Functional neurogenomic responses to acoustic threats, including a heterospecific referential alarm call and its referent, in the auditory forebrain of red-winged blackbirds.

In animal communication, functionally referential alarm calls elicit the same behavioral responses as their referents, despite their typically distinct bioacoustic traits. Yet the auditory forebrain in at least one songbird species, the black-capped chickadee Poecile atricapillus, responds similarly to threat calls and their referent predatory owl calls, as assessed by immediate early gene responses in the secondary auditory forebrain nuclei. Whether and where in the brain such perceptual and cognitive equivalence is processed remains to be understood in most other avian systems. Here, we studied the functional neurogenomic (non-) equivalence of acoustic threat stimuli perception by the red-winged blackbird Agelaius phoeniceus in response to the actual calls of the obligate brood parasitic brown-headed cowbird Molothrus ater and the referential anti-parasitic alarm calls of the yellow warbler Setophaga petechia, upon which the blackbird is known to eavesdrop. Using RNA-sequencing from neural tissue in the auditory lobule (primary and secondary auditory nuclei combined), in contrast to previous findings, we found significant differences in the gene expression profiles of both an immediate early gene, ZENK (egr-1), and other song-system relevant gene-products in blackbirds responding to cowbird vs. warbler calls. In turn, direct cues of threats (including conspecific intruder calls and nest-predator calls) elicited higher ZENK and other differential gene expression patterns compared to harmless heterospecific calls. These patterns are consistent with a perceptual non-equivalence in the auditory forebrain of adult male red-winged blackbirds in response to referential calls and the calls of their referents.

Host parent responses to heterospecific parasite nestling alarm calls are independent of past and current experience with experimental brood parasitism.

Communication between parents and dependent offspring is critical not only during provisioning, but also in antipredator contexts. In altricial birds, a top cause of reproductive failure is nest predation, and alarm calls both by parents and chicks can serve to alert others and increase the likelihood of offspring escaping predation. Understanding the factors that determine the strength of parental antipredator responses to different nestling alarm calls can provide insight into parent-offspring recognition. The prothonotary warbler (Protonotaria citrea), a host of the obligate brood parasite, the brown-headed cowbird (Molothrus ater), never rejects cowbird young and raises the parasite together with its own offspring. To determine whether warbler parents learn cowbird nestling alarm calls, we presented experimentally parasitized or non-parasitized parents with playbacks of conspecific warbler, parasitic cowbird, and a harmless heterospecific control, eastern bluebird (Sialis sialis), nestling alarm calls. We recorded the latency to respond and the number of chips given by members of the resident warbler pair. We found that parents were most likely to respond to warbler nestling alarm calls, least likely to respond to bluebird calls, with a statistically intermediate likelihood of responding to cowbird calls. Critically, current and past parasitism status did not affect the likelihood of response to any playback or the number of chips given, however, currently parasitized parents had greater response latencies to playbacks than non-parasitized parents. These results suggest that warbler parents do not learn cowbird alarm calls from breeding experiences and, in turn, that cowbirds may employ a generalized, bet-hedging alarm call.

The Direction of response selectivity between conspecific and heterospecific auditory stimuli varies with response metric.

Species recognition is an essential behavioral outcome of social discrimination, flocking, mobbing, mating, and/or parental care. In songbirds, auditory species recognition cues are processed through specialized forebrain circuits dedicated to acoustic discrimination. Here we addressed the direction of behavioral and neural metrics of zebra finches' (Taeniopygia guttata) responses to acoustic cues of unfamiliar conspecifics vs. heterospecifics. Behaviorally, vocal response rates were greater for conspecific male zebra finch songs over heterospecific Pin-tailed Whydah (Vidua macroura) songs, which paralleled greater multiunit spike rates in the auditory forebrain in response to the same type of conspecific over heterospecific auditory stimuli. In contrast, forebrain activation levels were reversed to species-specific song playbacks during two functional magnetic resonance imaging experiments: we detected consistently greater responses to whydah songs over finch songs and did so independently of whether subjects had been co-housed or not with heterospecifics. These results imply that the directionality of behavioral and neural response selectivity metrics are not always consistent and appear to be experience-independent in this set of stimulus-and-subject experimental paradigms.

