Urbanization drastically alters environments, presenting wildlife with novel challenges to which they must adjust to persist. Often, an animal's first response to environmental change is to shift behavior, which requires processing environmental cues into neural signals that can govern adaptive behavior. However, the neural mechanisms underpinning behavioral responses to urbanization are largely unresolved. During the breeding season, urban songbirds of several species are more aggressive than their rural counterparts. Among the many neural mechanisms that regulate aspects of aggression, arginine vasotocin (AVT) is a neuropeptide highly implicated in the regulation of sociality, including territorial aggression. Prior work has shown that the abundance of AVT peptide is associated with behavioral responses to urbanization. However, social behavior is also influenced by AVT receptor availability, making it important to compare receptor distribution and expression between urban and rural birds. To address this gap in understanding of how AVT receptors could contribute to behavioral responses to urbanization, we used in situ hybridization to describe the distribution of one AVT receptor known to influence behavior, AVT3R, throughout the brain of adult urban and rural male song sparrows (Melospiza melodia). Additionally, we used quantitative PCR (qPCR) to compare the relative expression of mRNA for AVTI1 (AVT), and two of its receptors (AVT3R and AVT4R) in the hypothalamus. We predicted that urban males would have a higher abundance of AVT3R transcripts in the lateral septum, greater mRNA expression of AVT receptors in the hypothalamus, and a positive correlation of AVT gene targets with aggression compared to rural males. We observed high AVT3R mRNA signal in the lateral septum, medial bed nucleus of the stria terminalis, hippocampus, nidopallium, cerebellum and optic tectum, with diffuse signal in the lateral hypothalamus and nucleus rotundus. There were no significant differences in the number of AVT3R transcripts in the lateral septum between urban and rural males. When we quantified relative mRNA expression of AVT and two of its receptors using qPCR, we found that urban males had higher relative mRNA expression of AVT3R in the hypothalamus compared to rural males, and aggression immediately before capture was negatively associated with the expression of AVT, AVT3R, and AVT4R. Thus, expression of AVT and its receptors has the potential to contribute to higher aggression in male song sparrows but may not explain habitat-related variation.

Urbanization is projected to grow rapidly in the coming decades. Several consequences of this form of anthropogenic change for ecosystems and individuals are described by the fields of urban ecology and urban ecophysiology, respectively. Of the environmental variables that could pose challenges for urban-living animals, energy limitations may be especially important, particularly for animals with critical periods of growth, such as nestling songbirds. Many studies document reduced arthropod prey availability for songbirds living in urban habitats, which can compromise the growth and condition of nestlings, possibly because of protein limitation. In our study system of replicate urban and rural song sparrow (Melospiza melodia) populations, there is lower arthropod biomass in urban habitats, as well as higher rates of brood parasitism by brown-headed cowbirds (Molothrus ater), which can cause additional nutritional limitation. Therefore, we hypothesized that urban nestling song sparrows may be protein-limited and experience compromised growth and body condition relative to rural nestlings. We tested this hypothesis with two studies. First, we took body measurements during the first 10 days of development from rural nestlings, urban nestlings without brood parasites, and urban nestlings with brood parasites to see if groups differed in growth (n = 348 nestlings over 6 years). Second, we collected nestling plasma samples (n = 43) from a subset of the birds measured in the first study and then used liquid chromatography-mass spectrometry (LC-MS) to determine concentrations of free amino acids. We found rural nestlings had greater change over time in body mass and tarsus length compared to both urban groups. However, we found no differences in amino acid concentrations across these nestling groups. Thus, urbanization appears to affect growth rates in song sparrows, especially when brood parasites are present in a nest, but we found no evidence supporting amino acids as the mechanistic link for these differences in our system.

