A genomic basis of vocal rhythm in birds.

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

Age-related changes in migratory behaviour within the first annual cycle of a passerine bird.

First time migrants (juveniles hereafter) of many species migrate without specific knowledge of non-breeding locations, but experience may aid adults in timing and route decisions because they can migrate more efficiently to their previous non-breeding sites. Consequently, we expect a transition to more efficient migratory behaviour with age, but when and how this happens is little known. We used light-level geolocator data from Cyprus wheatears Oenanthe cypriaca to compare migration timing and route directness between juveniles and adults, and repeatability of their timing and non-breeding locations. We predicted that juveniles would depart and arrive later than adults for both autumn and spring migration; that duration of migration would be greater for juveniles; that routes taken by juveniles would be less direct than those for adults; and that autumn and spring departure timing, and non-breeding locations, would be more repeatable for adults between two years than for juveniles between their first and subsequent migration. We found that juveniles departed significantly later than adults in autumn but there was no difference in arrival timing, and although spring departure timings did not differ, juveniles arrived on the breeding grounds later than adults. Nevertheless, we found no significant age-related difference in the duration of migration in autumn or spring. Yet, juvenile migrations were less direct than those of adults in autumn, but not spring. We found evidence that spring departure timing and non-breeding locations were repeatable for adults but not juveniles. Our findings show that age-related changes in migratory behaviour begin to occur during the first annual cycle demonstrating the potential for early adaptation to environmental variability within an individual's life.

Continent-wide patterns of song variation predicted by classical rules of biogeography.

Physiological constraints related to atmospheric temperature pose a limit to body and appendage size in endothermic animals. This relationship has been summarised by two classical principles of biogeography: Bergmann's and Allen's rules. Body size may also constrain other phenotypic traits important in ecology, evolution and behaviour, and such effects have seldom been investigated at a continental scale. Through a multilevel-modelling approach, we demonstrate that continent-wide morphology of related African barbets follows predictions of Bergmann's rule, and that body size mirrors variation in song pitch, an acoustic trait important in species recognition and sexual selection. Specifically, effects on song frequency in accordance with Bergmann's rule dwarf those of acoustic adaptation at a continental scale. Our findings suggest that macroecological patterns of body size can influence phenotypic traits important in ecology and evolution, and provide a baseline for further studies on the effects of environmental change on bird song.

First detection of WNV RNA presence in field-collected mosquitoes in Cyprus.

West Nile virus (WNV) infections have increased over recent years to the extent that WNV has become one of the most widespread arboviruses in the world, with potential consequences for both human and animal health. While much is known about WNV and the vectors that transmit it from their primary hosts across continental Europe, little is known about the epidemiology of the disease on the island of Cyprus. In this study, the aim was to investigate the prevalence of WNV infection in potential mosquito vectors for the first time in the Republic of Cyprus, using WNV surveillance of mosquitoes. Mosquitoes were collected in 2019, during which an outbreak in humans had occurred, and sampled mosquitoes were then examined for WNV infection by testing them for the presence of WNV RNA. Of 126 mosquito pools tested, one pool, containing Culex pipiens mosquitoes sampled from the Nicosia district, was found to be positive for the presence of WNV RNA. The positive pool found in this study represents the first demonstration of WNV in mosquitoes in Cyprus and confirms that human cases in Cyprus are likely the result of transmission via local Culex mosquitoes.

The potential function of post-fledging dispersal behavior in first breeding territory selection for males of a migratory bird.

One possible hypothesis for the function of post-fledging dispersal is to locate a suitable future breeding area. This post-fledging period may be particularly important in migratory species because they have a limited period to gather information prior to autumn migration, and in protandrous species, males must quickly acquire a territory after returning from spring migration to maximize their fitness. Here we use color-ring resightings to investigate how the post-fledging dispersal movements of the Cyprus wheatear Oenanthe cypriaca, a small migratory passerine, relate to their first breeding territory the following year when they return from migration. We found that males established first breeding territories that were significantly closer to their post-fledging location than to their natal sites or to post-fledging locations of other conspecifics, but these patterns were not apparent in females. Our findings suggest that familiarity with potential breeding sites may be important for juveniles of migratory species, particularly for the sex that acquires and advertises breeding territories. Exploratory dispersal prior to a migrant's first autumn migration may contribute toward its breeding success the following year, further highlighting the importance of early seasonal breeding on fitness and population dynamics more generally.