Early acoustic experience alters genome-wide methylation in the auditory forebrain of songbird embryos.

Early exposure to salient cues can critically shape the development of social behaviors. For example, both oscine birds and humans can hear and learn to recognize familiar sounds in ovo and in utero and recognize them following hatching and birth, respectively. Here we demonstrate that different chronic acoustic playbacks alter genome-wide methylation of the auditory forebrain in late-stage zebra finch (Taeniopygia guttata) embryos. Within the same subjects, immediate early gene activation in response to acute con- or heterospecific song exposure is negatively correlated with methylation extent in response to repeated daily prior exposure to the same type of stimuli. Specifically, we report less relative global methylation following playbacks of conspecific songs and more methylation following playbacks of distantly-related heterospecific songs. These findings offer a neuroepigenomic mechanism for the ontogenetic impacts of early acoustic experiences in songbirds.

Exposure to a mimetic or non-mimetic model avian brood parasite egg does not produce differential glucocorticoid responses in an egg-accepter host species.

Avian obligate brood parasitism, a reproductive strategy where a parasite lays its egg into the nest of another species, imposes significant fitness costs upon host parents and their offspring. To combat brood parasitism, many host species recognize and reject foreign eggs (rejecters), but others are accepters that raise the parasitic progeny. Some accepter hosts may be unable to grasp or pierce parasitic eggs even if they recognize them as foreign eggs in the clutch, whereas other accepters may not have evolved the cognitive skillsets to recognize dissimilar eggs in the nest. Here we assessed the endocrine responses of an accepter host species to model parasitic eggs to address these two alternatives. We experimentally parasitized nests of a locally common host of the brood-parasitic brown-headed cowbird (Molothrus ater), the prothonotary warbler (Protonotaria citrea; a cowbird-egg accepter), with a mimetic or non-mimetic model cowbird-sized egg. Our goal was to determine whether they perceived the non-mimetic egg as a greater stressor by measuring circulating corticosterone levels. We added eggs to nests during the incubation stage and obtained blood plasma samples from females on the nest 2 h later, using females with unmanipulated clutches as controls. Incubating females showed no differences in baseline plasma corticosterone levels between our different treatments. We conclude that exposure to foreign eggs does not activate the hypothalamic-pituitary-adrenal axis of prothonotary warbler hosts in this experimental paradigm.

Inter-Individual Variation in Anti-Parasitic Egg Rejection Behavior: A Test of the Maternal Investment Hypothesis.

Hosts of avian brood parasites may reduce or forego the costs of caring for foreign young by rejecting parasitic eggs from the nest. Yet, many host species accept parasitic eggs and, even among rejecter species, some individuals go on to incubate and hatch them. The factors explaining the variation in egg rejection between species have received much theoretical and empirical attention, but the causes of intraspecific variation in different individuals' propensity for accepting parasitic eggs are less well understood. Here we tested the maternal investment hypothesis, which predicts that hosts with costlier clutches will be more likely to reject parasitic eggs from their nest. We studied variation in the egg rejection responses of American robins (Turdus migratorius), a robust egg-rejecter host of the brood parasitic brown-headed cowbird (Molothrus ater), to 3D-printed cowbird-sized eggs which were painted dark blue, a color known to induce variable and repeatable egg rejection responses in individual robins. Costlier clutch investment was estimated by earlier laying date, larger clutch size, heavier unincubated yolk mass, and variable yolk steroid hormone concentrations. There was no statistical support for most of our predictions. However, we detected more concentrated and greater overall amount of deoxycorticosterone deposited in egg yolks of rejecters relative to acceptors, although this accounted for no more than 14% of variance in the data. Future work should test experimentally the potential physiological linkage between maternal egg yolk steroid investment and egg rejection propensity in this and other host species of avian brood parasites.