Urbanization represents a dramatic and relatively rapid change in the environment that has profound impacts on wild animals. Shifts in behavior and endocrine mechanisms of stress response could allow animals to successfully survive and reproduce in urban habitats. Numerous studies have examined the behavioral and physiological responses of territory-holding male songbirds to urbanization. However, breeding females likely experience anthropogenic noise, light at night, and human disturbance more frequently, and their behavioral coping responses to these disturbances are limited during incubation. Moreover, breeding females face higher energetic demands (allostatic load). Understanding how some species cope with novel urban habitats requires studying individuals facing the greatest challenges, such as breeding females. Therefore, we compared the glucocorticoid stress response and behavioral recovery from a disturbance between urban and rural female song sparrows (Melospiza melodia) during incubation. If facultative adjustments to the glucocorticoid stress response allow birds to cope with urban habitats, we predicted that urban females would return to parental care behaviors after a standardized stressor as soon or sooner than rural females, and that urban females would have a lower glucocorticoid response to the stressor. We captured female song sparrows at the end of the incubation period and measured their glucocorticoid (corticosterone) levels at baseline and after 30 min of standardized restraint. Concurrently, we installed radio frequency identification (RFID) systems at the nest to capture the time to return to parental care behaviors. We found that incubating urban females had significantly lower corticosterone levels when controlling for sampling timepoint (baseline and restraint-induced) compared to rural. Nest return times did not differ across habitats, and latency to return was not significantly correlated with corticosterone levels. Our findings are consistent with prior work in breeding male song sparrows at our study sites; urban males provide higher parental care and have lower restraint-induced corticosterone levels. The absence of a relationship between glucocorticoids and behavior makes it unlikely that these hormones directly regulate parental care, but lower corticosterone levels in urban birds could reflect stress resistance, which has been hypothesized to permit animals to breed in challenging or novel conditions such as urban habitats.

Biparental care is common in socially monogamous avian species, but both partners may seek extra-pair copulations (EPCs). The relative costs and benefits of EPCs between the sexes are likely complex, yet the implications of EPCs for parental care behavior have been examined predominantly in males. Not only could females benefit from EPCs, but females would have additional information about the likelihood of extra-pair young (EPY) in their nest not available to their partners, which likely influences female behavior. We examined how the presence and abundance of EPY in a nest affect parental behavior in a socially monogamous songbird, song sparrows (Melospiza melodia ). We predicted that females who mated outside the social pair would invest more in a clutch with a higher probability of EPY. We monitored nest visitation rates by male and female social partners as a proxy for parental investment and quantified extra-pair paternity in 45 nests. Maternal visitation rates were higher in nests with EPY compared to nests without, while males did not adjust their investment in relation to the presence of EPY. These findings support our prediction that females who participated in EPC would invest more in the resulting offspring.

Urbanization reflects a major form of environmental change impacting wild birds globally. Whereas urban habitats may provide increased availability of water, some food items, and reduced predation levels compared to rural, they can also present novel stressors including increased light at night, ambient noise, and reduced nutrient availability. Urbanization can also alter levels of brood parasitism, with some host species experiencing elevated levels of brood parasitism in urban areas compared to rural areas. Though the demographic and behavioral consequences of urbanization and brood parasitism have received considerable attention, their consequences for cellular-level processes are less understood. Telomeres provide an opportunity to understand the cellular consequences of different environments as they are a well-established metric of biological state that can be associated with residual lifespan, disease risk, and behavior, and are known to be sensitive to environmental conditions. Here we examine the relationships between urbanization, brood parasitism, and blood telomere lengths in adult and nestling song sparrows Melospiza melodia. Song sparrows are a North American songbird found in both urban and rural habitats that experience high rates of brood parasitism by brown-headed cowbirds Molothrus ater in the urban, but not the rural, sites in our study system. Among adults and nestlings from non-parasitized nests, we found no differences in relative telomere lengths between urban and rural habitats. However, among urban nestlings, the presence of a brood parasite in the nest was associated with significantly shorter relative telomere lengths compared to when a brood parasite was absent. Our results suggest a novel, indirect, impact of urbanization on nestling songbirds through the physiological impacts of brood parasitism.

