Search

**Abstract** Acoustic and visual signals are well known to play important roles in social communication in birds. Growing evidence suggests that many bird species, including species of songbirds, additionally have a well-developed sense of smell. However, we are still at the beginning of understanding the potential importance of chemical communication in the social lives of birds, for example in mate choice. The secretion of the preen gland may be an important contributor to the chemical phenotype of birds. Here, we report on a first characterisation of the chemical composition of the preen gland secretion of the Blue Tit (Cyanistes caeruleus), a common songbird which is an often used model species in animal behaviour and ecology, in particular also in studies of sexual selection and (extra-pair) mate choice. We found sex differences in the composition of the preen gland secretion in breeding Blue Tits. Females further tended to have a larger number of putative compounds in their secretions compared to males. We briefly discuss the possible implications of these findings and speculate that the chemical composition of the preen gland secretion may be a sexually selected trait in Blue Tits. Our preliminary findings warrant follow-up research into the patterns of within- and among individual variation in the chemical composition of the preen gland secretion as well as the identification of the main chemical compounds involved., **Zusammenfassung**Chemische Analyse zeigt Geschlechtsunterschiede im Bürzeldrüsensekret brütender BlaumeisenSingvögel sind vor allem für ihren Gesang und ihre Gefiedermerkmale, die bei der sozialen Kommunikation eine bedeutende Rolle spielen, bekannt. Immer mehr Hinweise deuten aber darauf hin, dass viele Vogelarten, darunter auch Singvogelarten, zusätzlich einen gut entwickelten Geruchssinn haben. Welche Bedeutung die geruchliche Kommunikation im sozialen Leben der Vögel, zum Beispiel bei der Partnerwahl, spielt ist allerdings weitgehend noch unbekannt. Das Bürzeldrüsensekret könnte in diesem Zusammenhang einen wichtigen Beitrag zum chemischen Phänotyp der Vögel leisten. In unserer Studie haben wir uns die chemische Zusammensetzung des Bürzeldrüsensekrets der Blaumeise (Cyanistes caeruleus) in der Brutzeit angeschaut. Blaumeisen sind eine viel genutzte Modellart in der Verhaltensforschung und Verhaltensökologie, insbesondere auch in Studien zur sexuellen Selektion und (außerpaarigen) Partnerwahl. Anhand von chemischen Analysen mittels Gas-Chromatographie fanden wir heraus, dass es bei brütenden Blaumeisen einen Geschlechtsunterschied in der Zusammensetzung des Bürzeldrüsensekrets gibt. Weibchen neigen außerdem dazu, eine größere Anzahl von vermeintlichen Substanzen in ihren Sekreten zu haben als Männchen. Wir diskutieren hier kurz die möglichen Implikationen dieser Ergebnisse und spekulieren, dass die chemische Zusammensetzung des Bürzeldrüsensekrets ein sexuell selektiertes Merkmal bei Blaumeisen sein könnte. Unsere vorläufigen Ergebnisse rechtfertigen Folgeuntersuchungen, in denen die Identifikation der wichtigsten chemischen Verbindungen des Bürzeldrüsensekrets und die Variation innerhalb und zwischen Individuen in der chemischen Zusammensetzung des Drüsensekrets im Fokus stehen.

Geographic variation of sexually selected male traits is common in animals. Female choice also varies geographically and several studies found female preference for local males, which is assumed to lead to local adaptation and, therefore, increases fitness. As females are the nondispersing sex in most mammalian taxa, this preference for local males might be explained by the learning of male characteristics. Studies on the preference of females in female-dispersing species are lacking so far. To find out whether such females would also show preferences for local males, we conducted a study on greater sac-winged bats (Saccopteryx bilineata), a species where females disperse and males stay in their natal colony. Male greater sac-winged bats possess a wing pouch that is filled with odoriferous secretion and fanned toward females during courtship display. In a combination of chemical analysis and behavioral preference tests, we analyzed whether the composition of wing sac secretion varies between two geographically distinct populations (300 km), and whether females show a preference for local or distant male scent. Using gas chromatography, we found significant differences in the composition of the wing sac odors between the two geographically distinct populations. In addition, the behavioral preference experiments revealed that females of both populations preferred the scent of geographically distant males over local males. The wing sac odor might thus be used to guarantee optimal outbreeding when dispersing to a new colony. This is—to our knowledge—the first study on odor preference of females of a species with female-biased dispersal.

Individual life histories are strongly influenced by early environmental conditions and experiences. They shape morphology as well as behaviour and can promote adaptive divergence and phenotypic plasticity with regard to different habitat types. The fire salamander (Salamandra salamandra) in the Kottenforst forest in Bonn, Germany, exhibits two genetically distinct ecotypes occurring in two larval habitats, either ponds or streams. In this study, we investigated whether both ecotypes differ in risk-taking behaviour, measured as the behavioural response during a shelter-emergence test and a shelter-seeking test, and whether larval habitat type and size impact these behaviours. Our results revealed an influence of size as well as habitat type. Larger larvae of both habitats appear to be more risk-prone, as they spent more time outside the starting shelter in the shelter-emergence test. Irrespective of size, pond larvae sought shelter more often in the shelter-seeking test and are thus considered to be less risk-prone. These results indicate that larvae conform to a given niche by adjusting their behaviour. Future studies are needed to disentangle the role of genetic adaptation or phenotypic plasticity and to investigate long-term consequences of the larval habitat for the adult phenotype. Thereby, efforts should be made to create a concise set of multiple tests assessing behavioural patterns.

