Veterinary ectoparasiticides widely used on companion animals often contain synthetic insecticides whose agricultural applications have been restricted in the European Union because of environmental concerns. These neurotoxic compounds can persist on animal fur and enter surrounding environments, potentially exposing non-target organisms. The Great Tit (Parus major), a common passerine in both urban and forest habitats, frequently incorporates animal fur in nest linings, thereby creating a potential exposure route for breeding adults and nestlings. We analysed fur-containing nest material collected from 63 Great Tit nests in artificial nest boxes at an urban site and a nearby protected forest in Hungary during the 2025 breeding season, sampling at mid-nestling stage and also after fledging. Using HPLC-MS/MS and GC-MS, we detected several veterinary ectoparasiticides in nest materials, including fipronil, fipronil sulfone, imidacloprid, and permethrin. Acetamiprid was found only in urban nests, indicating additional, non-veterinary environmental sources. Multiple insecticides were present in nest material, with higher contamination levels and a greater number of detected compounds in urban compared to forest nests. Residues were present at both sampling times but declined over the course of the breeding cycle. Although contamination was not associated with the measured reproductive parameters of Great Tits, our findings show that veterinary ectoparasiticides can contaminate wild bird nests, including those in protected forest ecosystems. This highlights a previously under-recognised pathway linking companion animal treatments to wildlife exposure and underscores the need to assess the ecological risks and trade-offs associated with widespread veterinary insecticide use.

Birth sex ratio biases can amplify extinction risks, especially in small, zoo-maintained populations which is of particular concern in species under threat of extinction. Thus, understanding the drivers of such biases is critical for conservation outcomes. We analysed birth records from 129 avian and 324 mammalian species in zoos worldwide between 1980 and 2021. Using Bayesian phylogenetic models, we found a phylogenetic signal in birth sex ratios (BSR), with substantial variation across clades. Penguins, falcons, and parrots showed slightly male-biased BSRs; ungulates showed female-biased BSR, and primates male-biased BSR. Across birds, variation in BSRs was predicted by sexual size dimorphism and clutch size, whereas in mammals, mating system was the main predictor of BSR. We identified 30 conservation flagship species with significantly biased BSRs, raising concern for the demographic sustainability of their captive populations. These results highlight the role of both evolutionary history and life-history traits in shaping sex ratio variation across taxa. Our findings underscore the importance of integrating phylogenetic and biological predictors into conservation planning and breeding program design. They also call for further research into the biological and management processes-that include sexual selection, parental investment, housing, and sexing practices-that may contribute to sex ratio variation in zoo populations.

This study provides rare evidence from a long-lived seabird in captivity, extending insights beyond the short-lived model species that dominate social research. By mapping aggressive interactions as social networks, we demonstrate that dominance rank in African penguins is not determined solely by age or sex: rearing conditions exert long-lasting effects on an individual’s position in the hierarchy. Contrary to the common assumption that artificial rearing impairs social competence, hand- and mix-reared birds consistently outperformed their parent-reared conspecifics. Our results also reveal the dynamic role of age, showing that younger penguins can dominate older individuals, reshaping how we understand the formation of social hierarchies. These findings have direct implications for conservation and zoo management, offering new strategies to improve welfare and social stability in captive penguin populations.

The increasing presence and activities of people in both urban environments and non‐urban areas result in the exposure of many wild animal populations to persistent human disturbance. As a response, individuals in disturbed populations often become tolerant towards humans, which can have significant ecological and societal consequences, for example by affecting ecosystem services and human–wildlife conflicts. Although several mechanisms have been proposed to explain the emergence and spread of disturbance tolerance, their biological bases are rarely investigated in natural populations. In this study, we investigated behavioural tolerance in great tits Parus major along urbanization gradients of two cities representing different levels of human disturbance. Specifically, we studied whether variation in disturbance tolerance is related to epigenetic variation in the DRD4 gene that is often linked to behavioural plasticity. We did not detect differences in DNA methylation at 23 CpG sites between wild great tit populations breeding in differently urbanized areas. However, variation in methylation at some CpG sites was associated with two proxies of tolerance, return latency and the vigilance behaviour of parent birds measured after standardized disturbance. These findings suggest that epigenetic variation may be involved in the processes generating behavioural tolerance to human disturbance. These results have implications for understanding the mechanisms by which animal populations can respond to disturbances in human‐dominated environments.