Anthropogenic land use, including urbanization, has caused population declines across diverse taxa including arthropods and songbirds. Declines in one taxa can impact other groups based on its role in a community. In particular, declines in lower trophic level taxa, such as arthropods, could have negative impacts on higher trophic level species. Here, we examined how urban arthropod communities compare to rural ones and how these differences may impact song sparrow (Melospiza melodia) nestlings living in urban and rural habitats. We quantified differences in the abundance, biomass, and diversity of arthropod communities between replicate urban and rural sites. At the same sites, we also compared the stomach contents of nestlings because song sparrows rely upon arthropod prey during development. We found that the arthropod community in urban habitats had lower average abundance, average biomass, and Simpson's diversity compared to rural habitats. The arthropod communities also significantly differed in the relative abundance of some higher trophic level taxa, such as spiders. However, we found no difference in the total stomach content mass, nor the mass of invertebrate food items in the stomachs of urban and rural nestlings. Thus, though urban habitats had lower availability of arthropods, possibly driven by the simplification of urban habitats, there was no evidence of a negative impact the quantity of food provided to urban song sparrow nestlings.

When living in urban habitats, ‘urban adapter’ species often show greater aggression toward conspecifics, yet we do not understand the mechanisms underlying this behavioral shift. The neuroendocrine system regulates socio-sexual behaviors including aggression and thus could mediate behavioral responses to urbanization. Indeed, urban male song sparrows (Melospiza melodia), which are more territorially aggressive, also have greater abundance of the neuropeptide arginine vasotocin (AVT) in nodes of the brain social behavior network. Higher abundance of AVT could reflect long-term synthesis that underlies baseline territoriality or short-term changes that regulate aggression in response to social challenge. To begin to resolve the timeframe over which the AVT system contributes to habitat differences in aggression we used immediate early gene co-expression as a measure of the activation of AVT neurons. We compared Fos induction in AVT-immunoreactive neurons of the bed nucleus of the stria terminalis (BSTm) and paraventricular nucleus of the hypothalamus (PVN) between urban and rural male song sparrows in response to a short (< 5 min.) or long (> 30 min.) song playback to simulate territorial intrusion by another male. We found that urban males had a higher proportion of Fos-positive AVT neurons in both brain regions compared to rural males, regardless of the duration of song playback. Our results suggest that AVT neurons remain activated in urban males, independently of the duration of social challenge. These findings that Fos induction in AVT neurons differs between rural and urban male song sparrows further implicate this system in regulating behavioral responses to urbanization.

Behavioral traits are often the first response to changing environmental conditions, including human induced rapid environmental change. For example, animals living in urban areas are often more aggressive than rural animals. This is especially evident in songbirds; males of several species display elevated aggression in urban habitats. Increased male aggression has been associated with reduced parental care, but the consequences of this trade-off for males, social partners, and offspring in the context of urbanization remains unclear. We explored the effects of increased urban male aggression on the life history traits, parental care, and offspring outcomes of song sparrows (Melospiza melodia). We predicted that urban males would reduce paternal investment and result in urban females providing greater nestling care or reduced fledging success in urban habitats compared to rural. Contrary to our prediction, aggressive urban males did not decrease care but visited the nest more often compared to rural males. Additionally, urban birds had higher nest and fledging success compared to rural, though this was largely due to higher nest predation in rural habitats. Our study is among the first to evaluate trade-offs associated with elevated aggression expressed by urban animals and adds to a growing body of evidence that urban habitats provide benefits to some species.

In many taxa, melanin-based coloration is a signal of dominance or fighting ability and is associated with concentrations of hormones that may mediate aggressive behavior. Previous studies found that experimental manipulation of melanin-based signals can result in manipulated individuals receiving more social challenges in some but not all species. These differences could arise from mismatches between the signal, behavior, and hormone concentrations. In the present study, we experimentally manipulated the chest spotting of urban and rural male song sparrows ( Melospiza melodia ) following an assessment of their territorial aggression and initial concentrations of corticosterone and testosterone and then assessed their behavior and hormone concentrations 2 weeks later. We found that males generally displayed less territorial aggression in the second trial, consistent with our previous findings. Males in the enlarged treatment decreased aggression to a greater degree than those in the reduced treatment. The effect of the plumage manipulation was similar across the rural and urban habitats. Despite the changes in behavior we detected, we found no effects of the manipulation on concentrations of testosterone or corticosterone. Our results show that melanin-based spotting in male song sparrows is a signal of territorial aggression but the physiological mechanisms that mediate the relationships between chest spotting and behavior remain to be identified. Significance statement Many bird species use their plumage to signal their dominance status, fighting ability, or motivation during interactions with other individuals to resolve conflicts without a fight. Here, we asked whether chest spotting is a signal in territorial interactions among male song sparrows. We experimentally increased or reduced the extent of spotting in males and measured the change in their aggression. We found that reduced-spotting males showed a more moderate seasonal decrease of aggression compared to males with enlarged spotting reduced aggression, possibly because the former experienced more intrusions later on in the breeding season while the latter experienced fewer intrusions. These results are consistent with chest spotting size in song sparrows functioning as a signal of territory holding potential of the bearer.