Experiments from our research group have demonstrated that the olfactory sense of birds, which has been considered as unimportant for a long time, plays a prominent role as communication channel in social behaviour. Odour cues are used e.g. by zebra finch chicks to recognize the mother, by adult birds to distinguish their own eggs from others, or to recognize kin. While there is quite a lot of evidence for the importance of odour for social behaviour, it is not known as yet which brain areas may be involved in the processing of socially relevant odours. We therefore compared the brain activation pattern of zebra finch males exposed to their own offspring odour with that induced by a neutral odour stimulus. By measuring head saccade changes as behavioural reaction and using the expression of the immediate early gene product c-Fos as brain activity marker, we show here that the activation pattern, namely the activity difference between the left and the right hemisphere, of several hippocampal areas in zebra finch males is altered by the presentation of the odour of their own nestlings. In contrast, the nucleus taeniae of the amygdala (TnA) exhibits a tendency of a reduction of c-Fos activation in both hemispheres as a consequence of exposure to the nestling odour. We conclude that the hippocampus is involved in odour based processing of social information, while the role of TnA remains unclear.

Urbanisation is a major anthropogenic perturbation that progressively alters multiple environmental parameters, thereby presenting novel evolutionary challenges to wild populations. Symbiotic microorganisms residing in the gastrointestinal tracts (gut) of vertebrates have well-established mutual connections with the physiology of their hosts and respond quickly to environmental alterations. However, the impacts of anthropogenic changes and urbanisation on gut microbiota remain poorly understood, especially in early development. To address this knowledge gap, we investigated the gut microbiota in juvenile great tits (Parus major) in an urban mosaic. First, we compared the microbiota of nestlings reared in artificial nest boxes or natural cavities. Next, we analysed microbiota variations employing two distinct urbanisation frameworks, (i) the classical urban/rural dichotomy and (ii) gradual changes in the amount of impervious surface area (ISA) in the urban space and identified the environmental variables that predicted the changes in the microbial communities. Alpha diversity was influenced by cavity type and by the amount of impervious surface surrounding the breeding location. Alpha diversity was also positively correlated with the tree cover density. Beta diversity differed between urban and rural sites, and these alterations covaried with sound pollution, the distance to the city centre and the distance to the closest road. Overall, the microbial communities reflect and are possibly influenced by the heterogeneous environmental modifications that are typical of the urban space. Strikingly, the choice of framework used to define the urban space can influence the outcomes of studies investigating animal-microbe symbiosis. Our results open new perspectives from which the potential function of microbial symbionts in the adaptation of their hosts to anthropogenic stress can be investigated.

Mate attraction via chemosignalling is common in group-living mammals. In the greater short-nosed fruit bats, Cynopterus sphinx, it remains unclear whether males use chemosignals to attract females. Here, we use field observations to describe patterns of male chemosignalling in this species, then use preference test in captivity to infer the function of chemosignals. We observed that males engage in chemosignalling through wing fanning after marking their leaf ‘tent' roosts with saliva. The male fanned each wing alternatively and vigorously and performed frequent tongue flicks to the wing membrane. Such behaviour is exhibited towards females that returned to the tent. Afterwards, the male embraced the female with both wings. Wing fanning and tent marking occurred more often during the mating season than the non-mating season. To infer the role of chemosignals in recruiting females, we performed two different choice tests. In the first test, we tested whether females preferred the tent containing cotton balls that were rubbed gently and repeatedly on the ventral side of the plagiopatagium of the male, versus a control tent without any conspecific odour. In the second test, we tested whether females preferred a tent that was marked by a male's saliva versus a control tent without such markings. The result of both tests indicated that females recognized the saliva and ‘scent’ from the wings of males, since females always preferred tents with male chemosignals over the controls. Our findings suggest that the recruitment of females and subsequent formation of a harem in the greater short-nosed fruit bats is influenced by male chemosignals.