Egyedek közti variáció az antiparazitikus tojás-diszkriminációban: az anyai befektetés hipotézis tesztelése Kivonat A költésparazita madárfajok gazdái csökkenthetik vagy megszüntethetik az idegen fióka nevelésének költségeit úgy, hogy eltávolítják a fészekbol a paraziták tojásait. Ennek ellenére több gazdafaj is elfogadja a parazita tojásokat, sot az elutasító gazdafajok körében is akadnak olyan egyedek, amelyek elfogadják és kikeltik az idegen tojást. Bár az eddigi elméleti és kísérleti kutatások nagy hangsúlyt fektettek a különbözo gazdafajok közti, költésparazita tojások diszkriminacióját befolyásoló tényezok feltárására, a parazita tojások elfogadásának gazdafajon belüli varianciája kevésbé tisztázott. Jelen kutatásunkban az anyai befektetés hipotézisét vizsgáltuk, melynek értelmében azok az egyedek, amelyek több energiát fektetnek fészekaljukba, nagyobb valószínuséggel távolítják el a költésparazita tojásait a fészekbol. A vándorrigónak (Turdus migratorius) a parazita tojást elutasító viselkedését vizsgáltuk meg a költésparazita barnafeju gulyajáró (Molothrus ater) tojásához hasonló méretu, de sötétkék színu, 3 D nyomtatóval készített mutojásokat használva. A költési periódusban korábban letojt, több tojásból álló, nagyobb inkubálatlan tömegu és változatos szteroid hormonkoncentrácijú sárga szíku tojásokkal rendelkezo fészekaljakat tekintettük a magasabb anyai befektetésu fészekaljaknak. Feltevéseink nagyrészét statisztikai eredményeink nem támasztották alá. Mindezek ellenére, a parazita tojásokat elfogadó egyedekéhez képest, az idegen tojást elutasító egyedek tojásai sárga szíkanyagában nagyobb koncentrációban és mennyiségben volt jelen a dezoxikortikoszteron, bár ez az adatainkra jellemzo varianciának csupán 14%-át magyarázta. További kísérletes vizsgálatok szükségesek ahhoz, hogy feltárhassuk az esetleges fiziológiás kapcsolatot az anyai tojas szteroid hormone befektetés és a parazita tojások elutasításának gyakorisága között ennél a gazdafajnál, és más költésparazita gazdáknál egyaránt. Translated kindly by Attila Marton, Debrecen University.

Parasitic egg rejection decisions of chalk-browed mockingbirds Mimus saturninus are independent of clutch composition.

Obligate avian brood parasites lay their eggs in nests of other host species, which assume all the costs of parental care for the foreign eggs and chicks. The most common defensive response to parasitism is the rejection of foreign eggs by hosts. Different cognitive mechanisms and decision-making rules may guide both egg recognition and rejection behaviors. Classical optimization models generally assume that decisions are based on the absolute properties of the options (i.e., absolute valuation). Increasing evidence shows instead that hosts' rejection decisions also depend on the context in which options are presented (i.e., context-dependent valuation). Here we study whether the chalk-browed mockingbird's (Mimus saturninus) rejection of parasitic shiny cowbird (Molothrus bonariensis) eggs is a fixed behavior or varies with the context of the clutch. We tested three possible context-dependent mechanisms: (1) range effect, (2) habituation to variation, and (3) sensitization to variation. We found that mockingbird rejection of parasitic eggs does not change according to the characteristics of the other eggs in the nest. Thus, rejection decisions may exclusively depend on the objective characteristics of the eggs, meaning that the threshold of acceptance or rejection of a foreign egg is context-independent in this system.

Analyses of sex and individual differences in vocalizations of Australasian gannets using a dynamic time warping algorithm.

The study of the evolution of sexual differences in behavioral and morphological displays requires analyses of the extent of sexual dimorphism across various sensory modalities. In the seabird family Sulidae, boobies show dramatic sexual dimorphism in their vocalizations, and gannet calls have also been suggested to be dimorphic to human observers. This study aimed to evaluate the presence of sexually dimorphic calls in the Australasian gannet (Morus serrator) through the first comprehensive description of its vocalizations recorded at two localities; Cape Kidnappers, where individuals were banded and sexed from DNA samples, and at the Muriwai gannetry, both on the North Island of New Zealand. Calls were first inspected using basic bioacoustic features to establish a library of call element types for general reference. Extensive multivariate tests, based on a dynamic time warping algorithm, subsequently revealed that no sexual differences could be detected in Australasian gannet calls. The analyses, however, indicated extensive and consistent vocal variation between individuals, particularly so in female gannets, which may serve to signal individual identity to conspecifics. This study generates predictions to identify whether differences in Australasian gannet vocalizations play perceptual and functional roles in the breeding and social biology of this long-lived biparental seabird species.

