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Vocal rhythm plays a fundamental role in sexual selection and species recognition in birds, but little is known of its genetic basis due to the confounding effect of vocal learning in model systems. Uncovering its genetic basis could facilitate identifying genes potentially important in speciation. Here we investigate the genomic underpinnings of rhythm in vocal non-learning Pogoniulus tinkerbirds using 135 individual whole genomes distributed across a southern African hybrid zone. We find rhythm speed is associated with two genes that are also known to affect human speech, Neurexin-1 and Coenzyme Q8A. Models leveraging ancestry reveal these candidate loci also impact rhythmic stability, a trait linked with motor performance which is an indicator of quality. Character displacement in rhythmic stability suggests possible reinforcement against hybridization, supported by evidence of asymmetric assortative mating in the species producing faster, more stable rhythms. Because rhythm is omnipresent in animal communication, candidate genes identified here may shape vocal rhythm across birds and other vertebrates., (© 2024. The Author(s).)

Birds remodel their flight muscle metabolism prior to migration to meet the physiological demands of migratory flight, including increases in both oxidative capacity and defence against reactive oxygen species. The degree of plasticity mediated by changes in these mitochondrial properties is poorly understood but may be explained by two non-mutually exclusive hypotheses: variation in mitochondrial quantity or in individual mitochondrial function. We tested these hypotheses using yellow-rumped warblers (Setophaga coronata), a Nearctic songbird which biannually migrates 2000-5000 km. We predicted higher flight muscle mitochondrial abundance and substrate oxidative capacity, and decreased reactive oxygen species emission in migratory warblers captured during autumn migration compared with a short-day photoperiod-induced non-migratory phenotype. We assessed mitochondrial abundance via citrate synthase activity and assessed isolated mitochondrial function using high-resolution fluororespirometry. We found 60% higher tissue citrate synthase activity in the migratory phenotype, indicating higher mitochondrial abundance. We also found 70% higher State 3 respiration (expressed per unit citrate synthase) in mitochondria from migratory warblers when oxidizing palmitoylcarnitine, but similar H2O2 emission rates between phenotypes. By contrast, non-phosphorylating respiration was higher and H2O2 emission rates were lower in the migratory phenotype. However, flux through electron transport system complexes I-IV, II-IV and IV was similar between phenotypes. In support of our hypotheses, these data suggest that flight muscle mitochondrial abundance and function are seasonally remodelled in migratory songbirds to increase tissue oxidative capacity without increasing reactive oxygen species formation., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Trees release Herbivore-Induced Plant Volatiles (HIPVs) into the air in response to damage inflicted by insects. It is known that songbirds use those compounds to locate their prey, but more recently the idea emerged that songbirds could also use those odours as cues in their reproductive decisions, as early spring HIPVs may contain information about the seasonal timing and abundance of insects. We exposed pre-breeding great tits (Parus major) to the odours of caterpillar-infested trees under controlled conditions, and monitored reproduction (timing of egg laying, number of eggs, egg size) and two of its main hormonal drivers (testosterone and 17β-estradiol in males and females, respectively). We found that females exposed to HIPVs did not advance their laying dates, nor laid larger clutches, or larger eggs compared to control females. 17β-estradiol concentrations in females were also similar between experimental and control birds. However, males exposed to HIPVs had higher testosterone concentrations during the egg-laying period. Our study supports the hypothesis that insectivorous songbirds are able to detect minute amounts of plant odours. The sole manipulation of plant scents was not sufficient to lure females into a higher reproductive investment, but males increased their reproductive effort in response to a novel source of information for seasonal breeding birds., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2024. Published by Elsevier Inc.)

The auditory sensitivity of a small songbird, the red-cheeked cordon bleu, was measured using the standard methods of animal psychophysics. Hearing in cordon bleus is similar to other small passerines with best hearing in the frequency region from 2 to 4 kHz and sensitivity declining at the rate of about 10 dB/octave below 2 kHz and about 35 dB/octave as frequency increases from 4 to 9 kHz. While critical ratios are similar to other songbirds, the long-term average power spectrum of cordon bleu song falls above the frequency of best hearing in this species., (© 2024 Acoustical Society of America.)

A significant body of evidence indicates that climate change is influencing many aspects of avian ecology. Yet, how climate change is affecting, and is expected to influence some aspects of the breeding ecology of cavity-nesting birds remains uncertain. To explore the potential linkage between timing of first clutch, and the influence of ambient temperature on hatching success, we used Eastern Bluebird ( Sialia sialis ) nest records over a nine-year period from Alabama, USA. We investigated changes to annual clutch initiation dates, as well as variability in hatching success associated with ambient air temperatures during the incubation period. Using a simple linear model, we observed earlier annual egg laying dates over the nine years of this study with a difference of 24 days between earliest egg-laying date of the season. Daily temperature minima increased 2 °C across the nine-year time frame of this study. These data also indicate that Eastern Bluebird hatching success was the highest when mean ambient air temperature during incubation was between 19 °C and 24 °C (78%, as opposed to 69% and 68% above and below this temperature range, respectively). Our findings of increasing maxima, earlier maxima each year, and the lower minima of temperatures within our study area could expand the breadth of temperatures experienced by nesting Eastern Bluebirds possibly exposing them to temperatures outside of what promotes nesting success. These findings with a cavity-nesting bird highlight an optimal range of ambient temperatures associated with highest hatching success, conditions likely to be affected by climate change., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Throughout the life of the adult songbird, neurons are recruited into brain regions important for song learning. Movies captured by Shvedov et al. demonstrate this dynamic process in the live animal, revealing the mechanisms of neuronal migration in the adult brain., Competing Interests: Declaration of interests The author declares no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Aggression is a social behavior that is critical for survival and reproduction. In adults, circulating gonadal hormones, such as androgens, act on neural circuits to modulate aggressive interactions, especially in reproductive contexts. In many species, individuals also demonstrate aggression before reaching gonadal maturation. Adult male song sparrows, Melospiza melodia, breed seasonally but maintain territories year-round. Juvenile (hatch-year) males aggressively compete for territory ownership during their first winter when circulating testosterone is low. Here, we characterized the relationship between the steroid milieu and aggressive behavior in free-living juvenile male song sparrows in winter. We investigated the effect of a 10 min simulated territorial intrusion (STI) on behavior and steroid levels in blood, 10 microdissected brain regions, and four peripheral tissues (liver, pectoral muscle, adrenal glands, and testes). Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we quantified 12 steroids: pregnenolone, progesterone, corticosterone, 11-dehydrocorticosterone, dehydroepiandrosterone, androstenedione, testosterone, 5α-dihydrotestosterone, 17β-estradiol , 17α-estradiol, estrone, and estriol. We found that juvenile males are robustly aggressive, like adult males. An STI increases progesterone and corticosterone levels in blood and brain and increases 11-dehydrocorticosterone levels in blood only. Pregnenolone, androgens, and estrogens are generally non-detectable and are not affected by an STI. In peripheral tissues, steroid concentrations are very high in the adrenals. These data suggest that adrenal steroids, such as progesterone and corticosterone, might promote juvenile aggression and that juvenile and adult songbirds might rely on distinct neuroendocrine mechanisms to support similar aggressive behaviors., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Males usually come into conflict due to competition for territories and females. However, interference competition can also occur between males of congeneric species when their ecological requirements are overlapping. Using acoustic playback experiments, we investigated male-male interactions within and between Grasshopper (Locustella naevia; GW) and River Warbler (L. fluviatilis; RW). Our objective was to evaluate the song and behavioural response of tested males of both species to conspecific song stimuli in order to compare this with the response to congeneric stimulus, based on which we could assess whether these two commonly co-existing species show interspecific territorialism. A total of nine GW and 11 RW males were tested in May and June 2019 in western Slovakia. The ability to differentiate between the heterospecific (control), congeneric, and conspecific stimuli was similar between the two species. Conspecific playback elicited the strongest non-vocal response and a significant change in vocalization. The GW males shortened the songs, while the RW males shortened the songs and also increased their syllable rate. The congeneric playback elicited a lower intensity of behavioural response than conspecific playback and no change in vocalization in either species. We conclude that interspecific interference competition between GW and RW is rather low, suggesting that the species' ecological requirements are separated, although these two congeneric species commonly share habitat., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Migration is an energetically challenging and risky life history stage for many animals, but could be supported by dietary choices en route, which may create opportunities to improve body and physiological condition. However, proposed benefits of diet shifts, such as between seasonally available invertebrates and fruits, have received limited investigation in free-living animals. We quantified diet composition and magnitude of autumn diet shifts over two time periods in two closely-related species of migratory songbirds on stopover in the northeastern U.S. (Swainson's thrush [Catharus ustulatus], long-distance migrant, N = 83; hermit thrush [C. guttatus], short-distance migrant, N = 79) and used piecewise structural equation models to evaluate the relationships among (1) migration timing, (2) dietary behavior, and (3) morphometric and physiological condition indices. Tissue isotope composition indicated that both species shifted towards greater fruit consumption. Larger shifts in recent weeks corresponded to higher body condition in Swainson's, but not hermit thrushes, and condition was more heavily influenced by capture date in Swainson's thrushes. Presence of "high-antioxidant" fruits in fecal samples was unrelated to condition in Swainson's thrushes and negatively related to multiple condition indices in hermit thrushes, possibly indicating the value of fruits during migration is related more to their energy and/or macronutrient content than antioxidant content. Our results suggest that increased frugivory during autumn migration can support condition, but those benefits might depend on migration strategy: a longer-distance, more capital-dependent migration strategy could require stricter regulation of body condition aided by increased fruit consumption., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Neuron migration is a key phase of neurogenesis, critical for the assembly and function of neuronal circuits. In songbirds, this process continues throughout life, but how these newborn neurons disperse through the adult brain is unclear. We address this question using in vivo two-photon imaging in transgenic zebra finches that express GFP in young neurons and other cell types. In juvenile and adult birds, migratory cells are present at a high density, travel in all directions, and make frequent course changes. Notably, these dynamic migration patterns are well fit by a superdiffusive model. Simulations reveal that these superdiffusive dynamics are sufficient to disperse new neurons throughout the song nucleus HVC. These results suggest that superdiffusive migration may underlie the formation and maintenance of nuclear brain structures in the postnatal brain and indicate that transgenic songbirds are a useful resource for future studies into the mechanisms of adult neurogenesis., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