The outcomes of ecological interactions fall along a continuum from cooperative (mutually beneficial) to antagonistic (detrimental to one or both parties). This applies at both interspecific (e.g. plant-animal interactions) and intraspecific levels (e.g. male-female interactions). Phenotypic variation among individuals is increasingly being recognised as an important factor in ecological processes, and trait variation in either interacting party may determine the outcome of the interaction, such as whether a symbiont provides net costs or benefits to their host, or whether two conspecifics decide to cooperatively forage or to compete over food. This systematic review investigates the role of intraspecific trait variation (‘ITV’) in determining the cooperative-antagonistic outcome of ecological interactions. Based on a literature collection of 95 empirical and theoretical publications meeting our inclusion criteria, we give an overview of the various mechanisms that can lead to shifts between antagonism and cooperation within or between species. We describe two broad classes of interrelated mechanisms that may drive shifts in outcomes. First, trait frequency effects occur when processes influencing a population’s composition of traits linked to cooperation or antagonism (e.g. aggressive personality types, cheater/exploiter phenotypes etc.) lead to net shifts in interaction outcome. Second, systemic variance effects occur when changes in the amount of ITV in the population (as opposed to the mean phenotype) is the factor driving shifts. Both heritable genetic differences among individuals and phenotypic plasticity are important sources of phenotypic variation. The specific mix of heritable vs. plastic ITV may determine whether a change from cooperative to antagonistic, or vice versa, is likely to be short-term (i.e. context-dependent) or lead to more persistent shifts (e.g. mutualism breakdown). To guide future research on this topic we describe knowledge gaps and divergences between empirical and theoretical literature, further highlighting the value of applying research synthesis methods in ecology and evolution.

Predation is one of the strongest selection pressures, forcing prey organisms to detect predators and to display various antipredator behaviours, such as refuge-use or decreased activity. To recognise predators, chemosensory cues play a pivotal role, particularly in aquatic ecosystems. However, it is less known whether the ability to use these cues to respond with adequate antipredator behaviour varies between individuals occupying different habitats that are dissimilar in predation risk. Using field experiments, we examined antipredator behaviour of larval fire salamanders (Salamandra salamandra) from two different habitats, ponds and streams. Among other differences, ponds and streams are inhabited by habitat-specific predators, such as alpine newts (Ichthyosaura alpestris) occurring in ponds. We exposed larvae from both habitats to either chemical cues from alpine newts or a blank control (tap water) and investigated potential differences in their behavioural responses in two experiments. Pond larvae, but not stream larvae, became significantly less active when faced with chemical cues from newts compared to those faced with a control stimulus. Moreover, larvae from both habitats tested in water containing chemical cues spent significantly less time outside a shelter than those in control water. Our results demonstrate that larval fire salamanders recognise predatory newts through kairomones and alter their behaviour accordingly. However, experience with predatory newts may not be necessary to differentiate kairomones from control water, but may be beneficial for larvae to further develop their antipredator behaviour, thus representing conformance to a niche.

Replication studies are essential for assessing the validity of previous research findings and for probing their generality. However, it has proven challenging to reproduce the results of ecological and evolutionary studies, partly because of the complexity and lability of many of the phenomena being investigated, but also due to small sample sizes, low statistical power and publication bias. Additionally, replication is often considered too difficult in field settings where many factors are beyond the investigator’s control and where spatial and temporal dependencies may be strong. We investigated the feasibility of reproducing original research findings in the field of chemical ecology by attempting to replicate a previous study by our team on Antarctic fur seals (Arctocephalus gazella). In the original study, skin swabs from 41 mother-offspring pairs from two adjacent breeding colonies on Bird Island, South Georgia, were analysed using gas chromatography-mass spectrometry. Seals from the two colonies differed significantly in their chemical fingerprints, suggesting that colony membership may be chemically encoded, and mothers were also chemically similar to their pups, implying that phenotype matching may be involved in mother-offspring recognition. Here, we generated and analysed comparable chemical data from a non-overlapping sample of 50 mother-offspring pairs from the same two colonies five years later. The original results were corroborated in both hypothesis testing and estimation contexts, with p-values remaining highly significant and effect sizes, standardized between studies by bootstrapping the chemical data over individuals, being of comparable magnitude. We furthermore expanded the geographic coverage of our study to include pups from a total of six colonies around Bird Island. Significant chemical differences were observed in the majority of pairwise comparisons, indicating not only that patterns of colony membership persist over time, but also that chemical signatures are colony-specific in general. Our study systematically confirms and extends our previous findings, while also implying that temporal and spatial heterogeneity need not necessarily negate the reproduction and generalization of ecological research findings.

Warning colouration reduces predation risk by signalling or mimicking the unpleasantness of prey and therefore increases survival. We tested in two experiments the evolutionary costs and benefits of the yellow colour pattern in fire salamanders (Salamandra salamandra), which display a yellow/black colour pattern usually associated with toxic alkaloids. Our first experiment aimed to test whether the development of colouration is condition dependent and thus related to developmental costs, i.e. influenced by resource availability during the developmental process. Therefore, we reared fire salamander larvae under different nutritional conditions and compared the relative amount of yellow they developed after metamorphosis. Fire salamander larvae reared under limited food conditions had a lower proportion of yellow following metamorphosis than control larvae reared under superior food conditions. In a second experiment we tested whether the proportion of yellow has an impact on the risk of being attacked using artificial models. We tested, in salamander-free and salamander-occupied natural habitats, whether artificial clay models with different proportions of yellow and black receive different attack rates from potential predators (birds, mammals, insects). In clay models the proportion of yellow and the site had a significant effect on predation risk. Models with larger amounts of yellow had fewer bite marks from predators such as carabid beetles and birds, but only in sympatry with salamanders. In conclusion, the early expression of conspicuous colouration seems to be condition dependent and therefore potentially costly. Furthermore, the yellow colouration of fire salamanders act as a signal that potentially reduces their risk of being attacked by predators. Thus, the yellow colouration of fire salamanders seems to represent an adaptive trait that reduces the risk of predation, which can be expressed in higher quantity by individuals of a certain condition.