Evidence of West Nile virus seropositivity in wild birds on the island of Cyprus.

West Nile Virus (WNV)1 is an emerging pathogen in Cyprus, with the first human case of infection reported in 2016, and another documented in 2018. A cluster of cases in humans was then reported in 2019. However, little is known regarding which avian species might bring WNV to Cyprus. Here, we investigated seroprevalence of WNV antibodies in migratory and resident birds, captured across Cyprus to assess to what extent human populations might be exposed to WNV. We used Enzyme-Linked Immunosorbent Assay (ELISA)2 to test for the presence of WNV antibodies in 836 avian blood samples of 44 species captured between 2015 and 2020. A seropositivity rate of 1.3 % was found. The majority of seropositive wild birds belonged to the migratory species Sylvia atricapilla, a common and widespread migrant, implying a high risk of WNV being introduced throughout Cyprus.

CYP2J19 mediates carotenoid colour introgression across a natural avian hybrid zone.

It has long been of interest to identify the phenotypic traits that mediate reproductive isolation between related species, and more recently, the genes that underpin them. Much work has focused on identifying genes associated with animal colour, with the candidate gene CYP2J19 identified in laboratory studies as the ketolase converting yellow dietary carotenoids to red ketocarotenoids in birds with red pigments. However, evidence that CYP2J19 explains variation between red and yellow feather coloration in wild populations of birds is lacking. Hybrid zones provide the opportunity to identify genes associated with specific traits. Here we investigate genomic regions associated with colour in red-fronted and yellow-fronted tinkerbirds across a hybrid zone in southern Africa. We sampled 85 individuals, measuring spectral reflectance of forecrown feathers and scoring colours from photographs, while testing for carotenoid presence with Raman spectroscopy. We performed a genome-wide association study to identify associations with carotenoid-based coloration, using double-digest RAD sequencing aligned to a short-read whole genome of a Pogoniulus tinkerbird. Admixture mapping using 104,933 single nucleotide polymorphisms (SNPs) identified a region of chromosome 8 that includes CYP2J19 as the only locus with more than two SNPs significantly associated with both crown hue and crown score, while Raman spectra provided evidence of ketocarotenoids in red feathers. Asymmetric backcrossing in the hybrid zone suggests that yellow-fronted females mate more often with red-fronted males than vice versa. Female red-fronted tinkerbirds mating assortatively with red-crowned males is consistent with the hypothesis that converted carotenoids are an honest signal of quality.

Tape lures swell bycatch on a Mediterranean island harbouring illegal bird trapping.

Mediterranean islands provide shelter and sustenance for millions of migrating birds each year. Humans have historically exploited bird migration through hunting. In Cyprus, trapping birds during their migratory peak is considered a tradition, but has long been against the law. Illegal bird trapping is lucrative, however, with trappers using tape lures that broadcast birdsong to increase capture rates. It results in the slaughter of millions of birds each year. Yet, scientific studies quantifying capture rates of target and nontarget species using methods employed by trappers are lacking. Here, we show using playback experiments that tape lures lead to an order of magnitude greater capture rates of target species, but also significantly increase bycatch, which may include species of conservation concern. Conservation efforts focusing on minimizing illegal bird killing should also consider tape lures and their contribution to the overall impact of trapping on avian populations.

Habitat availability influences migration speed, refueling patterns and seasonal flyways of a fly-and-forage migrant.