Experimental support for the role of nest predation in the evolution of brood parasitism.

In 1965, Hamilton and Orians (HO) hypothesized that the starting point for the evolution of obligate interspecific brood parasitism in birds was the facultative laying of physiologically committed eggs in neighbouring active nests of con- and heterospecifics, following predation of a bird's own nest during the laying stage. We tested this prediction of the HO hypothesis by using captive pairs of zebra finches (Taeniopygia guttata), a species with evidence for intraspecific parasitism both in the wild and in captivity. As predicted, in response to experimental nest removal, subjects laid eggs parasitically in simulated active conspecific nests above chance levels. Across subsequent trials, we detected both repeatability and directional change in laying patterns, with some subjects switching from parasitism to depositing eggs in the empty nest. Taken together, these results support the assumptions and predictions of the HO hypothesis, and indicate that the zebra finch is a potential model species for future behavioural and genetic studies in captive brood parasite research.

To eject or to abandon? Life history traits of hosts and parasites interact to influence the fitness payoffs of alternative anti-parasite strategies.

Hosts either tolerate avian brood parasitism or reject it by ejecting parasitic eggs, as seen in most rejecter hosts of common cuckoos, Cuculus canorus, or by abandoning parasitized clutches, as seen in most rejecter hosts of brown-headed cowbirds, Molothrus ater. What explains consistent variation between alternative rejection behaviours of hosts within the same species and across species when exposed to different types of parasites? Life history theory predicts that when parasites decrease the fitness of host offspring, but not the future reproductive success of host adults, optimal clutch size should decrease. Consistent with this prediction, evolutionarily old cowbird hosts, but not cuckoo hosts, have lower clutch sizes than related rarely- or newly parasitized species. We constructed a mathematical model to calculate the fitness payoffs of egg ejector vs. nest abandoner hosts to determine if various aspects of host life history traits and brood parasites' virulence on adult and young host fitness differentially influence the payoffs of alternative host defences. These calculations showed that in general egg ejection was a superior anti-parasite strategy to nest abandonment. Yet, increasing parasitism rates and increasing fitness values of hosts' eggs in both currently parasitized and future replacement nests led to switch points in fitness payoffs in favour of nest abandonment. Nonetheless, nest abandonment became selectively more favourable only at lower clutch sizes and only when hosts faced parasitism by a cowbird- rather than a cuckoo-type brood parasite. We suggest that, in addition to evolutionary lag and gape-size limitation, our estimated fitness differences based on life history trait variation provide new insights for the consistent differences observed in the anti-parasite rejection strategies between many cuckoo- and cowbird-hosts.

Self-referent phenotype matching: theoretical considerations and empirical evidence.

In most birds and mammals, young are raised in family groups. The phenotypes of nestmates and parents are thus reliable cues for recognition of conspecifics and kin. However, in some species, young develop alone, or in broods of mixed relatedness (e.g. because of multiple paternity or maternity), or among heterospecifics or unrelated conspecifics (brood parasites). Under these circumstances, the best referent (model) for discriminating close from distant kin and heterospecifics from conspecifics might be one's own self. This recognition process is known as self-referent phenotype matching. Here we review recent experimental evidence of self-referencing and suggest that behavioral neuroscience can provide new tools and insights into how it works (its proximate mechanistic and ontogenetic bases) and why it exists (its adaptive significance).

Signal polymorphism in the web-decorating spider Argiope argentata is correlated with reduced survivorship and the presence of stingless bees, its primary prey.