Infochemicals, including hormones, pheromones, and allelochemicals, play a central role in mediating information and shaping interactions within and between individuals. Due to their high plasticity, infochemicals are predestined mediators in facilitating individualized niches of organisms. Only recently it has become clear that individual differences are essential to understand how and why individuals realize a tiny subset of the species' niche. Moreover, individual differences have a central role in both ecological adjustment and evolutionary adaptation in a rapidly changing world. Here we highlight that infochemicals act as key signals or cues and empower the realization of the individualized niche through three proposed processes: niche choice, niche conformance, and niche construction.

Olfactory nest recognition has been demonstrated in several avian species, including some gregarious Estrildid finches. In social species, which also form breeding colonies, olfactory nest recognition gives parents and fledglings the opportunity to identify the natal nest. However, in less social birds, such as Diamond Firetails, which rarely breed in colonies, the benefit from olfactory nest recognition is potentially less pronounced. Using a set-up that has successfully been used in odour discrimination tests in other Estrildid finches, we tested whether Diamond Firetail parents and fledglings recognise the scent of their own nest. We found no evidence of olfactory nest recognition, either in parents or in fledged juveniles. This might indicate that Diamond Firetails have limited olfactory abilities, or that they utilise their olfactory capacities in other contexts which may be related to a certain extent to their lifestyle which is less social than that of other closely related species. Our results reveal new insights into potential selective pressures underlying the interspecific differences in olfactory nest recognition among Estrildid finches, and how the species-specific ecology may have shaped sensory recognition.

In birds, the preen gland produces a waxy secretion that protects the plumage from environmental conditions and ectoparasites and is, therefore, important for feather maintenance, individual quality, and health. Several studies have demonstrated that the size of the preen gland and the composition of its secretion change throughout the course of reproduction, thus indicating an increased requirement for preen gland secretion during the breeding season. However, the exact timing of the gland's size increase is unknown. To get information about this temporal pattern, we repeatedly measured the preen glands in 22 breeding pairs of adult domesticated Zebra Finches (Taeniopygia guttata) during the course of reproduction (prior to the breeding season, on the day the first egg was laid, on the day the first chick hatched, on the day the first chick fledged, and after the chicks became independent). Zebra Finches have biparental care, in which both sexes are involved in incubation and food provisioning. ...

Summary Offspring often solicit, and compete for, limited parental care by elaborate begging behaviour. Kin selection theory predicts that competing offspring should modify the intensity of their begging depending on the degree of relatedness to their nest- or litter-mates. Empirical evidence in birds, which are a key model in the study of parent-offspring interactions, indeed indicates that a lower level of relatedness between offspring in the nest correlates with more intense begging (i.e. more ‘selfish’ behaviour). This implies that competing nestlings can recognize kin, but the mechanism underlying such discrimination is unclear. Birds have long been thought to mainly rely on visual and auditory cues in their social communication, but there is now growing evidence for the importance of olfactory cues too. To assess the potential importance of olfactory cues in modulating nestling begging behaviour, we experimentally tested in a free-living bird, the blue tit Cyanistes caeruleus, if nestlings discriminate and adjust their begging behaviour depending on their familiarity with a conspecific nestling odour stimulus. We found that individuals responded with longer and more intense begging bouts to an unfamiliar compared with a familiar odour stimulus. Our findings provide first evidence for a role of olfaction in modulating offspring begging behaviour in a wild bird population. Although our experiment cannot differentiate between the effects of familiarity and relatedness, it raises the interesting possibility that blue tit nestlings may also discriminate between odours of close kin and less related individuals, and adjust their begging behaviour accordingly. This hypothesis requires further testing. A lay summary is available for this article.

Among mammals, bats exhibit extreme variation in sociality, with some species living largely solitary lives while others form colonies of more than a million individuals. Some tropical species form groups during the day that persist throughout the year while many temperate species only gather into groups during hibernation or parturition. How groups form and then persist has now been described for a number of species, but the degree to which kinship explains patterns of association has never been quantified across species. Here, we use social network analysis and genetic data to determine the extent to which relatedness contributes to associations among individuals estimated from free-ranging animals across nine species from four families of bats. Network analysis reveals that all species show evidence of emergent social structure. Variation in the strength of the relationship between genetic relatedness and social association appears to be related to the degree of roost switching, i.e., species in which individuals change roosts frequently tend to exhibit higher levels of association among relatives. Sex-biased dispersal determines whether associations were between male or female relatives. The strength of associations among kin does not predict known occurrence of complex behaviors, such as dominance or various types of cooperation, indicating that kinship is not a prerequisite for social complexity in bats.Significance statementThe number of differentiated relationships has been proposed as a way to measure social complexity. Among primates, relationships can be differentiated on the basis of rank, age, kinship, or association. Application of this approach to other groups of mammals that vary in sociality could help reveal ecological, behavioral, or cognitive similarities and differences between species. As a first step toward this approach, we used social network analysis on long-term individual records and estimated relatedness using genetic markers for nine species of bats. We confirmed nonrandom emergent social structure in all species. Kinship was predictive of social association among individuals of the same sex in a few species, but largely independent of the occurrence of complex behaviors, such as dominance among males or cooperation among females. Complex social behavior in bats appears to require frequent interactions among a small number of individuals that roost together for multiple years.