We investigated within- and between-individual song variation and song-based neighbour-stranger discrimination in a non-learning bird species, the blue-headed wood-dove (Turtur brehmeri), which inhabits lowland rainforests of West and Central Africa. We found that songs of this species are individually specific and have a high potential for use in individual recognition based on the time-frequency pattern of note distribution within song phrases. To test whether these differences affect behaviour, we conducted playback experiments with 19 territorial males. Each male was tested twice, once with the songs of a familiar neighbour and once with the songs of an unfamiliar stranger. We observed that males responded more aggressively to playback of a stranger's songs: they quickly approached close to the speaker and spent more time near it. However, no significant differences between treatments were observed in the vocal responses. In addition, we explored whether responses differed based on the song frequency of the focal male and/or that of the simulated intruder (i.e., playback), as this song parameter is inversely related to body size and could potentially affect males' decisions to respond to other birds. Song frequency parameters (of either the focal male or the simulated intruder) had no effect on the approaching response during playback. However, we found that the pattern of response after playback was significantly affected by the song frequency of the focal male: males with lower-frequency songs stayed closer to the simulated intruder for a longer period of time without singing, while males with higher-frequency songs returned more quickly to their initial song posts and resumed singing. Together, these results depict a consistently strong response to strangers during and after playback that is dependent on a male's self-assessment rather than assessment of a rival's strength based on his song frequency. This work provides the first experimental evidence that doves (Columbidae) can use songs for neighbour-stranger discrimination and respond according to a "dear enemy" scheme that keeps the cost of territory defence at a reasonable level., (© 2024. The Author(s).)

Migratory birds possess remarkable accuracy in orientation and navigation, which involves various compass systems including the magnetic compass. Identifying the primary magnetosensor remains a fundamental open question. Cryptochromes (Cry) have been shown to be magnetically sensitive, and Cry4a from a migratory songbird seems to show enhanced magnetic sensitivity in vitro compared to Cry4a from resident species. We investigate Cry and their potential involvement in magnetoreception in a phylogenetic framework, integrating molecular evolutionary analyses with protein dynamics modelling. Our analysis is based on 363 bird genomes and identifies different selection regimes in passerines. We show that Cry4a is characterized by strong positive selection and high variability, typical characteristics of sensor proteins. We identify key sites that are likely to have facilitated the evolution of an optimized sensory protein for night-time orientation in songbirds. Additionally, we show that Cry4 was lost in hummingbirds, parrots and Tyranni (Suboscines), and thus identified a gene deletion, which might facilitate testing the function of Cry4a in birds. In contrast, the other avian Cry (Cry1 and Cry2) were highly conserved across all species, indicating basal, non-sensory functions. Our results support a specialization or functional differentiation of Cry4 in songbirds which could be magnetosensation.

AbstractVocal production learning (the capacity to learn to produce vocalizations) is a multidimensional trait that involves different learning mechanisms during different temporal and socioecological contexts. Key outstanding questions are whether vocal production learning begins during the embryonic stage and whether mothers play an active role in this through pupil-directed vocalization behaviors. We examined variation in vocal copy similarity (an indicator of learning) in eight species from the songbird family Maluridae, using comparative and experimental approaches. We found that (1) incubating females from all species vocalized inside the nest and produced call types including a signature "B element" that was structurally similar to their nestlings' begging call; (2) in a prenatal playback experiment using superb fairy wrens ( Malurus cyaneus ), embryos showed a stronger heart rate response to playbacks of the B element than to another call element (A); and (3) mothers that produced slower calls had offspring with greater similarity between their begging call and the mother's B element vocalization. We conclude that malurid mothers display behaviors concordant with pupil-directed vocalizations and may actively influence their offspring's early life through sound learning shaped by maternal call tempo.

Achieving endothermic homeothermy is a critical aspect of avian development. In pre-homeothermic altricial nestlings, variation in parental brooding behavior results in variable exposure of nestlings to cooling, with consequences for the developing endocrine system. Nestlings facing repeated cooling challenges may benefit from upregulation of thyroid hormone secretion, allowing for earlier onset of thermoregulatory capability to mitigate the potentially negative effects of exposure to non-optimal temperatures during development. We examined the effects of (1) a single cooling challenge on thyroid hormone secretion in pre-homeothermic nestlings, and (2) repeated cooling challenges prior to the onset of homeothermy on nestling growth and thyroid hormone secretion prior to fledging. We found that pre-homeothermic eastern bluebird nestlings exposed to a single cooling challenge increased circulating triiodothyronine (T 3 ), demonstrating that the thyroid system can be activated by cooling early in life. However, we found no consequences of repeated cooling during the first week of life on nestling growth or baseline T 3 levels prior to fledging. This work addresses how the nestling hypothalamic-pituitary-thyroid axis responds to acute cooling challenges prior to the development of endothermic homeothermy; future work will confirm whether such responses allow nestlings to hasten the onset of physiological thermoregulation when conditions demand it., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

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., (© 2024. The Author(s).)