Infectious disease is linked to impaired cognition across a breadth of host taxa and cognitive abilities, potentially contributing to variation in cognitive performance within and among populations. Impaired cognitive performance can stem from direct damage by the parasite, the host immune response, or lost opportunities for learning. Moreover, cognitive impairment could be compounded by factors that simultaneously increase infection risk and impair cognition directly, such as stress and malnutrition. As highlighted in this review, however, answers to fundamental questions remain unresolved, including the frequency, duration, and fitness consequences of infection-linked cognitive impairment in wild animal populations, the cognitive abilities most likely to be affected, and the potential for adaptive evolution of cognition in response to accelerating emergence of infectious disease.

The costs and benefits of breeding behaviors are influenced by environmental conditions, and habitat variation can shift the degree to which behaviors are expressed. Novel urban habitats have been shown to differ significantly in disturbances such as noise, light at night, and human presence, as well as resource availability, compared to rural habitats. Perhaps because of these environmental differences, urban males of several species are consistently more aggressive than ruralmales, raising the hypothesis that greater territorial aggression is beneficial in urban habitats. Though often ignored, female songbirds of many species also perform aggressive territorial behaviors toward conspecifics during the breeding season. For sociallymonogamous songbirds, this aggression functions to ensure partner fidelity and secure resources for reproduction. Studies of the effects of urbanization on songbird behavior have yet to determine if urban females also express greater territorial aggression. Importantly, energetically demanding behaviors such as territoriality and parental care should constrain one another, leading to behavioral trade-offs during the breeding season. Though territorial aggression and parental care are inversely related in males of several species of songbird, this relationship is understudied in female songbirds, particularly those facing environmental change, such as urbanization. In this study, we compared aggressive signaling and a measure of parental care (maternal nest visitation rates) between female song sparrows (Melospiza melodia), living in urban and rural habitats. We hypothesized that female aggressive signaling would be higher in urban environments compared to rural, and negatively correlated with maternal visitation rates. We found that urban females, like males, expressed increased aggressive signaling compared to rural. However, female aggressive signaling was not related to our measure of maternal care, suggesting females aren't facing a trade-off between these two behaviors. Collectively, our results are consistent with the hypothesis that urban habitats promote territorial aggression in female song sparrows. As urbanization continues to spread, understanding the behavioral changes animals employ in urban environments requires studying individuals of different sexes and age classes, and will help us understand how some species are able to cope with human-induced rapid environmental change.

A central theme in the field of ecology is understanding how environmental variables influence a species’ distribution. In the last 20 years, there has been particular attention given to understanding adaptive physiological traits that allow some species to persist in urban environments. However, there is no clear consensus on how urbanization influences physiology, and it is unclear whether physiological differences in urban birds are directly linked to adverse outcomes or are representative of urban birds adaptively responding to novel environmental variables. Moreover, though low-density suburban development is the fastest advancing form of urbanization, most studies have focused on animals inhabiting high intensity urban habitats. In this study, we measured a suite of physiological variables that reflect condition and immune function in male song sparrows ( Melospiza melodia ) from rural and suburban habitats. Specifically, we measured hematological indices [packed cell volume (PCV), hemoglobin concentration, mean corpuscular hemoglobin concentration (MCHC)], circulating glutathione (total, reduced, and oxidized), oxidative damage (d-ROM concentration), antioxidant capacity, and components of the innate immune system [bacteria killing ability (BKA), white blood cell counts]. We also measured whole-animal indices of health, including body condition (scaled mass index length) and furcular fat. Song sparrows inhabiting suburban environments exhibited lower hemoglobin and MCHC, but higher body condition and furcular fat scores. Additionally, suburban birds had higher heterophil counts and lower lymphocyte counts, but there were no differences in heterophil:lymphocyte ratio or BKA between suburban and rural birds. PCV, glutathione concentrations, and oxidative damage did not differ between suburban and rural sparrows. Overall, suburban birds did not exhibit physiological responses suggestive of adverse outcomes. Rather, there is some evidence that sparrows from rural and suburban habitats exhibit phenotypic differences in energy storage and metabolic demand, which may be related to behavioral differences previously observed in sparrows from these populations. Furthermore, this study highlights the need for measuring multiple markers of physiology across different types of urban development to accurately assess the effects of urbanization on wildlife.

