nestling begging behaviour was measured as: i) the duration of begging displays and ii) the maximum height of begging stretches. Our results show that nestling T levels were elevated at 90 min post dosing and that at this time point both measures of begging behaviour were performed more intensely by T-dosed nestlings than controls. Nestling begging displays in response to dosing varied between individuals, which in part was explained either by the date in the breeding season or nestling mass. The results of this study confirm the causal nature of T in controlling nestling begging signals and suggest that it may be part of the mechanism that controls begging behaviour in nestling birds.     

Population density, body size, and phenotypic plasticity of brood size in a burying beetle

Theory predicts that organisms living in heterogeneous environmentswill exhibit phenotypic plasticity. One trait that may be particularlyimportant in this context is the clutch or brood size becauseit is simultaneously a maternal and offspring characteristic.In this paper, I test the hypothesis that the burying beetle,Nicrophorus orbicollis, adjusts brood size, in part, in anticipationof the reproductive environment of its adult offspring. N. orbicollisuse a small vertebrate carcass as a food resource for theiryoung. Both parents provide parental care and actively regulatebrood size through filial cannibalism. The result is a positivecorrelation between brood size and carcass size. Adult bodysize is an important determinant of reproductive success forboth sexes, but only at higher population densities. I testthree predictions generated by the hypothesis that beetles adjustbrood size in response to population density. First, averageadult body size should vary positively with population density.Second, brood size on a given-sized carcass should be larger(producing more but smaller young) in low-density populationsthan in high-density populations. Third, females should respondadaptively to changes in local population density by producinglarger broods when population density is low and small broodswhen population density is high. All three predictions weresupported using a combination of field and laboratory experiments.These results (1) show that brood size is a phenotypically plastictrait and (2) support the idea that brood size decisions arean intergenerational phenomenon that varies with the anticipatedcompetitive environment of the offspring.     

Clutch size, brood size and brood emergence in the Marbled Teal Marmaronetta angustirostris in the Marismas del Guadalquivir, southwestern Spain

The first detailed study of the breeding biology of the globally threatened Marbled Teal Marmaronetta angustirostris is presented, using data from the Marismas del Guadalquivir in southwestern Spain. Complete clutches, with 9–20 eggs, were recorded from 28 April to 27 July. Excluding clutches of over 14 eggs (likely to be intraspecifically parasitized), the mean clutch size was 11.8, larger than that reported for almost all other Anatidae species. Broods hatched from May to early August (later than sympatric duck species), with a peak in mid-late June and significant annual variation in date. Brood sizes were exceptionally high, declining from a mean of 10.9 in the first week of age to 8.5 when fully feathered (at 42 days or more). Duckling mortality was concentrated in the first 2 weeks after hatching. Nine percent of broods had 15–23 ducklings, largely as a result of nest parasitism. Brood size decreased strongly with hatching date and varied significantly between years. Data from across the species' range showed that breeding occurs later at more northerly latitudes. A recent decline in the Marbled Teal in the Marismas is linked with its late breeding season. Anthropogenic changes have reduced the duration of flooding in the natural marshes of Donana National Park, which now dry out by August, before this species has completed breeding