Many vertebrates eavesdrop on alarm calls of other species, as well as responding to their own species' calls, but eavesdropping on heterospecific alarm calls might be harder than conspecific reception when environmental conditions make perception or recognition of calls difficult. This could occur because individuals lack hearing specializations for heterospecific calls, have less familiarity with them, or require more details of call structure to identify calls they have learned to recognize. We used a field playback experiment to provide a direct test of whether noise, as an environmental perceptual challenge, reduces response to heterospecific compared to conspecific alarm calls. We broadcast superb fairy-wren ( Malurus cyaneus ) and white-browed scrubwren ( Sericornis frontalis ) flee alarm calls to each species with or without simultaneous broadcast of ambient noise. Using two species allows isolation of the challenge of heterospecific eavesdropping independently of any effect of call structure on acoustic masking. As predicted, birds were less likely to flee to heterospecific than conspecific alarm calls during noise. We conclude that eavesdropping was harder in noise, which means that noise could disrupt information on danger in natural eavesdropping webs and so compromise survival. This is particularly significant in a world with increasing anthropogenic noise.

Carolina chickadees (Poecile carolinensis) and tufted titmice (Baeolophus bicolor) regularly form flocks with multiple species through the winter months, including white-breasted nuthatches (Sitta carolinensis). Earlier studies found that behavior of both chickadees and titmice was sensitive to mixed-species flock composition. Little is known about the influence of background noise level and vegetation density on the antipredator behaviors of individuals within these flocks, however. We tested for the effects of vegetation density, traffic noise, and flock composition (conspecific number, flock diversity, and flock size) on antipredator behavioral responses following an alarm call playback (Study 1) and an owl model presentation (Study 2) at feeders. We recorded background traffic noise and performed lidar scans to quantify vegetation density at each site. After a feeder had been stocked with seed and a flock was present, we recorded calls produced, and we identified flock composition metrics. We coded seed-taking latency, call latency, mob latency, and mob duration following the respective stimulus presentation and tested for effects of flock composition metrics, vegetation density, and background noise on these responses. For the alarm call playback study, flock composition drove behaviors in chickadees and titmice, and vegetation density drove behaviors in chickadees and nuthatches. For the owl model study, conspecific number predicted behavior in chickadees, and mob duration was predicted by nuthatch number. The results reveal individual sensitivity to group composition in anti-predatory and foraging behavior in simulated risky contexts. Additionally, our data suggest that the modality of perceived simulated risk (acoustic vs. visual) and the density of vegetation influence behavior in these groups., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Adams et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Currently, it is generally assumed that migratory birds are oriented in the appropriate migratory direction under UV, blue and green light (short-wavelength) and are unable to use their magnetic compass in total darkness and under yellow and red light (long-wavelength). However, it has also been suggested that the magnetic compass has two sensitivity peaks: in the short and long wavelengths, but with different intensities. In this project, we aimed to study the orientation of long-distance migrants, pied flycatchers ( Ficedula hypoleuca ), under different narrowband light conditions during autumn and spring migrations. The birds were tested in the natural magnetic field (NMF) and a changed magnetic field (CMF) rotated counterclockwise by 120° under dim green (autumn) and yellow (spring and autumn) light, which are on the 'threshold' between the short-wavelength and long-wavelength light. We showed that pied flycatchers (i) were completely disoriented under green light both in the NMF and CMF but (ii) showed the migratory direction in the NMF and the appropriate response to CMF under yellow light. Our data contradict the results of previous experiments under narrowband green and yellow light and raise doubts about the existence of only short-wavelength magnetoreception. The parameters of natural light change dramatically in spectral composition and intensity after local sunset, and the avian magnetic compass should be adapted to function properly under such constantly changing light conditions.

Vocal signals, including human speech and birdsong, are produced by complicated, precisely coordinated body movements, whose execution is fitness-determining in resource competition and mate choice. While the acquisition and maintenance of motor skills generally requires practice to develop and maintain both motor circuitry and muscle performance, it is unknown whether vocal muscles, like limb muscles, exhibit exercise-induced plasticity. Here, we show that juvenile and adult zebra finches (Taeniopygia castanotis) require daily vocal exercise to first gain and subsequently maintain peak vocal muscle performance. Experimentally preventing male birds from singing alters both vocal muscle physiology and vocal performance within days. Furthermore, we find females prefer song of vocally exercised males in choice experiments. Vocal output thus contains information on recent exercise status, and acts as an honest indicator of past exercise investment in songbirds, and possibly in all vocalising vertebrates., (© 2023. The Author(s).)

Diverse dopamine (DA) pathways send distinct reinforcement signals to different striatal regions. In adult songbirds, a DA pathway from the ventral tegmental area (VTA) to Area X, the striatal nucleus of the song system, carries singing-related performance error signals important for learning. Meanwhile, a parallel DA pathway to a medial striatal area (MST) arises from a distinct group of neighboring DA neurons that lack connectivity to song circuits and do not encode song error. To test if the structural and functional segregation of these two pathways depends on singing experience, we carried out anatomical studies early in development before the onset of song learning. We find that distinct VTA neurons project to either Area X or MST in juvenile birds before the onset of substantial vocal practice. Quantitative comparisons of early juveniles (30-35 days post hatch), late juveniles (60-65 dph), and adult (>90 dph) brains revealed an outsized expansion of Area X-projecting neurons relative to MST-projecting neurons in VTA over development. These results show that a mesostriatal DA system dedicated to social communication can exist and be spatially segregated before the onset of vocal practice and associated sensorimotor experience., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ramarao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The avian magnetic compass can be disrupted by weak narrow-band and broadband radio-frequency (RF) fields in the lower MHz range. However, it is unclear whether disruption of the magnetic compass results from the elimination of the perception pattern produced by the magnetic field or from qualitative changes that make the pattern unrecognizable. We show that zebra finches trained in a 4-arm maze to orient relative to the magnetic field are disoriented when tested in the presence of low-level (~ 10 nT) Larmor-frequency RF fields. However, they are able to orient when tested in such RF fields if trained under this condition, indicating that the RF field alters, but does not eliminate, the magnetic input. Larmor-frequency RF fields of higher intensities, with or without harmonics, dramatically alter the magnetic compass response. In contrast, exposure to broadband RF fields in training, in testing, or in both training and testing eliminates magnetic compass information. These findings demonstrate that low-level RF fields at intensities found in many laboratory and field experiments may have very different effects on the perception of the magnetic field in birds, depending on the type and intensity of the RF field, and the birds' familiarity with the RF-generated pattern., (© 2023. The Author(s).)