Urban habitats present animals with persistent disturbances and acute stressors not present in rural habitats or present at significantly lower levels. Differences in the glucocorticoid stress response could underlie colonization of these novel habitats. Despite urban habitats characterization as more stressful, previous comparisons of urban and rural birds have failed to find consistent differences in baseline and stress induced glucocorticoid levels. Another aspect of glucocorticoid regulation that could underlie an animal's ability to inhabit novel habitats, but has yet to be well examined, is more efficient termination of the glucocorticoid stress response which would allow birds in urban habitats to recover more quickly after a disturbance. The glucocorticoid stress response is terminated by negative feedback achieved primarily through their binding of receptors in the hippocampus and hypothalamus and subsequent decreased synthesis and release from the adrenals. We investigated if male song sparrows (Melospiza melodia) in urban habitats show more efficient termination of the glucocorticoid stress response than their rural counterparts using two approaches. First, we measured glucocorticoid receptor, mineralocorticoid receptor and 11β-HSD2 (an enzyme that inactivates corticosterone) mRNA expression in negative feedback targets of the brain (the hippocampus and hypothalamus) as a proxy measure of sensitivity to negative feedback. Second, we measured plasma corticosterone levels after standardized restraint and again following a challenge with the synthetic glucocorticoid, dexamethasone, as a means of assessing how quickly birds decreased glucocorticoid synthesis and release. Though there were no differences in the hypothalamus of urban and rural song sparrows, urban birds had lower glucocorticoid receptor and 11β-HSD2 mRNA expression in the hippocampus. Further, urban and rural birds had similar reductions in corticosterone following the dexamethasone challenge, suggesting that they do not differ in how quickly they decrease glucocorticoid synthesis and release. Thus, urban and rural song sparrows display similar termination of the glucocorticoid stress response even though urban birds have decreased hippocampal glucocorticoid receptor and 11β-HSD2 abundance.

Lead (Pb) is a pervasive global contaminant that interferes with sensitive windows for neurological development and causes oxidative damage to tissues. The effects of moderate and high exposure to Pb have been well-studied in birds, but whether low-level early-life exposure to Pb influences adult phenotype remains unclear. Female songbirds use a male's song and coloration to discriminate between high- and low-quality males. Therefore, if early-life exposure to Pb disrupts song learning ability or shifts the allocation of antioxidant pigments away from colorful secondary sexual traits, male birds exposed to Pb may be less attractive to females. We exposed developing zebra finches (Taeniopygia guttata) to Pb-contaminated drinking water (100 or 1000 parts per billion [ppb]) after hatching (days 0–100). Once male finches reached adulthood (120–150 days post hatch), we measured song learning ability, coloration of bill and cheek patches, and volume of song nuclei in the brain. We also measured female preference for Pb-exposed males relative to control males. Finally, we measured motoric and spatial cognitive performance in male and female finches to assess whether cognitive traits differed in their sensitivity to Pb exposure. Male zebra finches exposed to 1000 ppb Pb had impaired song learning ability, reduced volume of song nuclei, bills with less redness and received less attention from females. Additionally, Pb exposure impaired motoric performance in both male and female finches but did not affect performance in a spatial cognitive task. Adult finches exposed to Pb-contaminated water had higher blood-Pb levels, though in all cases blood-Pb levels were below 7.0 µg dL−1. This study suggests that low-level exposure to Pb contributes to cognitive deficits that persist into adulthood and may indirectly influence fitness by altering secondary sexual traits and reducing male attractiveness.