Sex chromosomes can determine male and female phenotypes, and the resulting sex differences may have significant impacts on ecology and life history. One manifestation of this link is that ZW/ZZ sex-determination systems are associated with more male-skewed adult sex ratio (ASR, proportion of males in the adult population) than XX/XY systems across tetrapods (amphibians, reptiles, birds, and mammals). Here, we investigate four demographic processes: male and female offspring production, sex differences in juvenile and adult mortalities and in timing of maturation that can contribute to ASR variation between XX/XY and ZW/ZZ systems, using phylogenetic analyses of a large dataset collected from tetrapod species in the wild. We show that sex differences in adult mortality reliably predict ASR that is also more male-biased in XX/XY species than in ZW/ZZ species. Sex differences in juvenile mortality and in maturation time also contribute to ASR skews, but do not differ consistently between XX/XY and ZW/ZZ systems. Phylogenetic path analyses confirm an influence of sex-determination system on ASR through sex-biased adult mortalities. Together these results infer that sex chromosomes can impact, via demographic pathways, frequency-dependent selection emerging from the relative number of males and females. We call for follow-up studies to uncover the potentially complex web of associations between sex determination, population dynamics, and social behavior.

Gene flow may be limited between urban and non-urban populations of wild animals that can influence their landscape-level genetic structure and potential to adapt to new ecological conditions. To test this idea, we genetically characterized great tit ( Parus major ) populations breeding in an urban and a forest area 3.5 km apart, differing in several phenotypic traits some of which may contribute to adaptation to urban living. We used 16 microsatellite markers to genotype 189 breeding adult individuals (119 urban and 70 forest birds) and (1) tested whether the two populations are genetically differentiated, and (2) estimated the rate and direction of migration between the sites. Heterozygosity tended to be lower in the urban than in the forest habitat. Genetic population structure analyses did not show a consistent clustering of breeding birds between the urban and forest sites, and this conclusion was not affected by the inclusion of phenotypic data in the analyses. The pairwise fixation index (F st ) was low (0.009) and only 1% of the total genetic variance was explained by variation between populations. Finally, there was detectable gene flow between the two areas, and its estimated values did not suggest asymmetry in the direction of migration. We conclude that great tits living in the city are genetically connected to the nearby forest population by reciprocal migration, which may explain the low level of genetic differentiation.

Habitat changes associated with urbanization have major and complex effects on wildlife. In birds, urban populations often have lower reproductive success but are able to maintain similar or higher densities than non-urban populations. One explanation proposed for this paradox is that higher survival of birds in cities may compensate for lower reproduction. We use a 9-year dataset and Cormack-Jolly-Seber models to compare annual variation in apparent survival probabilities of adult great tits (Parus major) at two forests and two urban sites located in Hungary. Our analyses tested the effects of sex, age, year, population density on apparent survival, after correcting for the probability of detection. Apparent survival of great tits varied between 0.122 and 0.736, with study site and year having the greatest influence. Unexpectedly, urbanization did not have a consistent effect: the sites with the lowest and highest estimates of survival were both urban habitats. Survival probabilities at the two forest sites were similar to each other but were ~0.15 lower than survival in the best urban site and ~0.1. higher than survival in the worst urban site. Survival probabilities exhibited marked inter-annual variation in all sites, although temporal patterns were not consistent among sites suggesting the variation was not driven by inter-annual variation in regional scale factors. Survival probabilities decreased with bird age at both urban sites in most years, but such patterns were not detected at forest sites. Our results demonstrate that the impacts of aging on avian survival rates can diverge between urban and forest habitats, and that the demographic factors regulating urban populations can vary between locations. Age-specific variation should be taken into account in urban ecology and further exploration of the factors driving the heterogeneity will help inform conservation of biodiversity along rural-urban gradients.