Many organisms fail to adjust their phenology sufficiently to climate change. Studies have concentrated on adaptive responses within localities, but little is known about how latitudinal dispersal enhances evolutionary potential. Rapid adaptation is expected if dispersers from lower latitudes have improved synchrony to northern conditions, thereby gain fitness and introduce genotypes on which selection acts. Here we provide experimental evidence that dispersal in an avian migrant enables rapid evolutionary adaptation. We translocated Dutch female pied flycatchers (Ficedula hypoleuca) and eggs to Sweden, where breeding phenology is ~15 days later. Translocated females bred earlier, and their fitness was 2.5 times higher than local Swedish flycatchers. We show that between-population variation in timing traits is highly heritable, and hence immigration of southern genotypes promotes the necessary evolutionary response. We conclude that studies on adaptation to large-scale environmental change should not just focus on plasticity and evolution based on standing genetic variation but should also include phenotype-habitat matching through dispersal as a viable route to adjust., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Plumage color has traditionally been regarded as a static ornamental trait, but evidence is accumulating for significant color changes without molt that typically reduce the conspicuousness of ornamentation. In some species, the social partner seems to increase its reproductive investment if the color trait is experimentally enhanced, suggesting that color change could act as a signal. However, the information content of this signal is so far unclear. For example, birds in poor condition or making greater effort may deteriorate more severely. We used brood size manipulations to alter the reproductive effort of male and female collared flycatchers Ficedula albicollis. Both sexes showed less severe decline in some reflectance attribute of their white breast when their brood was experimentally reduced. In each sex, greater deterioration of the reflectance trait affected by the manipulation was accompanied by increased feeding rate by the partner. These feeding patterns do not prove, but are consistent with, a compensatory response by the partner to induced degradation. The manipulation effects on color change we detected confirm for the first time that plumage color deterioration can indicate current reproductive effort, thereby providing a potential fitness advantage to social partners that react to such deterioration., (© 2023. The Author(s).)

The vocal control nucleus HVC in songbirds has emerged as a widespread model system to study adult brain plasticity in response to changes in the hormonal and social environment. I review here studies completed in my laboratory during the last decade that concern two aspects of this plasticity: changes in aggregations of extracellular matrix components surrounding the soma of inhibitory parvalbumin-positive neurons called perineuronal nets (PNN) and the production/incorporation of new neurons. Both features are modulated by the season, age, sex and endocrine status of the birds in correlation with changes in song structure and stability. Causal studies have also investigated the role of PNN and of new neurons in the control of song. Dissolving PNN with chondroitinase sulfate, a specific enzyme applied directly on HVC or depletion of new neurons by focalized X-ray irradiation both affected song structure but the amplitude of changes was limited and deserves further investigations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

How the accelerating pace of global warming will affect animal populations depends on the effects of increasing temperature across the life cycle. Developing young are sensitive to environmental challenges, often with life-long consequences, but the risks of climate warming during this period are insufficiently understood. This may be due to limited insight into physiological sensitivity and the temperatures that represent a thermal challenge for young. Here we examined the physiological and behavioural effects of increasing temperatures by measuring metabolic rate, water loss, and heat dissipation behaviours between 25-45 °C in nestlings of a small free-living songbird of temperate SE-Australia, the superb fairy-wren. We found a high and relatively narrow thermoneutral zone from 33.1 to 42.3 °C, with metabolic rate increasing and all nestlings panting above this range. Evaporative water loss sharply increased above 33.5 °C; at the same temperature, nestlings changed their posture (extended their wings) to facilitate passive heat loss. However, at all temperatures measured, water loss was insufficient to dissipate metabolically produced heat, indicating poor cooling capabilities, which persisted even when individuals were panting. While nestlings are relatively tolerant to higher temperatures, with no evidence for hyperthermia at temperatures below 42 °C, they are at a high risk of dehydration even at lower temperatures, with limited ability to mitigate this. Thus, climate warming is likely to elevate the risk dehydration, which is concerning, since it is accompanied by drier conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Aggression plays a crucial role in deterring predators and securing resources to promote fitness. Nevertheless, studies focussing on female aggression remain scarce. In songbirds, aggression is prevalent during the breeding season, when same-sex individuals compete for limited resources. Additionally, females of some bird species exhibit snake-like hissing behaviour during incubation presumably to lower predation rates and improve fitness. Such behaviours may co-vary, forming a behavioural syndrome that could constrain trait expression. Here, we investigated a resident population of blue tits (Cyanistes caeruleus), to examine the repeatability and covariation of female-female aggression and hissing behaviour, aiming to determine if these constitute a behavioural syndrome. We quantified female-female aggression during simulated territorial intrusions and measured number of hissing calls in response to a simulated predator intrusion into the nest box. We found that both female-female aggression and hissing behaviour were repeatable traits, and that older females approached the intruder less. However, we found no evidence of covariation between female-female aggression and hissing behaviour. Thus, our findings suggest that female-female aggression and hissing behaviour, although both displayed in a nest defence context, are evolutionarily independent traits in the blue tit., (© 2023. Springer Nature Limited.)

Birds have a diverse acoustic communication system, and the ability to recognise their own species' song from a distance facilitates complex behaviours related to mate attraction and rival deterrence. However, certain species, including doves, do not learn songs and their vocal repertoires are much simpler than those of better-studied songbirds. In these so-called non-learning birds, relatively little is known about the role that bird song plays in intra- and interspecific interactions, and how such behaviours might be acquired (inherited or learned from experience). To investigate this question, we focused on two species of African wood doves whose long-range songs are used in a territorial context. Specifically, we examined the responses of sympatric and allopatric populations of male blue-headed wood-doves (Turtur brehmeri) and tambourine doves (Turtur tympanistria) to different types of simulated territorial intrusions, i.e. playback of conspecific, congeneric, and control songs. We aimed to assess (i) whether these species, which have similar songs, respond only to their own species' song or exhibit interspecific territoriality, and (ii) if the response pattern is affected by the presence or absence of congeners in the general area. We found that both species responded strongly to playback of their own species in both sympatric and allopatric populations. In allopatry, though, male tambourine doves misdirected their response and also approached the playback of congeneric songs. Our results indicate that, in areas where the studied Turtur doves live in sympatry, they do not exhibit consistent interspecific territoriality. However, we cannot exclude the possibility that the smaller tambourine dove avoids its larger congener during the process of territory establishment. The difference in tambourine doves' response toward the song of present (sympatric) or absent (allopatric) congeners suggests that the ability to discriminate between songs of similarly singing potential competitors is acquired through earlier interactions and learning. This plasticity in response supports the misdirected aggression hypothesis, which argues that interspecific territorialism emerges as a maladaptive by-product of signal similarity. However, on an evolutionary timescale, such an ability could be considered an adaptive cognitive tool useful for resolving competing interests with congeners., (© 2023. Springer Nature Limited.)