Early developmental conditions can have a strong influence on the life history. Fire salamanders represent an interesting system due to their biphasic life cycle consisting of an aquatic larval stage and a subsequent terrestrial stage. The environment experienced as larvae affects growth, age at metamorphosis and behaviour. In particular, diet restrictions can cause developmental stress. However, the impact of such developmental stress on the growth and behaviour in a long-lived amphibian are not yet well studied. We manipulated the early diet of sibling pairs of fire salamander larvae. One sibling was raised under food-restriction and the other was raised under conditions of abundant food. We report long-term effects on growth and exploratory behaviour; exploratory behaviour was tested in two sessions, first as young juveniles and later as sub-adults. We found that food-restricted conditions during early development affected exploratory behaviour. Early deficits in body mass and body size were compensated later in life, by approximately 18 months of age. When morphological differences were compensated, the initial differences in exploratory behaviour also disappeared. Thus, compensation not only led to an alignment of body parameters but was also accompanied by an adjustment in exploratory behaviour. No cost of compensation was detected, but future research will need to verify whether the potential costs of compensation are negligible in slow-growing salamanders or whether they are shifted to later life stages. Our study provides novel insights into the life history of fire salamanders and reveals that early larval conditions have effects on individuals long after metamorphosis.

Dystonia is a heterogeneous group of hyperkinetic movement disorders. The unifying descriptor of dystonia is the motor manifestation, characterized by continuous or intermittent contractions of muscles that cause abnormal movements and postures. Additionally, there are psychiatric, cognitive, and sensory alterations that are possible or putative non-motor manifestations of dystonia. The pathophysiology of dystonia is incompletely understood. A better understanding of dystonia pathophysiology is highly relevant in the amelioration of significant disability associated with motor and non-motor manifestations of dystonia. Recently, diminished olfaction was found to be a potential non-motor manifestation that may worsen the situation of subjects with dystonia. Yet, this finding may also shed light into dystonia pathophysiology and yield novel treatment options. This article aims to provide background information on dystonia and the current understanding of its pathophysiology, including the key structures involved, namely, the basal ganglia, cerebellum, and sensorimotor cortex. Additionally, involvement of these structures in the chemical senses are reviewed to provide an overview on how olfactory (and gustatory) deficits may occur in dystonia. Finally, we describe the present findings on altered chemical senses in dystonia and discuss directions of research on olfactory dysfunction as a marker in dystonia.

Despite an increasing appreciation of the importance of host-microbe interactions in ecological and evolutionary processes, the factors shaping microbial communities in wild populations remain poorly understood. We therefore exploited a natural experiment provided by two adjacent Antarctic fur seal (Arctocephalus gazella) colonies of high and low social density and combined 16S rRNA metabarcoding with microsatellite profiling of mother-offspring pairs to investigate environmental and genetic influences on skin microbial communities. Seal-associated bacterial communities differed profoundly between the two colonies, despite the host populations themselves being genetically undifferentiated. Consistent with the hypothesis that social stress depresses bacterial diversity, we found that microbial alpha diversity was significantly lower in the high-density colony. Seals from one of the colonies that contained a stream also carried a subset of freshwater-associated bacteria, indicative of an influence of the physical environment. Furthermore, mothers and their offspring shared similar microbial communities, in support of the notion that microbes may facilitate mother-offspring recognition. Finally, a significant negative association was found between bacterial diversity and heterozygosity, a measure of host genetic quality. Our study thus uncovers a complex interplay between environmental and host genetic effects, while also providing empirical support for the leash model of host control, which posits that bacterial communities are driven not only by bottom-up species interactions, but also by top-down host regulation. Taken together, our findings have broad implications for understanding host-microbe interactions as well as prokaryotic diversity in general. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Animal bodies harbour a complex and diverse community of microorganisms and accumulating evidence has revealed that microbes can influence the hosts’ behaviour, for example by altering body odours. Microbial communities produce odorant molecules as metabolic by-products and thereby modulate the biochemical signalling profiles of their animal hosts. As the diversity and the relative abundance of microbial species are influenced by several factors including host-specific factors, environmental factors and social interactions, there are substantial individual variations in the composition of microbial communities. In turn, the variations in microbial communities would consequently affect social and communicative behaviour by influencing recognition cues of the hosts. Therefore, microbiota studies have a great potential to expand our understanding of recognition of conspecifics, group members and kin. In this review, we aim to summarize existing knowledge of the factors influencing the microbial communities and the effect of microbiota on olfactory cue production and social and communicative behaviour. We concentrate on avian taxa, yet we also include recent research performed on non-avian species when necessary.