Many spiders, and in particular those in the genus Argiope, spin highly visible web decorations whose function and significance are the subject of spirited debate. In this work, we present data to address two of the competing hypotheses that fuel this controversy. In particular, we examine the relationship between the presence of web decorations and spider survivorship (predator-protection hypothesis) and the relationship between the presence of prey and spider decorating behavior (the prey-attraction hypothesis). Our laboratory studies reveal that the decorating behavior of the spider A. argentata has a genetic component but that the expression of decorating behavior tends to be elicited only when a spider is well fed. Furthermore, our field studies show that in the presence of abundant stingless bees, spider decorating behavior is induced. Nevertheless, our field surveys also suggest that spiders that decorate their webs show reduced survivorship. We propose that the high correlation between web decorating in the presence of stingless bees supports the hypothesis that A. argentata engage in decorating behavior when attracting or targeting specific prey types. However, we also propose that web decorations attract the predators of A. argentata because high-frequency decorators suffer lower survivorship than spiders that decorate moderately or rarely. These findings suggest that spider web decorating behavior is affected by conflicting selection pressures: the positive effect of prey attraction versus the negative effect of predator attraction. Due to the heritable component of decorating behavior, web decorating among A. argentata is likely to be particularly sensitive to the spider's local ecology as well as local patterns of gene flow.

A password for species recognition in a brood-parasitic bird.

Recognition of conspecifics is an essential precursor of sexual reproduction. Most mammals and birds learn salient features of their parents or siblings early in ontogeny and later recognize individuals whose phenotypes match the mental image (template) of relatives closely enough as conspecifics. However, the young of brood parasites are reared among heterospecifics, so social learning will yield inappropriate species recognition templates. Initially, it was inferred that conspecific recognition in brood parasites depended on genetically determined templates. More recently it was demonstrated that learning plays a critical role in the development of parasites' social preferences. Here we propose a mechanism that accommodates the interaction of learned and genetic components of recognition. We suggest that conspecific recognition is initiated when a young parasite encounters some unique species-specific signal or "password" (e.g. a vocalization, behaviour or other characteristic) that triggers learning of additional aspects of the password-giver's phenotype. We examined the possibility that nestlings of the obligately brood-parasitic brown-headed cowbird (Molothrus ater) could use a species-specific vocalization, the "chatter", as a password. We found that six-day-old nestlings responded (begged) significantly more frequently to playbacks of chatters than to other avian sounds and that two-month-old fledglings approached playbacks of chatters more quickly than vocalizations of heterospecifics. Free-living cowbird fledglings and adults also approached playbacks of chatters more often than control sounds. Passwords may be involved in the ontogeny of species recognition in brood parasites generally.

Sexual dimorphism and species differences in HVC volumes of cowbirds.

Cowbirds exhibit extensive variation in their social, territorial, and reproductive behaviors. Nissl-stained brain sections of specimens from a previous study (J. C. Reboreda, N. S. Clayton, & A. Kacelnik, 1996) were used to study the gross anatomy of a song control nucleus in 3 South American cowbirds (bay-winged, Molothrus badius; shiny, M. bonariensis; and screaming, M. rufoaxillaris). Cowbird high vocal center (HVC) volumes were consistently higher in males than in females in all 3 species. The largest HVC size of females found in bay-winged cowbirds is consistent with observations that females of this species, but not of the other 2 species, occasionally sing. The extent of the sexual dimorphism of relative HVC size was highest for the sexually dichromatic and promiscuous shiny cowbirds and smaller for the monochromatic and monogamous bay-winged and screaming cowbirds, suggesting that selection pressures associated with morphological traits and social systems are reflected in brain architecture.

The effect of radar on cardiac pacemakers.

The susceptibility of 16 noncompetitive cardiac pacemakers to radiation from a powerful radar system was investigated in the laboratory and in the vicinity of its prototype. From comparative in vitro tests in air, fat, water, and saline it was concluded that only tests in fat or air represent the worst case condition after implantation. In air all pacemakers showed signs of interference at pulse power densities between 0.025 mW/cm2 and 62.5 mW/CM2. Three of six implanted pacemakers were triggered or inhibited depending on their mode of operation when tested at a location 1.2 km away from the radar station by the radar beam occurring every 5.5 sec. Because interfering radiation can enter the pacemaker circuitry directly along the electrode, acting as an antenna, metal encapsulation of the pulse generator does not provide sufficient shielding against microwave radiation. However, pacemakers modified by metal encapsulation and a low-pass filter at the electrode remained undistrubed at pulse power densities of greater than 10 W/cm2 when tested under worst case condition in air.