The Enrichment of Specific Hair Follicle-Associated Cell Populations in Plucked Hairs Offers an Opportunity to Study Gene Expression Underlying Hair Traits.

Gene expression differences can assist in characterizing important underlying genetic mechanisms between different phenotypic traits. However, when population-dense tissues are studied, the signals from scarce populations are diluted. Therefore, appropriately choosing a sample collection method that enriches a particular type of effector cells might yield more specific results. To address this issue, we performed a polyA-selected RNA-seq experiment of domestic horse (Equus ferus caballus) plucked-hair samples and skin biopsies. Then, we layered the horse gene abundance results against cell type-specific marker genes generated from a scRNA-seq supported with spatial mapping of laboratory mouse (Mus musculus) skin to identify the captured populations. The hair-plucking and skin-biopsy sample-collection methods yielded comparable quality and quantity of RNA-seq results. Keratin-related genes, such as KRT84 and KRT75, were among the genes that showed higher abundance in plucked hairs, while genes involved in cellular processes and enzymatic activities, such as MGST1, had higher abundance in skin biopsies. We found an enrichment of hair-follicle keratinocytes in plucked hairs, but detected an enrichment of other populations, including epidermis keratinocytes, in skin biopsies. In mammalian models, biopsies are often the method of choice for a plethora of gene expression studies and to our knowledge, this is a novel study that compares the cell-type enrichment between the non-invasive hair-plucking and the invasive skin-biopsy sample-collection methods. Here, we show that the non-invasive and ethically uncontroversial plucked-hair method is recommended depending on the research question. In conclusion, our study will allow downstream -omics approaches to better understand integumentary conditions in both health and disease in horses as well as other mammals.

A QTL for conformation of back and croup influences lateral gait quality in Icelandic horses.

BACKGROUND: The back plays a vital role in horse locomotion, where the spine functions as a spring during the stride cycle. A complex interaction between the spine and the muscles of the back contribute to locomotion soundness, gait ability, and performance of riding and racehorses. Conformation is commonly used to select horses for breeding and performance in multiple horse breeds, where the back and croup conformation plays a significant role. The conformation of back and croup plays an important role on riding ability in Icelandic horses. However, the genes behind this trait are still unknown. Therefore, the aim of this study was to identify genomic regions associated with conformation of back and croup in Icelandic horses and to investigate their effects on riding ability. One hundred seventy-seven assessed Icelandic horses were included in the study. A genome-wide association analysis was performed using the 670 K+ Axiom Equine Genotyping Array, and the effects of different haplotypes in the top associated region were estimated for riding ability and additional conformation traits assessed during breeding field tests. RESULTS: A suggestive quantitative trait loci (QTL) for the score of back and croup was detected on Equus caballus (ECA) 22 (p-value = 2.67 × 10- 7). Haplotype analysis revealed two opposite haplotypes, which resulted in higher and lower scores of the back and croup, respectively (p-value < 0.001). Horses with the favorable haplotype were more inclined to have a well-balanced backline with an uphill conformation and had, on average, higher scores for the lateral gaits tölt (p-value = 0.02) and pace (p-value = 0.004). This genomic region harbors three genes: C20orf85, ANKRD60 and LOC100056167. ANKRD60 is associated with body height in humans. C20orf85 and ANKRD60 are potentially linked to adolescent idiopathic scoliosis in humans. CONCLUSIONS: Our results show that the detected QTL for conformation of back and croup is of importance for quality of lateral gaits in Icelandic horses. These findings could result in a genetic test to aid in the selection of breeding horses, thus they are of major interest for horse breeders. The results may also offer a gateway to comparative functional genomics by potentially linking both motor laterality and back inclination in horses with scoliosis in humans.

Contrasting stripes are a widespread feature of group living in birds, mammals and fishes.