BACKGROUND: Despite our understanding of the principal factors that shape bird migration strategies, there is conflicting evidence regarding the role of habitat in shaping migration routes and schedules, including day and night activity and differences between autumn and spring. For fly-and-forage migrants, we predict that habitat characteristics might guide migration speed, route selection and migrating schedules. METHODS: We use solar-powered GPS transmitters, obtaining high accuracy data, to monitor the migratory movements of Eleonora's falcon breeding in Cyprus, which is the easternmost breeding population of the species. We tested for potential preferences in habitat characteristics along the migration routes, separately for the northern, drier part and the more vegetated southern part of the trips. We also examined the relationship between migration speed and vegetative cover during day and at night, accounting for wind support. RESULTS: We found that tagged individuals repeatedly exhibited an anticlockwise loop migration pattern with spring routes being more easterly than autumn ones. We identified a preference for migration through vegetation-rich areas, where during daytime tagged individuals travel at slower migration speeds compared to vegetation-poor areas, indicating fly-and-forage activity. Birds roosted during most nights, combining refueling stopovers at selected vegetation-rich areas before or after crossing ecological barriers. Conversely, both during day and night, tagged individuals overflew unsuitable habitats more quickly. CONCLUSIONS: Our results suggest that habitat is an important factor in Eleonora's falcon migratory strategies. Active selection of vegetation rich areas in combination with reduced migration speeds there, allows the migrating falcons to combine migration during the day with fly-and-forage refueling, while roosting most nights.

Range-wide spatial mapping reveals convergent character displacement of bird song.

A long-held view in evolutionary biology is that character displacement generates divergent phenotypes in closely related coexisting species to avoid the costs of hybridization or ecological competition, whereas an alternative possibility is that signals of dominance or aggression may instead converge to facilitate coexistence among ecological competitors. Although this counterintuitive process-termed convergent agonistic character displacement-is supported by recent theoretical and empirical studies, the extent to which it drives spatial patterns of trait evolution at continental scales remains unclear. By modelling the variation in song structure of two ecologically similar species of Hypocnemis antbird across western Amazonia, we show that their territorial signals converge such that trait similarity peaks in the sympatric zone, where intense interspecific territoriality between these taxa has previously been demonstrated. We also use remote sensing data to show that signal convergence is not explained by environmental gradients and is thus unlikely to evolve by sensory drive (i.e. acoustic adaptation to the sound transmission properties of habitats). Our results suggest that agonistic character displacement driven by interspecific competition can generate spatial patterns opposite to those predicted by classic character displacement theory, and highlight the potential role of social selection in shaping geographical variation in signal phenotypes of ecological competitors.

Rapid song divergence leads to discordance between genetic distance and phenotypic characters important in reproductive isolation.

The criteria for species delimitation in birds have long been debated, and several recent studies have proposed new methods for such delimitation. On one side, there is a large consensus of investigators who believe that the only evidence that can be used to delimit species is molecular phylogenetics, and with increasing numbers of markers to gain better support, whereas on the other, there are investigators adopting alternative approaches based largely on phenotypic differences, including in morphology and communication signals. Yet, these methods have little to say about rapid differentiation in specific traits shown to be important in reproductive isolation. Here, we examine variation in phenotypic (morphology, plumage, and song) and genotypic (mitochondrial and nuclear DNA) traits among populations of yellow-rumped tinkerbird Pogoniulus bilineatus in East Africa. Strikingly, song divergence between the P. b. fischeri subspecies from Kenya and Zanzibar and P. b. bilineatus from Tanzania is discordant with genetic distance, having occurred over a short time frame, and playback experiments show that adjacent populations of P. b. bilineatus and P. b. fischeri do not recognize one another's songs. While such rapid divergence might suggest a founder effect following invasion of Zanzibar, molecular evidence suggests otherwise, with insular P. b. fischeri nested within mainland P. b. fischeri. Populations from the Eastern Arc Mountains are genetically more distant, yet share the same song with P. b. bilineatus from Coastal Tanzania and Southern Africa, suggesting they would interbreed. We believe investigators ought to examine potentially rapid divergence in traits important in species recognition and sexual selection when delimiting species, rather than relying entirely on arbitrary quantitative characters or molecular markers.

Current and future suitability of wintering grounds for a long-distance migratory raptor.