Males and females often have different roles in reproduction, although the origin of these differences has remained controversial. Explaining the enigmatic reversed sex roles where males sacrifice their mating potential and provide full parental care is a particularly long-standing challenge in evolutionary biology. While most studies focused on ecological factors as the drivers of sex roles, recent research highlights the significance of social factors such as the adult sex ratio. To disentangle these propositions, here, we investigate the additive and interactive effects of several ecological and social factors on sex role variation using shorebirds (sandpipers, plovers, and allies) as model organisms that provide the full spectrum of sex role variation including some of the best-known examples of sex-role reversal. Our results consistently show that social factors play a prominent role in driving sex roles. Importantly, we show that reversed sex roles are associated with both male-skewed adult sex ratios and high breeding densities. Furthermore, phylogenetic path analyses provide general support for sex ratios driving sex role variations rather than being a consequence of sex roles. Together, these important results open future research directions by showing that different mating opportunities of males and females play a major role in generating the evolutionary diversity of sex roles, mating system, and parental care.

Sex chromosomes determine male and female phenotypes, and the resulting sex differences can have significant impacts on ecology and life history. One manifestation of this link is that ZZ/ZW sex-determination systems are associated with more male-skewed adult sex ratio (ASR, proportion of males in the adult population) than XY/XX systems across tetrapods (amphibians, reptiles, birds, and mammals). Here we investigate four demographic processes: male and female offspring production, sex differences in juvenile and adult mortalities and in timing of maturation that can contribute to ASR variation between XY/XX and ZZ/ZW systems, using phylogenetic analyses of a large dataset collected from tetrapod species in the wild. We show that sex differences in adult mortality reliably predict ASR, and it is also more male-biased in XY/XX species than in ZZ/ZW species. Sex differences in juvenile mortality or in maturation time also contribute to ASR skews, but do not differ consistently between XY/XX and ZZ/ZW systems. Phylogenetic path analyses confirm an influence of sex-determination system on ASR through sex-biased adult mortality. Thus, these results infer that sex chromosomes can impact, via demographic pathways, frequency-dependent selection emerging from the relative number of males and females. We call for follow-up studies to uncover the potentially complex web of associations between sex determination, population dynamics, and social behaviour. ### Competing Interest Statement The authors have declared no competing interest.

Sex-biased mortality can occur in birds during development, for example due to sexual differences in energy requirement and/or environmental sensitivity, or the effects of sex hormones or sex differences in the expression of mutations linked to sex chromosomes. The extent of sex-bias in mortality may also be related to environmental conditions that influence offspring development and survival. Urban areas often provide poorer conditions for nestling development resulting in higher offspring mortality compared to natural areas, which may accelerate sex differences in offspring mortality in cities. To test this hypothesis, we examined the sex ratio of dead offspring in Great Tits (Parus major), using 427 samples of unhatched eggs and dead nestlings collected in two urban and two forest sites between 2013 and 2019. The ratio of males in the whole sample of dead offspring (56.9%) was significantly higher than expected by an 1:1 ratio, and the strongest sex biases were detected in urban areas (57.6% males) and in young nestlings (<14 days old, 59.0% males). However, the sex ratios of dead offspring did not differ significantly among study sites and between offspring developmental stages. 29.3% of unhatched eggs contained a visible embryo, and the proportion of embryo-containing unhatched eggs did not differ significantly between urban and forest study sites. These results suggest male-biased offspring mortality in Great Tits, and highlight the need of large datasets to detect subtle differences between habitats and developmental stages.