Predation or brood parasitism risks can change the behaviors and reproductive decisions in many parental animals. For oviparous species, mothers can mitigate their reproductive success in at least three ways: (1) by avoiding nest sites with high predation or parasitism risks, (2) through hormonal maternal effects that developmentally prime offspring for survival in risky environments, or (3) by investing less in reproduction when predation or parasitism risks are high. Here, we tested if perceived predation and parasitism risks can induce any of these behavioral or physiological responses by exposing female red-winged blackbirds (Agelaius phoeniceus) to playbacks of two major nest threats, a predator (Cooper's hawk, Accipiter cooperii) and an obligate brood parasite (brown-headed cowbird; Molothrus ater), as well as two controls (harmless Eastern meadowlark, Sturnella magna; and silence). We found that female blackbirds did not avoid nesting at sites treated with predator or brood parasite playbacks, nor were females more likely to abandon nesting attempts at these sites. Egg size and yolk hormone profiles, which are common proxies for maternal investment in oviparous species, were statistically similar across treatment sites. Instead, we found intraclutch variation in yolk steroid hormone profiles: concentrations of three progestogens (pregnanedione, 17α-hydroxypregnenolone, and deoxycorticosterone) and two androgens (testosterone and androstenedione) were higher in third-laid than first-laid eggs. Our study largely confirms previous findings of consistent intraclutch yolk hormone variation in this species, in birds in general, and in other oviparous lineages, but uniquely reports on several yolk steroid hormones largely overlooked in the literature on hormone-mediated maternal effects., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Complex vocal learning, a critical component of human spoken language, has been assumed to be associated with more-advanced cognitive abilities. Tests of this hypothesis between individuals within a species have been inconclusive and have not been done across species. In this work, we measured an array of cognitive skills-namely, problem-solving, associative and reversal learning, and self-control-across 214 individuals of 23 bird species, including 19 wild-caught songbird species, two domesticated songbird species, and two wild-caught vocal nonlearning species. We found that the greater the vocal learning abilities of a species, the better their problem-solving skills and the relatively larger their brains. These conclusions held when controlling for noncognitive variables and phylogeny. Our results support a hypothesis of shared genetic and cognitive mechanisms between vocal learning, problem-solving, and bigger brains in songbirds.

Migratory flight requires birds to maintain intensive aerobic exercise for many hours or days. Maintaining O2 supply to flight muscles is therefore important during migration, especially since migratory songbirds have been documented flying at altitudes greater than 5000 m above sea level, where O2 is limited. Whether songbirds exhibit seasonal plasticity of the O2 cascade to maintain O2 uptake and transport during migratory flight is not well understood. We investigated changes in the hypoxic ventilatory response, haematology and pectoralis (flight) muscle phenotype of 6 songbird species from 3 families during migratory and non-migratory conditions. Songbirds were captured during southbound migration in southern Ontario, Canada. Half of the birds were assessed during migration, and the rest were transitioned onto a winter photoperiod to induce a non-migratory phenotype and measured. All species exhibited seasonal plasticity at various stages along the O2 cascade, but not all species exhibited the same responses. Songbirds tended to be more hypoxia tolerant during migration, withstanding 5 kPa O2 and breathed more effectively through slower, deeper breaths. Warblers had a stronger haemoglobin-O2 affinity during autumn migration (decrease of ∼4.7 Torr), while the opposite was observed in thrushes (increase of ∼2.6 Torr). In the flight muscle there was an ∼1.2-fold increase in the abundance of muscle fibres with smaller fibre transverse areas during autumn migration, but no changes in capillary:fibre ratio. These adjustments would enhance O2 uptake and transport to the flight muscle. Our findings demonstrate that in the O2 cascade there is no ideal migratory phenotype for all songbirds., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

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

A major question in behavioural ecology is why behaviour, physiology and morphology are often integrated into syndromes. In great tits, Parus major, for example, explorative males are larger (vs. smaller) and leaner (vs. heavier) compared to less explorative individuals. Unfortunately, considerable debate exists on whether patterns found in specific studies are replicable. This debate calls for study replication among species, populations and sexes. We measured behavioural (exploration), physiological (breathing rate) and morphological traits (body mass, tarsus length, wing length, bill length) in two species (great vs. blue tits Cyanistes caeruleus), two populations (Forstenrieder Park vs. Starnberg) and two sexes (males vs. females). We then tested whether the same pattern of integration characterized all unique combinations of these three biological categories (hereafter called datasets). We used a multi-year repeated measures set-up to estimate among-individual trait correlation matrices for each dataset. We then used structural equation modelling to test for size-dependent behaviour and physiology, size-corrected (i.e. size-independent) behaviour-physiology correlations and size-corrected body mass-dependent behaviour and physiology. Finally, we used meta-analyses to test which structural paths were generally (vs. conditionally) supported (vs. unsupported). We found general and consistent support for size-dependent physiology and size-corrected body mass-dependent physiology across datasets: faster breathers were smaller but heavier for their size. Unexpectedly, condition-dependent behaviour was not supported: explorative birds were neither leaner, nor was this relationship heterogeneous across datasets. All other hypothesized patterns were dataset-specific: the covariance between size and behaviour, and between behaviour and physiology differed in sign between datasets, and both were, on average, not supported. This heterogeneity was not explained by any of our moderators: species, population or sex. The specific pattern of size- and condition-dependent physiology reported for a unique combination of species, population, and sex, thus generally predicted those in others. Patterns of size- or condition-dependent behaviour (i.e. 'personality'), or behaviour-physiology syndromes reported in specific datasets, by contrast, did not. These findings call for studies revealing the ecological background of this variation and highlight the value of study replication to help understand whether patterns of phenotypic integration reported in one study can be generalized., (© 2023 The Authors. Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.)

Young animals need to grow to a large body size fast to maximise their survival prospects until sexual maturity. However, body size varies substantially in wild populations, and neither the selection pressures maintaining this variation, nor the regulatory mechanisms are well understood. IGF-1 administration has been shown to accelerate growth, but this does not necessarily imply that natural variation in growth rate is IGF-1 dependent. To test the latter we administered OSI-906 to pied flycatcher Ficedula hypoleuca nestlings, which has an inhibitory effect on IGF-1 receptor activity. We performed the experiment in two breeding seasons to test the prediction that blocking the IGF-1 receptor downregulates growth. As predicted, OSI-906 treated nestlings had lower body mass and reached a smaller structural size than siblings receiving a vehicle only, with the mass difference being most profound at the age preceding the highest body mass growth rate. The IGF-1 receptor inhibition effect on growth varied with age and year of study, and we discuss possible explanations. The OSI-906 administration results indicate that natural variation in growth rate is regulated by IGF-1, and constitutes a novel tool to study causes and consequences of growth variation, but details of the underlying mechanism still need to be resolved., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

While Plasmodium parasitism is common in songbirds, its impact on avian reproduction is unclear owing to conflicting reports in the existing literature. Particularly understudied is the impact of phase of infection on variation in host reproductive physiology in wild, breeding-condition birds. However, assessing the full impact of Plasmodium on reproductive success in the wild can be difficult because individuals experiencing severe effects of parasitism may not enter the breeding population and may be less likely to be captured during field studies. To address these factors, we quantified metrics of health and reproductive physiology in wild-caught, breeding-condition male dark-eyed juncos (Junco hyemalis hyemalis) before and after experimental Plasmodium inoculation in a captive setting. Metrics of health and reproductive physiology included activity rate, hematocrit, scaled body mass, testosterone and sperm production. Individuals already infected at capture (i.e., chronically infected) had higher levels of hematocrit than males without chronic infections. Experimentally infected males showed a larger reduction in hematocrit and activity rate as compared to controls. However, chronic infection status did not influence the extent of metric decline. Testosterone production did not vary by treatment and most birds produced sperm following inoculation. Broadly, our results suggest that male juncos exposed to Plasmodium during the breeding season likely experience declines in general health, but Plasmodium infections do not negatively impact reproductive physiology. We conclude that physiological tradeoffs in males may favor maintenance of reproductive function despite infection., (© 2023. Springer Nature Limited.)