Grouping is a widespread form of predator defence, with individuals in groups often performing evasive collective movements in response to attack by predators. Individuals in these groups use behavioural rules to coordinate their movements, with visual cues about neighbours' positions and orientations often informing movement decisions. Although the exact visual cues individuals use to coordinate their movements with neighbours have not yet been decoded, some studies have suggested that stripes, lines, or other body patterns may act as conspicuous conveyors of movement information that could promote coordinated group movement, or promote dazzle camouflage, thereby confusing predators. We used phylogenetic logistic regressions to test whether the contrasting achromatic stripes present in four different taxa vulnerable to predation, including species within two orders of birds (Anseriformes and Charadriiformes), a suborder of Artiodactyla (the ruminants), and several orders of marine fishes (predominantly Perciformes) were associated with group living. Contrasting patterns were significantly more prevalent in social species, and tended to be absent in solitary species or species less vulnerable to predation. We suggest that stripes taking the form of light-coloured lines on dark backgrounds, or vice versa, provide a widespread mechanism across taxa that either serves to inform conspecifics of neighbours' movements, or to confuse predators, when moving in groups. Because detection and processing of patterns and of motion in the visual channel is essentially colour-blind, diverse animal taxa with widely different vision systems (including mono-, di-, tri-, and tetrachromats) appear to have converged on a similar use of achromatic patterns, as would be expected given signal-detection theory. This hypothesis would explain the convergent evolution of conspicuous achromatic patterns as an antipredator mechanism in numerous vertebrate species.

Solar and terrestrial radiations explain continental-scale variation in bird pigmentation.

Animals living on the earth's surface are protected from the damaging effects of solar ultraviolet (UV) radiation by melanin pigments that color their integument. UV levels that reach the earth's surface vary spatially, but the role of UV exposure in shaping clinal variations in animal pigmentation has never been tested. Here, we show at a continental scale in Europe that golden eagles Aquila chrysaetos reared in territories with a high solar UV-B radiation exposure deposit lower amounts of the sulphurated form of melanin (pheomelanin) in feathers and consequently develop darker plumage phenotypes than eagles from territories with lower radiation exposure. This clinal variation in pigmentation is also explained by terrestrial γ radiation levels in the rearing territories by a similar effect on the pheomelanin content of feathers, unveiling natural radioactivity as a previously unsuspected factor shaping animal pigmentation. These findings show for the first time the potential of solar and terrestrial radiations to explain pigmentation phenotype diversity in animals, including humans, at large spatial scales.

Melanins in Fossil Animals: Is It Possible to Infer Life History Traits from the Coloration of Extinct Species?

Paleo-colour scientists have recently made the transition from describing melanin-based colouration in fossil specimens to inferring life-history traits of the species involved. Two such cases correspond to counter-shaded dinosaurs: dark-coloured due to melanins dorsally, and light-coloured ventrally. We believe that colour reconstruction of fossils based on the shape of preserved microstructures-the majority of paleo-colour studies involve melanin granules-is not without risks. In addition, animals with contrasting dorso-ventral colouration may be under different selection pressures beyond the need for camouflage, including, for instance, visual communication or ultraviolet (UV) protection. Melanin production is costly, and animals may invest less in areas of the integument where pigments are less needed. In addition, melanocytes exposed to UV radiation produce more melanin than unexposed melanocytes. Pigment economization may thus explain the colour pattern of some counter-shaded animals, including extinct species. Even in well-studied extant species, their diversity of hues and patterns is far from being understood; inferring colours and their functions in species only known from one or few specimens from the fossil record should be exerted with special prudence.

Intraspecific eye color variability in birds and mammals: a recent evolutionary event exclusive to humans and domestic animals.