The adult sex ratio (ASR, the proportion of males in the adult population) is an emerging predictor of reproductive behaviour, and recent studies in birds and humans suggest it is a major driver of social mating systems and parental care. ASR may also influence genetic mating systems. For instance male-skewed ASRs are expected to increase the frequency of multiple paternity (defined here as a clutch or litter sired by two or more males) due to higher rates of coercive copulations by males, and/or due to females exploiting the opportunity of copulation with multiple males to increase genetic diversity of their offspring. Here, we evaluate this hypothesis in reptiles that often exhibit high frequency of multiple paternity although its ecological and life-history predictors have remained controversial. Using a comprehensive dataset of 81 species representing all four non-avian reptile orders, we show that increased frequency of multiple paternity is predicted by more male-skewed ASR, and this relationship is robust to simultaneous effects of several life-history predictors. Additionally, we show that the frequency of multiple paternity varies with the sex determination system: species with female heterogamety (ZZ/ZW sex chromosomes) exhibit higher levels of multiple paternity than species with male heterogamety (XY/XX) or temperature-dependent sex determination. Thus, our across-species comparative study provides the first evidence that genetic mating system depends on ASR in reptiles. We call for further investigations to uncover the complex evolutionary associations between mating systems, sex determination systems and ASR.

Sex-biased mortality can occur in birds during development, for example due to sexual differences in energy requirement and/or environmental sensitivity, or the effects of sex hormones or sex differences in expression of mutations linked to sex chromosomes. Urban habitats often provide poorer conditions for nestling development resulting in higher offspring mortality compared to natural habitats, which may accelerate sex differences in offspring mortality in cities. To test this hypothesis, we examined sex-specific offspring mortality in great tits (Parus major), using 660 samples of dead offspring collected in two urban and two forest sites between 2013-2019. Overall, the sex ratio of dead offspring was significantly male-biased (56.80%). When habitats and age groups were analysed separately, the sex ratio of dead offspring was significantly male-biased in urban habitat (57.60%) and in young nestlings (58.62%), and non-significantly in the forest habitat (56.58%) and in unhatched embryos (54.11%) or in old nestlings (54.55%). However, these estimates were associated with wide confidence intervals, thus sex ratios of dead offspring did not differ significantly among study sites and between age groups when these were analysed together. 70.30% of unhatched eggs was not fertilized, and their proportion did not differ between urban and forest habitats. These results suggest male-biased offspring mortality in great tits, and highlight the need of large datasets to detect subtle differences between habitats and developmental stages.

Climate change and urbanisation are among the most salient human-induced changes affecting Earth’s biota. Extreme weather events can have high biological impacts and are becoming more frequent recently. In cities, the urban heat island can amplify the intensity and frequency of hot weather events. However, the joint effects of heat events and urban microclimate on wildlife are unclear, as urban populations may either suffer more from increased heat stress or may adapt to tolerate warmer temperatures. Here, we test whether the effects of hot weather on reproductive success of great tits (Parus major) are exacerbated or dampened in urban environments compared to forest habitats. By studying 760 broods from two urban and two forest populations over 6 years, we show that 14–16 days-old nestlings have smaller body mass and tarsus length, and suffer increased mortality when they experience a higher number of hot days during the nestling period. The negative effects of hot weather on body mass and survival are significantly stronger in forests than in urban areas, where these effects are dampened or even reversed. These results suggest that urban nestlings are less vulnerable to extreme hot weather conditions than their non-urban conspecifics. This difference might be the result of adaptations that facilitate heat dissipation, including smaller body size, altered plumage and reduced brood size. Alternatively or additionally, parental provisioning and food availability may be less affected by heat in urban areas. Our findings suggest that adaptation to heat stress may help birds cope with the joint challenges of climate change and urbanisation.

Urban animals often show bolder behaviour towards humans than their nonurban conspecifics. How-ever, it is unclear to what extent this difference is due to consistent individual characteristics or to plasticity such as habituation. To address this question, we investigated parental risk-taking behaviour in 371 female great tits in urban and forest populations by checking their nest repeatedly (several times per week, for up to nine breeding episodes) and recording their behavioural responses to this recurring disturbance during incubation. We found that urban females were bolder, as they stayed on the nest more often than females in forests. Furthermore, great tits produced alarm calls around the nests more frequently in urban than in forest habitats. There was no habitat difference in the frequency of an antipredator behaviour, the hissing threat display on the nest, although this was rare in both habitats. We also tested the consistency and plasticity of risk-taking behaviour on three different temporal scales (within breeding attempts, between broods within a year and across years). Staying on the nest was highly repeatable within females, whereas alarm calls had low repeatability within pairs at all three temporal scales. The probability of staying on the nest increased within breeding attempts, whereas the probability of alarm calls increased across years. Neither consistency nor plasticity in these components of risk taking differed between urban and forest habitats. We conclude that urban birds are bolder in multiple behavioural measures and, overall, both stable individual differences and behavioural plasticity may have contributed to the higher risk taking we often see in urban populations. Furthermore, staying on the nest appears to be an individually consistent trait in female great tits regardless of habitat ur-banization, providing a low-impact measurement of risk taking, which may potentially facilitate field studies related to individual differences in behaviour. (c) 2021 The Authors. Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour. This is an open access article under the CC BY license (http://creativecommons.org/licenses/ by/4.0/).