We here review the role of olfaction in the Zebra Finch, a key avian model species. We summarize the use of olfaction in social communication and in nonsocial contexts; its impact in addition to other senses throughout life; and how olfaction is involved in kin recognition, mate choice, and inbreeding avoidance. Afterward we explore the mechanisms of the olfactory signal and potential ways of signal production. We review olfactory signal production in avian species, the influences from the genome, and potential impact of skin microbes on scent production. This is followed by a paragraph on the perception and processing of the olfactory signal, from olfactory receptor genes, the olfactory bulb to the neural pathways in the songbird brain. Finally, we summarize several methods to examine olfaction in Zebra Finches on both, the behavioral level as well as on the chemical structural level. Based on the reviewed contents, we provide an outlook to future directions of each of the research fields to gather a further understanding of the use and the function as well as of the underlying mechanisms of olfaction in the Zebra Finch.

An animals' body is densely populated with bacteria. Although a large number of investigations on physiological microbial colonisation have emerged in recent years, our understanding of the composition, ecology and function of the microbiota remains incomplete. Here, we investigated whether songbirds have an individual-specific skin microbiome that is similar across different body regions. We collected skin microbe samples from three different bird species (Taeniopygia gutatta, Lonchura striata domestica and Stagonopleura gutatta) at two body locations (neck region, preen gland area). To characterise the skin microbes and compare the bacterial composition, we used high-throughput 16S rRNA amplicon sequencing. This method proved suitable for identifying the skin microbiome of birds, even though the bacterial load on the skin appeared to be relatively low. We found that across all species, the two evaluated skin areas of each individual harboured very similar microbial communities, indicative of an individual-specific skin microbiome. Despite experiencing the same environmental conditions and consuming the same diet, significant differences in the skin microbe composition were identified among the three species. The bird species differed both quantitatively and qualitatively regarding the observed bacterial taxa. Although each species harboured its own unique set of skin microbes, we identified a core skin microbiome among the studied species. As microbes are known to influence the host's body odour, our findings of an individual-specific skin microbiome might suggest that the skin microbiome in birds is involved in the odour production and could encode information on the host's genotype.

We describe an easy method to test odour detection and recognition in 1-d-old zebra finch hatchlings (Taeniopygia guttata). Day-old chicks beg in a stereotypical posture, which can be induced by directing gentle puffs of air from a plastic wash bottle near the face. We used this method to experimentally test whether begging duration of chicks was indicative of nest odour recognition. We manipulated the olfactory environment of 12 nests throughout incubation and hatching with either an artificial odour (orange oil) or with a neutral control (tap water). We then presented these two stimulus odours to 25 day-old chicks and measured the duration of the first begging bout exhibited for each odour. Zebra finches hatched in a nest environment enriched with orange oil scent begged significantly longer when exposed to orange oil odour, compared to control hatchlings. Our simple testing procedure can be used to efficiently quantify odour recognition and/or preference in altricial songbirds at a very early developmental stage.

In living color Animals live in a colorful world, but we rarely stop to think about how this color is produced and perceived, or how it evolved. Cuthill et al. review how color is used for social signals between individual animals and how it affects interactions with parasites, predators, and the physical environment. New approaches are elucidating aspects of animal coloration, from the requirements for complex cognition and perception mechanisms to the evolutionary dynamics surrounding its development and diversification. Science , this issue p. eaan0221

Our understanding of fundamental organismal biology has been disproportionately influenced by studies of a relatively small number of ‘model’ species extensively studied in captivity. Laboratory populations of model species are commonly subject to a number of forms of past and current selection that may affect experimental outcomes. Here, we examine these processes and their outcomes in one of the most widely used vertebrate species in the laboratory – the zebra finch (Taeniopygia guttata). This important model species is used for research across a broad range of fields, partly due to the ease with which it can be bred in captivity. However despite this perceived amenability, we demonstrate extensive variation in the success with which different laboratories and studies bred their subjects, and overall only 64% of all females that were given the opportunity, bred successfully in the laboratory. We identify and review several environmental, husbandry, life-history and behavioural factors that potentially contribute to this variation. The variation in reproductive success across individuals could lead to biases in experimental outcomes and drive some of the heterogeneity in research outcomes across studies. The zebra finch remains an excellent captive animal system and our aim is to sharpen the insight that future studies of this species can provide, both to our understanding of this species and also with respect to the reproduction of captive animals more widely. We hope to improve systematic reporting methods and that further investigation of the issues we raise will lead both to advances in our fundamental understanding of avian reproduction as well as to improvements in future welfare and experimental efficiency.