Anthropogenic stressors, such as artificial light at night (ALAN), increasingly affect the sleep behaviour and physiology of wild birds, particularly in areas where human activity is prevalent. To understand the consequences of the resulting sleep deprivation, it is essential to investigate whether the effects of sleep deprivation on cognitive performance, observed in humans, also occur in birds. Here, we studied the impact of sleep deprivation, induced by intermittent exposure to ALAN, on inhibitory control, vigilance behaviour, and exploratory behaviour in great tits. Furthermore, we hypothesised that the effect of ALAN could depend on an individual's natural sleep duration and the timing of sleep. To achieve these goals, we measured emergence and entry times from/into the nest box in the wild, before capturing the great tits. In captivity, half of the birds were exposed to intermittent ALAN, and cognitive performance was assessed the following morning for all birds. ALAN-exposed birds were less successful on the detour reach task and when they started pecking at the test tube, they pecked more often. However, neither of the effects was related to the natural sleep duration or timing, in contrast to our hypothesis, and there were no differences between the ALAN-exposed and non-exposed group in vigilance and exploratory behaviour. Thus, even one night of exposure to ALAN can negatively affect cognitive performance in wild birds, possibly with negative effects on their performance and survival., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Interactive vocal communication, similar to a human conversation, requires flexible and real-time changes to vocal output in relation to preceding auditory stimuli. These vocal adjustments are essential to ensuring both the suitable timing and content of the interaction. Precise timing of dyadic vocal exchanges has been investigated in a variety of species, including humans. In contrast, the ability of non-human animals to accurately adjust specific spectral features of vocalization extemporaneously in response to incoming auditory information is less well studied. One spectral feature of acoustic signals is the fundamental frequency, which we perceive as pitch. Many animal species can discriminate between sound frequencies, but real-time detection and reproduction of an arbitrary pitch have only been observed in humans. Here, we show that nightingales in the wild can match the pitch of whistle songs while singing in response to conspecifics or pitch-controlled whistle playbacks. Nightingales matched whistles across their entire pitch production range indicating that they can flexibly tune their vocal output along a wide continuum. Prompt whistle pitch matches were more precise than delayed ones, suggesting the direct mapping of auditory information onto a motor command to achieve online vocal replication of a heard pitch. Although nightingales' songs follow annual cycles of crystallization and deterioration depending on breeding status, the observed pitch-matching behavior is present year-round, suggesting a stable neural circuit independent of seasonal changes in physiology. Our findings represent the first case of non-human instantaneous vocal imitation of pitch, highlighting a promising model for understanding sensorimotor transformation within an interactive context. VIDEO ABSTRACT., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Migratory songbirds have the remarkable ability to extract directional information from the Earth's magnetic field 1,2 . The exact mechanism of this light-dependent magnetic compass sense, however, is not fully understood. The most promising hypothesis focuses on the quantum spin dynamics of transient radical pairs formed in cryptochrome proteins in the retina 3-5 . Frustratingly, much of the supporting evidence for this theory is circumstantial, largely because of the extreme challenges posed by genetic modification of wild birds. Drosophila has therefore been recruited as a model organism, and several influential reports of cryptochrome-mediated magnetic field effects on fly behaviour have been widely interpreted as support for a radical pair-based mechanism in birds 6-23 . Here we report the results of an extensive study testing magnetic field effects on 97,658 flies moving in a two-arm maze and on 10,960 flies performing the spontaneous escape behaviour known as negative geotaxis. Under meticulously controlled conditions and with vast sample sizes, we have been unable to find evidence for magnetically sensitive behaviour in Drosophila. Moreover, after reassessment of the statistical approaches and sample sizes used in the studies that we tried to replicate, we suggest that many-if not all-of the original results were false positives. Our findings therefore cast considerable doubt on the existence of magnetic sensing in Drosophila and thus strongly suggest that night-migratory songbirds remain the organism of choice for elucidating the mechanism of light-dependent magnetoreception., (© 2023. The Author(s).)

Hybrid zones can be used to identify traits that maintain reproductive isolation and contribute to speciation. Cognitive traits may serve as post-mating reproductive isolating barriers, reducing the fitness of hybrids if, for example, misexpression occurs in hybrids and disrupts important neurological mechanisms. We tested this hypothesis in a hybrid zone between two subspecies of Swainson's thrushes (Catharus ustulatus) using two cognitive tests-an associative learning spatial test and neophobia test. We included comparisons across the sexes and seasons (spring migration and winter), testing if hybrid females performed worse than males (as per Haldane's rule) and if birds (regardless of ancestry or sex) performed better during migration, when they are building navigational maps and encountering new environments. We documented reduced cognitive abilities in hybrids, but this result was limited to males and winter. Hybrid females did not perform worse than males in either season. Although season was a significant predictor of performance, contrary to our prediction, all birds learned faster during the winter. The hypothesis that cognitive traits could serve as post-mating isolating barriers is relatively new; this is one of the first tests in a natural hybrid zone and non-food-caching species. We also provide one of the first comparisons of cognitive abilities between seasons. Future neurostructural and neurophysiological work should be used to examine mechanisms underlying our behavioral observations., (© 2023. The Author(s).)

Hosts of obligate avian brood parasites often evolve defense mechanisms to avoid rearing unrelated young. One common defense is egg rejection, for which hosts often rely on eggshell color. Most research has assumed that hosts respond to perceived color differences between their own eggs and parasite eggs regardless of the particular color; however, recent experiments have found that many hosts respond more strongly to brown foreign eggs than to equally dissimilar blue eggs. Yet, none of these prior studies tested a brown-egg-laying species and, with only one exception, all were conducted in open nests where light levels are considered sufficient for effective color-based egg discrimination. Here, we explored how two cavity-nesting hosts of the parasitic brown-headed cowbird (Molothrus ater) - the blue-egg-laying eastern bluebird (Sialia sialis) and the brown-egg-laying house wren (Troglodytes aedon) - respond to experimental eggs painted six distinct colors ranging from blue to brown. Rejection responses of both hosts were best predicted by perceived differences in color between the model egg and their own eggs. Specifically, we found that house wrens preferentially rejected eggs bluer than their own eggs. However, although we found that bluebirds relied on perceived differences in color for their egg rejection decisions, further tests are needed to determine whether they preferentially rejected brown eggs or simply responded to absolute perceived differences in color. These findings demonstrate that these cavity-nesting birds treat perceived color differences in distinct ways, which has important implications on the coevolutionary arms races and the interpretation of avian-perceived color differences., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Lead exposure is a concern in urban ecosystems, with physiological and behavioral effects well documented in humans. Wildlife inhabiting urban ecosystems are also exposed to lead, yet little work has documented the sublethal effects of lead exposure in urban wildlife. We studied northern mockingbirds (Mimus polyglottos) in three neighborhoods of New Orleans, Louisiana, two with high soil lead and one with low soil lead, to better understand how lead exposure may influence mockingbirds' reproductive biology. We monitored nesting attempts, measured lead concentrations in blood and feathers of nestling mockingbirds, documented egg hatching and nesting success, and assessed rates of sexual promiscuity in relation to neighborhood soil lead levels. We found that nestling mockingbirds' blood and feather lead levels reflected the soil lead levels of their neighborhoods and nestling blood lead levels were similar to those of adult mockingbirds in the same neighborhoods. Nest success, as evaluated by daily nest survival rates, was higher in the lower lead neighborhood. Clutch sizes varied substantially across neighborhoods, but rates of unhatched eggs did not covary with neighborhood lead levels, suggesting that other drivers are influencing variation in clutch sizes and hatching success in urban habitats. At least one-third of nestling mockingbirds were sired by an extra-pair male, and there was no relationship between extra-pair paternity rates and neighborhood lead levels. This study provides insight on how lead contamination may influence reproduction in urban-dwelling wildlife and suggests that nestling birds could serve as useful bioindicators of lead levels in urban neighborhoods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Sensory feedback is required for the stable execution of learned motor skills, and its loss can severely disrupt motor performance. The neural mechanisms that mediate sensorimotor stability have been extensively studied at systems and physiological levels, yet relatively little is known about how disruptions to sensory input alter the molecular properties of associated motor systems. Songbird courtship song, a model for skilled behavior, is a learned and highly structured vocalization that is destabilized following deafening. Here, we sought to determine how the loss of auditory feedback modifies gene expression and its coordination across the birdsong sensorimotor circuit. To facilitate this system-wide analysis of transcriptional responses, we developed a gene expression profiling approach that enables the construction of hundreds of spatially-defined RNA-sequencing libraries. Using this method, we found that deafening preferentially alters gene expression across birdsong neural circuitry relative to surrounding areas, particularly in premotor and striatal regions. Genes with altered expression are associated with synaptic transmission, neuronal spines, and neuromodulation and show a bias toward expression in glutamatergic neurons and Pvalb/Sst- class GABAergic interneurons. We also found that connected song regions exhibit correlations in gene expression that were reduced in deafened birds relative to hearing birds, suggesting that song destabilization alters the inter-region coordination of transcriptional states. Finally, lesioning LMAN, a forebrain afferent of RA required for deafening-induced song plasticity, had the largest effect on groups of genes that were also most affected by deafening. Combined, this integrated transcriptomics analysis demonstrates that the loss of peripheral sensory input drives a distributed gene expression response throughout associated sensorimotor neural circuitry and identifies specific candidate molecular and cellular mechanisms that support the stability and plasticity of learned motor skills., Competing Interests: BC, KL, RV, MB No competing interests declared, FG Foad Green is affiliated with Syapse, Inc. The author has no financial interests to declare, (© 2023, Colquitt et al.)