Body size often differs between the sexes (leading to sexual size dimorphism, SSD), as a consequence of differential responses by males and females to selection pressures. Adult sex ratio (ASR, the proportion of males in the adult population) should influence SSD because ASR relates to both the number of competitors and available mates, which shape the intensity of mating competition and thereby promotes SSD evolution. However, whether ASR correlates with SSD variation among species has not been yet tested across a broad range of taxa. Using phylogenetic comparative analyses of 462 amniotes (i.e., reptiles, birds, and mammals), we fill this knowledge gap by showing that male bias in SSD increases with increasingly female-skewed ASRs in both mammals and birds. This relationship is not explained by the higher mortality of the larger sex because SSD is not associated with sex differences in either juvenile or adult mortality. Phylogenetic path analysis indicates that higher mortality in one sex leads to skewed ASR, which in turn may generate selection for SSD biased toward the rare sex. Taken together, our findings provide evidence that skewed ASRs in amniote populations can result in the rarer sex evolving large size to capitalize on enhanced mating opportunities.

The ubiquitous activity of humans is a fundamental feature of urban environments affecting local wildlife in several ways. Testing the influence of human disturbance would ideally need experimental approach, however, in cities, this is challenging at relevant spatial and temporal scales. Thus, to better understand the ecological effects of human activity, we exploited the opportunity that the city-wide lockdowns due to the COVID-19 pandemic provided during the spring of 2020. We assessed changes in reproductive success of great tits (Parus major) at two urban habitats affected strikingly differently by the ‘anthropause’, and at an unaffected forest site. Our results do not support that urban great tits benefited from reduced human mobility during the lockdown. First, at one of our urban sites, the strongly (− 44%) reduced human disturbance in 2020 (compared to a long-term reference period) did not increase birds’ reproductive output relative to the forest habitat where human disturbance was low in all years. Second, in the other urban habitat, recreational human activity considerably increased (+ 40%) during the lockdown and this was associated with strongly reduced nestling body size compared to the pre-COVID reference year. Analyses of other environmental factors (meteorological conditions, lockdown-induced changes in air pollution) suggest that these are not likely to explain our results. Our study supports that intensified human disturbance can have adverse fitness consequences in urban populations. It also highlights that a few months of ‘anthropause’ is not enough to counterweight the detrimental impacts of urbanization on local wildlife populations.

Abstract The ubiquitous activity of humans is a fundamental feature of urban environments affecting local wildlife in several ways. Testing the influence of human disturbance would ideally need experimental approach, however, in cities, this is challenging at relevant spatial and temporal scales. Thus, to better understand the ecological effects of human activity, we exploited the opportunity that the city-wide lockdowns due to the COVID-19 pandemic provided during the spring of 2020. We assessed changes in reproductive success of great tits (Parus major) at two urban habitats affected strikingly differently by the ‘anthropause’ and at an unaffected forest site. Although anecdotic observations suggested that urban wildlife may benefit from reduced human mobility during the lockdown, our results do not support this. First, at one of our urban sites, the strongly (-44%) reduced human disturbance in 2020 (compared to a long-term reference period) did not increase birds’ reproductive output relative to the forest habitat where human disturbance was low in all years. Second, in the other urban habitat, recreational human activity considerably increased (+ 40%) during the lockdown and this was associated with strongly reduced nestling body size compared to the pre-COVID reference year. Analyses on meteorological conditions and the lockdown-induced changes in air pollution suggest that these factors are not likely to explain our results. Our study supports that intensified human disturbance can have adverse fitness consequences in urban populations. It also highlights that a few months of ‘anthropause’ is not enough to counterweight the detrimental impacts of urbanization on local wildlife populations.