Environmental conditions experienced early in life have been shown to significantly affect growth trajectories at later stages in many vertebrate species. Amphibians typically have a biphasic life history, with an aquatic larval phase during early development and a subsequent terrestrial adult phase after completed metamorphosis. Thus, the early conditions have an especially strong impact on the future survival and fitness of amphibians. We studied whether early nutritional conditions affect the behavioural reaction of fire salamander larvae (Salamandra salamandra) before completion of metamorphosis. Fire salamander larvae reared under rich nutritional conditions were heavier and larger, displayed better body condition overall throughout the first three month of life and metamorphosed earlier compared with larvae raised under poor nutritional conditions. Specifically, we tested whether larvae reared under these different conditions differed with respect to their risk-taking behaviour and activity. We found no differences in the activity of larvae with respect to their experienced early food conditions. However, larvae reared under poor nutritional conditions hid significantly more often in a risk-taking test than larvae reared under rich food conditions. This increase in shelter-seeking behaviour might be an adaptation to reduce the risk of larval drift or an adaptation to compensate for physiological deficits in part by appropriate behavioural reactions. Our results indicate that environmental conditions, such as food availability, may lead to different behavioural strategies.

Finding a potential mate, that is an individual of the opposite sex, is a fundamental step for sexual reproduction in animal species. Signals involved in the context of mate attraction are mediated by acoustic, visual, and/or chemical signals. For amphibians in general, and especially for many newt and salamander species, chemical cues are known to play important roles in inter- and intraspecific communication. We therefore investigated the use of olfactory cues for sex recognition in terrestrial fire salamanders (Salamandra salamandra) that belong to the group of true salamanders within the Salamandridae. Specifically, we performed odor preference tests with adult salamanders of both sexes and tested whether substrate-borne chemical cues provide sex-specific signals. We found an overall preference for chemical cues of the opposite sex, i.e. males and females differed significantly in their preference for a specific sex. Females spent significantly more time in the compartment with the chemical cues of a male, whereas males did not show a significant preference, but in general more males preferred the compartment with the chemical cues of a female. Our results suggest that fire salamanders are capable to discriminate the sex of conspecific individuals based on pure chemical cues. We discuss our results in the light of the presence of dorsal glands in the males' cloaca of true salamander species, from which pheromones can be directly released onto the substrate.

Scent marks are relatively long-lived signals that can be perceived by conspecifics when the producer is absent. Therefore, it is often not obvious to whom the signal is directed. In daytime roosts of the polygynous greater sac-winged bat, males scent mark territories with facial gland secretions. Territories are a valuable resource for males, as they offer exclusive courtship opportunities, which results in increased male reproductive success and, consequently, increased male-male competition over territories. The information encoded in male scent marks could, therefore, be either directed at females as part of an olfactory courtship display or at male competitors as part of territorial behaviour. We expected territorial males to scent mark in the morning, shortly before females return to the territory and close to female roosting sites, if scent marks are directed at females as part of the courtship display. And we expected harem males to scent mark at the territory boundaries, where male-male encounters are most likely to occur, if scent marks are directed at male competitors. We found that males marked more frequently in the afternoon, at a time when all females have already left the territory, and harem males marked at the territory boundaries and not inside their territory in the area where females roost. At boundaries males fan volatiles from specialised wing sacs towards competitors outside the territory. Scent marking of male Saccopteryx bilineata might therefore be congruent with the assessment-hypothesis, which states that scent marks offer intruders the possibility to make an olfactory assessment of the territory owner without direct physical interaction. Thus, scent marks of male S. bilineata are most likely influenced by male-male competition and not by female choice.

Combining chemical analysis and odour preference tests, we asked whether two closely related sympatric species of sac-winged bats use odour for species recognition. Males of the two sister species Saccopteryx bilineata and Saccopteryx leptura have pouches containing an odoriferous liquid in their antebrachial wing membrane, which is used in S. bilineata during courtship displays. Although both species occasionally share the same daytime roosts and are morphologically similar, there is no evidence for interbreeding. We compared the production and composition of the wing sac odorant in male S. leptura and S. bilineata and performed odour preference tests with female S. bilineata. Similar to male S. bilineata, male S. leptura cleansed and refilled their wing sacs with secretions, but they spent more time each day in doing so than male S. bilineata. Chemical analysis by gas chromatography and mass spectrometry revealed that male Saccopteryx carried species-specific scents in their wing sacs. Binary choice tests confirmed that female S. bilineata preferred the wing sac scents of male S. bilineata to those of the sister species, suggesting that the species specificity of male wing sac scents maintain the pre-mating isolation barrier between these closely related species.