Neuronal tuning for spectral and temporal features has been studied extensively in the auditory system. In the auditory cortex, diverse combinations of spectral and temporal tuning have been found, but how specific feature tuning contributes to the perception of complex sounds remains unclear. Neurons in the avian auditory cortex are spatially organized in terms of spectral or temporal tuning widths, providing an opportunity for investigating the link between auditory tuning and perception. Here, using naturalistic conspecific vocalizations, we asked whether subregions of the auditory cortex that are tuned for broadband sounds are more important for discriminating tempo than pitch, due to the lower frequency selectivity. We found that bilateral inactivation of the broadband region impairs performance on both tempo and pitch discrimination. Our results do not support the hypothesis that the lateral, more broadband subregion of the songbird auditory cortex contributes more to processing temporal than spectral information., (© 2023. The Author(s).)

Addressing the patterns of variation in thermal traits is crucial to better predict the potential effects of climate change on organisms. Here, we assessed seasonal (winter vs summer) adjustments in key thermoregulatory traits in eight Mediterranean-resident songbirds. Overall, songbirds increased whole-animal (by 8%) and mass-adjusted (by 9%) basal metabolic rate and decreased (by 56%) thermal conductance below the thermoneutral zone during winter. The magnitude of these changes was within the lower values found in songbirds from northern temperate areas. Moreover, songbirds increased (by 11%) evaporative water loss within the thermoneutral zone during summer, while its rate of increase above the inflection point of evaporative water loss (i.e., the slope of evaporative water loss versus temperature) decreased by 35% during summer - a value well above that reported for other temperate and tropical songbirds. Finally, body mass increased by 5% during winter, a pattern similar to that found in many northern temperate species. Our findings support the idea that physiological adjustments might enhance the resilience of Mediterranean songbirds to environmental changes, with short-term benefits by saving energy and water under thermally stressful conditions. Nevertheless, not all species showed the same patterns, suggesting different strategies in their thermoregulatory adaptations to seasonal environments., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest to declare., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

The transcriptional regulation of innate immune function across annual life history states (LHS) remains obscure in avian migrants. We, therefore, investigated this in a migratory passerine songbird, redheaded bunting (Emberiza bruniceps), which exhibits long-distance vernal migration from India to Central Asia. We exposed the birds (N = 10) to differential photoperiodic conditions to induce a non-migratory (NM), pre-migratory (PM), migratory (MIG), and refractory (REF) state, and performed gene expression assays of melatonin receptors (MEL1A and MEL1B), and innate immunity-linked genes (IL1B, IL6, TLR4, and NFKB) in spleen and blood. We found a significant reduction in splenic mass and volume, and a parallel increase in fat accumulation, and testicular growth in birds under migratory state. The gene expression assay revealed an upregulation of MEL1A and MEL1B mRNA levels in both the tissues in MIG. Additionally, we found a nocturnal increase of splenic IL1B expression, and IL1B, IL6, and TLR4 expression in the blood. The mRNA expression of melatonin receptors and proinflammatory cytokine showed a positive correlation. These results suggest that melatonin relays the photoperiodic signal to peripheral immune organs, which shows LHS-dependent changes in mRNA expression of immune genes., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests. The following are the supplementary data related to this article. Supplementary data to this article can be found online at https://doi.org/10.1016/j.cbpa.2023.111381., (Copyright © 2023 Elsevier Inc. All rights reserved.)

AbstractEnvironmental challenges faced early in life can both activate and shape the development of the hypothalamic-pituitary-adrenal axis. Activation of this axis is characterized in part by elevated levels of glucocorticoids, exposure to which can have profound effects throughout an animal's life. We have demonstrated that in nestling eastern bluebirds ( Sialia sialis ), bouts of environmentally relevant cooling result in elevations of corticosterone (the primary avian glucocorticoid) very early in life. Nestlings repeatedly exposed to cooling also exhibit dampened corticosterone secretion later in life in response to restraint compared to control nestlings. We explored the mechanistic basis of this phenomenon. Specifically, we asked whether early-life cooling alters adrenal sensitivity to adrenocorticotropic hormone (ACTH), the primary controller of corticosterone synthesis and release. To this end, we subjected nestlings to repeated bouts of cooling (cooled nestlings) or brooding temperatures (control nestlings) early in life and, before fledging, assessed (1) the capacity of the nestlings' adrenals to produce corticosterone following ACTH injection, (2) the effect of cooling on corticosterone responses to restraint, and (3) the effect of cooling on adrenal sensitivity to ACTH. We found that both cooled and control nestlings secreted substantially higher levels of corticosterone following ACTH treatment than they did following restraint. We also confirmed that cooled nestlings had reduced corticosterone secretion in response to restraint compared to control nestlings; however, sensitivity to exogenous ACTH did not differ between temperature treatments. We hypothesize that early-life cooling alters later corticosterone secretion by affecting higher levels of the hypothalamic-pituitary-adrenal axis.