Urban areas differ from natural habitats in several environmental features that influence the characteristics of animals living there. For example, birds often start breeding seasonally earlier and fledge fewer offspring per brood in cities than in natural habitats. However, longer breeding seasons in cities may increase the frequency of double-brooding in urban compared with nonurban populations, thus potentially increasing urban birds' annual reproductive output and resulting in lower habitat difference in reproductive success than estimated by studies focusing on first clutches only. In this study, we investigated 2 urban and 2 forests great tit Parus major populations from 2013 to 2019. We compared the probability of double-brooding and the total number of annually fledged chicks per female between urban and forest habitats, while controlling for the effects of potentially confounding variables. There was a trend for a higher probability of double-brooding in urban (44% of females) than in forest populations (36%), although this was not consistent between the 2 urban sites. Females produced significantly fewer fledglings annually in the cities than in the forest sites, and this difference was present both within single- and double-brooded females. Furthermore, double-brooded urban females produced a similar number of fledglings per season as single-brooded forest females. These results indicate that double-brooding increases the reproductive success of female great tits in both habitats, but urban females cannot effectively compensate in this way for their lower reproductive output per brood. However, other mechanisms like increased post-fledging survival can mitigate habitat differences in reproductive success.

Rapidly increasing urbanisation is one of the most significant anthropogenic environmental changes which can affect demographic traits of animal populations, for example resulting in reduced reproductive success. The food limitation hypothesis suggests that the shortage of high-quality nestling food in cities is a major factor responsible for the reduced reproductive performance in insectivorous birds. To study this explanation, we collected data on the parental provisioning behaviour of urban and forest great tits (Parus major) in three years that varied both in caterpillar availability (the main food of great tit nestlings) and in reproductive success of the birds. In all years, urban parents provisioned caterpillars in a smaller proportion to their nestlings, but the total amount of food per nestling (estimated by the volumes of all prey items) did not differ between habitats. In the two years with much lower reproductive success in urban than forest habitats, urban parents had higher provisioning rates, but provided more non-arthropod food and brought smaller prey items than forest parents. In the year with reduced habitat difference in reproductive success, urban parents were able to compensate for the scarcity of caterpillars by provisioning other arthropods rather than non-arthropod food, and by delivering larger preys than in the other years. Specifically, in this latter year, caterpillars provisioned by urban pairs were cc. twice as large as in the other two years, and were similar in size to caterpillars provisioned in the forest broods. These results show that although urban great tit parents can provide the same quantity of food per nestling as forest parents by reducing their brood size and increasing the per capita feeding rates for nestlings, they cannot compensate fully for the scarcity of high-quality preys (caterpillars) in poor years. In some years, however, favourable conditions for urban caterpillar development can greatly reduce food limitation in cities, allowing urban birds to achieve higher reproductive success. We suggest that urban green areas designed and managed in a way to facilitate conditions for phytophagous arthropods could improve habitat quality for urban birds.

Ern} o Vincze a, b, , Veronika B okony , L aszl o Zsolt Garamszegi d, , G abor Seress , Ivett Pipoly a, , Csenge Sinkovics , Krisztina S andor , Andr as Liker a, b a MTA-PE Evolutionary Ecology Research Group, University of Pannonia, Veszpr em, Hungary b Behavioural Ecology Research Group, Center for Natural Sciences, University of Pannonia, Veszpr em, Hungary c Lendület Evolutionary Ecology Research Group, Plant Protection Institute, Centre for Agricultural Research, E€ otv€ os Lor and Research Network, Budapest, Hungary d Institute of Ecology and Botany, ELKH Centre for Ecological Research, V acr at ot, Hungary e MTA-ELTE, Theoretical Biology and Evolutionary Ecology Research Group, Department of Plant Systematics, Ecology and Theoretical Biology, E€ otv€ os Lor and University, Budapest, Hungary