Like many other mammals, Saccopteryx bilineata exhibits a polygynous mating system, in which each male defends a group of females called a harem. Colonies consist of several harems, and nonharem males roost adjacent to harems. Unlike most other mammals, females disperse from their natal colony and most juvenile males remain in it. Thus, colonies consist of patrilines, which promotes intense local mate competition. Females are in estrus during a few weeks at the end of the rainy season. Mating is most likely initiated by females and preceded by intense courtship displays of males. Forty percent of colony males do not sire any offspring during their tenure in the colony, whereas a few males can sire up to 6 offspring in a single year. Males use olfactory, visual, and acoustic signals for courtship, and they demonstrate territory ownership by scent marks, lowfrequency calls, and visual displays. Harem males sire on average more offspring than do nonharem males but produce only 30% of the offspring within their own harem territory, with 70% being sired by other harem males or nonharem males. Reproductive success of males increases with decreasing size, fluctuating asymmetry, and fundamental frequency of territorial calls. In addition, females that are closely related to the harem holder are more likely to mate with other males than with the harem holder. Sexual selection in S. bilineata is most likely influenced by intense local mate competition caused by scarce roosts and the patrilineal organization of colonies.

At night, bats utter loud echolocation calls at high repetition rates that may reveal the location and current behaviour of callers to eavesdropping bats. Given the strong attenuation of echolocation calls, we predicted that territorial males of a harem-polygynous species ought to forage at close distance to females to survey their movements by social eavesdropping. We estimated a maximum detection distance of 38 m for echolocation calls of Saccopteryx bilineata (Temminck, 1838) (Chiroptera; Emballonuridae) broadcasted within the forest under the sound transmission conditions of our study site and for an assumed signal detection threshold of 20 dB SPL (50 m for a threshold of 0 dB SPL). We then simultaneously radio-tracked the nocturnal movements of eight male–female pairs that each rested in the same harem territory during the day and measured the distances at which dyads foraged. Male–female pairs foraged at a median distance of 139 m. In the case of 90% of simultaneous bearings, males foraged at distances that prevented eavesdropping on 44 kHz echolocation calls (>38 m; 87% of radio fixes >50 m). Males and females of the same daytime territory roosted, on average, 226 ± 194 m apart from each other at night. Thus, males were most likely unaware of where females foraged as a result of the strong attenuation of female echolocation calls. In general, such acoustic constraints on social eavesdropping may promote extra-harem group paternities, female choice, and sperm competition in bats, and may therefore present an important selective force in the evolution of bat mating systems.

Illuminating the ability of individuals to react to different selective forces caused by environmental differences is crucial to understand population divergence and speciation in the context of habitat adaptation. In a common environment experiment performed under standardised laboratory conditions, we examined whether female fire salamanders (Salamandra salamandra) originating from a recently adaptively diverged population displayed behavioural phenotype differences related to larval deposition behaviour. Females of this species typically deposit their larvae in small first-order streams; however, in our study population, females also use temporary ponds. These two habitat types display major ecological differences that strongly influence larval growth and survival. We observed that females differed in larval deposition behaviour and maternal investment. Pond-type females extended larval deposition over an increased time period and tended to exhibit more deposition events compared with stream-type females. Over successive deposition events, the body condition of larvae deposited by stream-type females decreased faster than that of larvae deposited by pond-type females. These differences in larval deposition behaviour may represent a bet-hedging strategy, given that ponds are more constrained in terms of desiccation and food availability than streams. The lengthened deposition period enabled pond-type females to deposit larger larvae towards the end of the deposition period, compared with stream-type females. Although the studied population only diverged recently, we observed significant behavioural differences between differentially adapted females, demonstrating the importance of behavioural differences in habitat adaptation in the context of speciation processes.

Chemical communication underpins virtually all aspects of vertebrate social life, yet remains poorly understood because of its highly complex mechanistic basis. We therefore used chemical fingerprinting of skin swabs and genetic analysis to explore the chemical cues that may underlie mother–offspring recognition in colonially breeding Antarctic fur seals. By sampling mother–offspring pairs from two different colonies, using a variety of statistical approaches and genotyping a large panel of microsatellite loci, we show that colony membership, mother–offspring similarity, heterozygosity, and genetic relatedness are all chemically encoded. Moreover, chemical similarity between mothers and offspring reflects a combination of genetic and environmental influences, the former partly encoded by substances resembling known pheromones. Our findings reveal the diversity of information contained within chemical fingerprints and have implications for understanding mother–offspring communication, kin recognition, and mate choice.

Although it has long been disregarded, the accumulating evidence suggests that birds use chemical cues for communication. However, the impact of olfaction on avian mate choice and its potential function in inbreeding avoidance remain unknown. We compared the mate choice and reproductive behaviour between females experimentally deprived of their sense of smell and intact control females. Each female was allowed to mate with two unfamiliar males, an unrelated male and a full brother. If olfactory cues are involved in kin recognition and/or mate choice, we expect the smelling females to reproduce with the unrelated males and the anosmic females to mate randomly with respect to relatedness. The paternity analysis revealed that the anosmic females reproduced randomly with respect to relatedness. Contrary to our expectations, the females with an intact olfactory mucosa demonstrated a decrease in reproduction; they produced significantly fewer eggs than the anosmic females, and the chicks in only one of the cages reached independence. Thus, the female's ability to smell in the presence of the unfamiliar brother had the unexpected effect of impairing the reproductive behaviour of the females. This is the first study to report directly the critical role of olfactory signals in reproduction and its potential function in preventing maladaptive breeding attempts with kin in female songbirds.