During migration, long-distance migratory songbirds may fly nonstop for days, whereas shorter-distance migrants complete flights of 6 to 10 h. Fat is the primary fuel source, but protein is also assumed to provide a low, consistent amount of energy for flight. However, little is known about how the use of these fuel sources differs among bird species and in response to flight duration. Current models predict that birds can fly until fat stores are exhausted, with little consideration of protein's limits on flight range or duration. We captured two related migratory species-ultra long-distance blackpoll warblers ( Setophaga striata ) and short-distance yellow-rumped warblers ( Setophaga coronata )-during fall migration and flew them in a wind tunnel to examine differences in energy expenditure, overall fuel use, and fuel mixture. We measured fat and fat-free body mass before and after flight using quantitative magnetic resonance and calculated energy expenditure from body composition changes and doubly labeled water. Three blackpolls flew voluntarily for up to 28 h-the longest wind tunnel flight to date-and ended flights with substantial fat reserves but concave flight muscle, indicating that protein loss, rather than fat, may actually limit flight duration. Interestingly, while blackpolls had significantly lower mass-specific metabolic power in flight than that of yellow-rumped warblers and fuel use was remarkably similar in both species, with consistent fat use but exceptionally high rates of protein loss at the start of flight that declined exponentially over time. This suggests that protein may be a critical, dynamic, and often overlooked fuel for long-distance migratory birds.

Multilevel societies are formed when stable groups of individuals spatially overlap and associate preferentially with other groups, producing a hierarchical social structure. 1 Once thought to be exclusive to humans and large mammals, these complex societies have recently been described in birds. 2 , 3 However, it remains largely unclear what benefits individuals gain by forming multilevel societies. 1 One hypothesis-based on food sharing in hunter-gatherers 4 -is that multilevel societies facilitate access to a range of cooperative relationships, with individual investment varying across the hierarchical levels of the society. We tested experimentally whether such graded cooperation occurs in the multilevel society of a songbird, the superb fairy-wren (Malurus cyaneus). Specifically, we measured whether responses to playbacks of distress calls-used to recruit help when in extreme danger-varied according to the social level at which the focal individual is connected with the caller. We predicted that anti-predator responses should be highest within breeding groups (the core social unit), intermediate between groups from the same community, and lowest across groups from different communities. Our results confirm that birds exhibit the predicted hierarchical pattern of helping and that, within breeding groups, this pattern is independent of kinship. This pattern of graded helping responses supports the hypothesis that multilevel social structures can sustain stratified cooperative relationships and reveals similarity in cooperation in qualitatively different behaviors-anti-predator behavior and food sharing-in the multilevel societies of songbirds and humans., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Golden-winged Warblers (Vermivora chrysoptera, Parulidae) are declining migrant songbirds that breed in the Great Lakes and Appalachian regions of North America. Within their breeding range, Golden-winged Warblers are found in early successional habitats adjacent to mature hardwood forest, and previous work has found that Golden-winged Warbler habitat preferences are scale-dependent. Golden-winged Warbler Working Group management recommendations were written to apply to large regions of the breeding range, but there may be localized differences in both habitat availability and preferences. Rapid declines at the southernmost extent of their breeding range in Western North Carolina necessitate investigation into landscape characteristics governing distribution in this subregion. Furthermore, with the increase in availability of community science data from platforms such as eBird, it would be valuable to know if community science data produces similar distribution models as systemic sampling data. In this study, we described patterns of Golden-winged Warbler presence in Western North Carolina by examining habitat variables at multiple spatial scales using data from standardized Audubon North Carolina (NC) playback surveys and community science data from eBird. We compared model performance and predictions between Audubon NC and eBird models and found that Golden-winged Warbler presence is associated with sites which, at a local scale (150m), have less mature forest, more young forest, more herb/shrub cover, and more road cover, and at a landscape scale (2500m), have less herb/shrub cover. Golden-winged Warbler presence is also associated with higher elevations and smaller slopes. eBird and Audubon models had similar variable importance values, response curves, and overall performance. Based on variable importance values, elevation, mature forest at the local scale, and road cover at the local scale are the primary variables driving the difference between Golden-winged Warbler breeding sites and random background sites in Western North Carolina. Additionally, our results validate the use of eBird data, since they produce species distribution modeling results that are similar to results obtained from more standardized survey methods., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Whitenack et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

North American grassland birds have widely declined over the past 50 years, largely due to anthropogenic-driven loss of native prairie habitat. In response to these declines, many conservation programs have been implemented to help secure wildlife habitat on private and public lands. The Grasslands Coalition is one such initiative established to advance the conservation of grassland birds in Missouri. The Missouri Department of Conservation conducted annual point count surveys for comparison of grassland bird relative abundance between focal grassland areas and nearby paired (i.e., containing no targeted management) sites. We analyzed 17 years of point count data with a generalized linear mixed model in a Bayesian framework to estimate relative abundance and trends across focal or paired sites for nine bird species of management interest that rely on grasslands: barn swallow (Hirundo rustica), brown-headed cowbird (Molothrus ater), dickcissel (Spiza americana), eastern meadowlark (Sturnella magna), grasshopper sparrow (Ammodramus savannarum), Henslow's sparrow (A. henslowii), horned lark (Eremophila alpestris), northern bobwhite (Colinus virginianus), and red-winged blackbird (Agelaius phoeniceus). Relative abundance of all species except eastern meadowlarks declined regionally. Relative abundance of barn swallows, brown-headed cowbirds, dickcissels, eastern meadowlarks, Henslow's sparrows, and northern bobwhites was higher in focal than paired sites, though relative abundance trends were only improved in focal vs. paired areas for dickcissels and Henslow's sparrows. Relative abundance increased with increasing grassland cover at the local (250-m radius) scale for all species except horned larks and red-winged blackbirds and at the landscape (2,500-m radius) scale for all species except dickcissels, eastern meadowlarks, and northern bobwhites. Our results suggest focal areas contained greater relative abundances of several grassland species of concern, likely due to increased availability of grassland habitat at local and landscape scales. Further efforts to decrease landscape-scale fragmentation and improve habitat quality may be needed to achieve conservation goals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Disentangling the interaction between the genetic basis and environmental context underlying phenotypic variation is critical for understanding organismal evolution. Environmental change, such as increased rates of urbanization, can induce shifts in phenotypic plasticity with some individuals adapting to city life while others are displaced. A key trait that can facilitate adaptation is the degree at which animals respond to stressors. This stress response, which includes elevation of baseline circulating concentrations of glucocorticoids, has a heritable component and exhibits intra- and inter-individual variation. However, the mechanisms behind this variability and whether they might be responsible for adaptation to different environments are not known. Variation in DNA methylation can be a potential mechanism that mediates environmental effects on the stress response, as early-life stressors increase glucocorticoid concentrations and change adult phenotype. We used an inter- and intra-environmental cross-foster experiment to analyse the contribution of DNA methylation to early-life phenotypic variation. We found that at hatching, urban house wren (Troglodytes aedon) offspring had higher methylation frequencies compared with their rural counterparts. We also observed age-related patterns in offspring methylation, indicating the developmental effects of the rearing environment on methylation. At fledgling, differential methylation analyses showed that cellular respiration genes were differentially methylated in broods of different origins and behavioural and metabolism genes were differentially methylated in broods of different rearing environments. Lastly, hyper-methylation of a single gene (CNTNAP2) is associated with decreased glucocorticoid levels and the rearing environment. These differential methylation patterns linked to a specific physiological phenotype suggest that DNA methylation may be a mechanism by which individuals adjust to novel environments during their lifespan. Characterizing genetic and environmental influences on methylation is critical for understanding the role of epigenetic mechanisms in evolutionary adaptation., (© 2022 The Authors. Journal of Animal Ecology © 2022 British Ecological Society.)