Conservation of migratory species faces the challenge of understanding the ecological requirements of individuals living in two geographically separated regions. In some cases, the entire population of widely distributed species congregates at relatively small wintering areas and hence, these areas become a priority for the species' conservation. Satellite telemetry allows fine tracking of animal movements and distribution in those less known, often remote areas. Through integrating satellite and GPS data from five separated populations comprising most of the breeding range, we created a wide habitat suitability model for the Eleonora's falcon on its wintering grounds in Madagascar. On this basis, we further investigated, for the first time, the impact of climate change on the future suitability of the species' wintering areas. Eleonora's falcons are mainly distributed in the north and along the east of Madagascar, exhibiting strong site fidelity over years. The current species' distribution pattern is associated with climatic factors, which are likely related to food availability. The extent of suitable areas for Eleonora's falcon is expected to increase in the future. The integration of habitat use information and climatic projections may provide insights on the consequences of global environmental changes for the long-term persistence of migratory species populations.

Smithornis broadbills produce loud wing song by aeroelastic flutter of medial primary wing feathers.

Broadbills in the genus Smithornis produce a loud brreeeeet during a distinctive flight display. It has been posited that this klaxon-like sound is generated non-vocally with the outer wing feathers (P9, P10), but no scientific studies have previously addressed this hypothesis. Although most birds that make non-vocal communication sounds have feathers with a shape distinctively modified for sound production, Smithornis broadbills do not. We investigated whether this song is produced vocally or with the wings in rufous-sided broadbill (S. rufolateralis) and African broad bill (S. capensis). In support of the wing song hypothesis, synchronized high-speed video and sound recordings of displays demonstrated that sound pulses were produced during the downstroke, subtle gaps sometimes appeared between the outer primary feathers P6-P10, and wing tip speed reached 16 m s(-1) Tests of a spread wing in a wind tunnel demonstrated that at a specific orientation, P6 and P7 flutter and produce sound. Wind tunnel tests on individual feathers P5-P10 from a male of each species revealed that while all of these feathers can produce sound via aeroelastic flutter, P6 and P7 produce the loudest sounds, which are similar in frequency to the wing song, at airspeeds achievable by the wing tip during display flight. Consistent with the wind tunnel experiments, field manipulations of P6, P7 and P8 changed the timbre of the wing song, and reduced its tonality, demonstrating that P6 and P7 are together the sound source, and not P9 or P10. The resultant wing song appears to have functionally replaced vocal song.

Predicting bird song from space.

Environmentally imposed selection pressures are well known to shape animal signals. Changes in these signals can result in recognition mismatches between individuals living in different habitats, leading to reproductive divergence and speciation. For example, numerous studies have shown that differences in avian song may be a potent prezygotic isolating mechanism. Typically, however, detailed studies of environmental pressures on variation in animal behavior have been conducted only at small spatial scales. Here, we use remote-sensing data to predict animal behavior, in this case, bird song, across vast spatial scales. We use remotely sensed data to predict the song characteristics of the little greenbul (Andropadus virens), a widely distributed African passerine, found across secondary and mature rainforest habitats and the rainforest-savanna ecotone. Satellite data that captured ecosystem structure and function explained up to 66% of the variation in song characteristics. Song differences observed across habitats, including those between human-altered and mature rainforest, have the potential to lead to reproductive divergence, and highlight the impacts that both natural and anthropogenic change may have on natural populations. Our approach offers a novel means to examine the ecological correlates of animal behavior across large geographic areas with potential applications to both evolutionary and conservation biology.

The evolutionary consequences of interspecific aggression.

Competition has always been a cornerstone of evolutionary biology, and aggression is the predominant form of direct competition in animals, but the evolutionary effects of aggression between species are curiously understudied. Only in the past few years, existing theoretical frameworks have been extended to include interspecific aggression, and significant empirical advances have been made. After arguing that agonistic character displacement (ACD) theory provides the most suitable theoretical framework, we review new empirical evidence for ACD and the results of mathematical models of the process. We consider how ACD can be distinguished empirically from ecological and reproductive character displacement and the additional challenges posed by developmental plasticity. We also provide the first taxonomically broad review of theoretical and empirical work on the effects of interspecific aggression on species coexistence and range limits. We conclude by highlighting promising directions for future research on the evolutionary effects of interspecific aggression.

Testing alternative hypotheses for evolutionary diversification in an African songbird: rainforest refugia versus ecological gradients.

Geographic isolation in rainforest refugia and local adaptation to ecological gradients may both be important drivers of evolutionary diversification. However, their relative importance and the underlying mechanisms of these processes remain poorly understood because few empirical studies address both putative processes in a single system. A key question is to what extent is divergence in signals that are important in mate and species recognition driven by isolation in rainforest refugia or by divergent selection across ecological gradients? We studied the little greenbul, Andropadus virens, an African songbird, in Cameroon and Uganda, to determine whether refugial isolation or ecological gradients better explain existing song variation. We then tested whether song variation attributable to refugial or ecological divergence was biologically meaningful using reciprocal playback experiments to territorial males. We found that much of the existing song variation can be explained by both geographic isolation and ecological gradients, but that divergence across the gradient, and not geographic isolation, affects male response levels. These data suggest that ecologically divergent traits, independent of historical isolation during glacial cycles, can promote reproductive isolation. Our study provides further support for the importance of ecology in explaining patterns of evolutionary diversification in ecologically diverse regions of the planet.

Acoustic localization of antbirds in a Mexican rainforest using a wireless sensor network.

Acoustic localization is a promising method to passively observe vocal animal species, but remains difficult and time consuming to employ. To reduce the labor intensity and impact of deployment, an acoustic localization system has been developed consisting of battery powered wireless sensor nodes. The system also has the ability to perform an acoustic self-survey, which compares favorably in accuracy to global positioning system survey methods, especially in environments such as forest. The self-survey and localization accuracy of the system was tested in the neotropical rainforest of Chiapas, Mexico. A straight-forward and robust correlation sum localization computation method was utilized and is described in detail. Both free-ranging wild antbird songs and songs played from a speaker were localized with mean errors of 0.199 m and 0.445 m, respectively. Finally, additional tests utilizing only a short segment of each song or a subset of sensor nodes were performed and found to minimally affect localization accuracy. The use of a wireless sensor network for acoustic localization of animal vocalizations offers greater ease and flexibility of deployment than wired microphone arrays without sacrificing accuracy.

Character displacement of song and morphology in African tinkerbirds.

Divergence in acoustic signals between populations of animals can lead to species recognition failure, reproductive isolation, and speciation. Character displacement may facilitate coexistence of species in natural communities, yet evidence for character displacement in acoustic signals is scant. Here, we find evidence of character displacement in song as well as body size and bill size of 2 related African tinkerbirds. Playback experiments indicate that related species' songs are perceived differently in sympatry than in allopatry. We suggest character displacement occurs in phenotypic traits facilitating species recognition, which has important implications for understanding the processes that lead to speciation and diversification. Because many of the sites where the 2 species coexist are areas where pristine rainforest has been degraded, results also suggest that anthropogenic pressures resulting from deforestation may be a contributing cause of character displacement in these species.

Automated species recognition of antbirds in a Mexican rainforest using hidden Markov models.

Behavioral and ecological studies would benefit from the ability to automatically identify species from acoustic recordings. The work presented in this article explores the ability of hidden Markov models to distinguish songs from five species of antbirds that share the same territory in a rainforest environment in Mexico. When only clean recordings were used, species recognition was nearly perfect, 99.5%. With noisy recordings, performance was lower but generally exceeding 90%. Besides the quality of the recordings, performance has been found to be heavily influenced by a multitude of factors, such as the size of the training set, the feature extraction method used, and number of states in the Markov model. In general, training with noisier data also improved recognition in test recordings, because of an increased ability to generalize. Considerations for improving performance, including beamforming with sensor arrays and design of preprocessing methods particularly suited for bird songs, are discussed. Combining sensor network technology with effective event detection and species identification algorithms will enable observation of species interactions at a spatial and temporal resolution that is simply impossible with current tools. Analysis of animal behavior through real-time tracking of individuals and recording of large amounts of data with embedded devices in remote locations is thus a realistic goal.