BACKGROUND: Human populations and breeds of domestic animals are composed of individuals with a multiplicity of eye (= iris) colorations. Some wild birds and mammals may have intraspecific eye color variability, but this variation seems to be due to the developmental stage of the individual, its breeding status, and/or sexual dimorphism. In other words, eye colour tends to be a species-specific trait in wild animals, and the exceptions are species in which individuals of the same age group or gender all develop the same eye colour. Domestic animals, by definition, include bird and mammal species artificially selected by humans in the last few thousand years. Humans themselves may have acquired a diverse palette of eye colors, likewise in recent evolutionary time, in the Mesolithic or in the Upper Paleolithic. PRESENTATION OF THE HYPOTHESIS: We posit two previously unrecognized hypotheses regarding eye color variation: 1) eye coloration in wild animals of every species tends to be a fixed trait. 2) Humans and domestic animal populations, on the contrary, have eyes of multiple colors. Sexual selection has been invoked for eye color variation in humans, but this selection mode does not easily apply in domestic animals, where matings are controlled by the human breeder. TESTING THE HYPOTHESIS: Eye coloration is polygenic in humans. We wish to investigate the genetics of eye color in other animals, as well as the ecological correlates. IMPLICATIONS OF THE HYPOTHESIS: Investigating the origin and function of eye colors will shed light on the reason why some species may have either light-colored irises (e.g., white, yellow or light blue) or dark ones (dark red, brown or black). The causes behind the vast array of eye colors across taxa have never been thoroughly investigated, but it may well be that all Darwinian selection processes are at work: sexual selection in humans, artificial selection for domestic animals, and natural selection (mainly) for wild animals.

Seabird mortality induced by land-based artificial lights.

Artificial lights at night cause high mortality of seabirds, one of the most endangered groups of birds globally. Fledglings of burrow-nesting seabirds, and to a lesser extent adults, are attracted to and then grounded (i.e., forced to land) by lights when they fly at night. We reviewed the current state of knowledge of seabird attraction to light to identify information gaps and propose measures to address the problem. Although species in families such as Alcidae and Anatidae can be grounded by artificial light, the most affected seabirds are petrels and shearwaters (Procellariiformes). At least 56 species of Procellariiformes, more than one-third of them (24) threatened, are subject to grounding by lights. Seabirds grounded by lights have been found worldwide, mainly on oceanic islands but also at some continental locations. Petrel breeding grounds confined to formerly uninhabited islands are particularly at risk from light pollution due to tourism and urban sprawl. Where it is impractical to ban external lights, rescue programs of grounded birds offer the most immediate and employed mitigation to reduce the rate of light-induced mortality and save thousands of birds every year. These programs also provide useful information for seabird management. However, these data are typically fragmentary, biased, and uncertain and can lead to inaccurate impact estimates and poor understanding of the phenomenon of seabird attraction to lights. We believe the most urgently needed actions to mitigate and understand light-induced mortality of seabirds are estimation of mortality and effects on populations; determination of threshold light levels and safe distances from light sources; documentation of the fate of rescued birds; improvement of rescue campaigns, particularly in terms of increasing recovery rates and level of care; and research on seabird-friendly lights to reduce attraction.

New Insights into the Cosmetic Behaviour of Bearded Vultures: Ferruginous Springs Are Shared Sequentially.

Different hypotheses have been proposed to explain the function of cosmetic behaviour of bearded vultures, being the signalling individual dominance status the most accepted. However, no advances have been made in understanding this mysterious behaviour, in part due to the secrecy of this species. With the help of camera traps and GPS devices we monitored the use of a ferruginous spring in the Pyrenees (Spain) providing new insights into this aspect of their behavioural ecology. Most of the visits (93.5%) involved a single bearded vulture and bathing behaviour only occurred when a single individual was present, confirming their secretive behaviour. A total of 50% of individuals that visited the site were non-adults, suggesting that cosmetic coloration functions as an attenuating signal that may also benefit subordinate individuals. Future studies with the help of new technologies could help to disentangle some questions about the real function of cosmetic coloration and their social relevance.

Generation of raptor diversity in Europe: linking speciation with climate changes and the ability to migrate.

Europe holds a rich community of diurnal birds of prey, and the highest proportion of transcontinental migratory raptorial species of any landmass. This study will test the hypotheses that the high diversification of the raptor assemblage in Europe is a recent event, that closely related species sharing the same trophic niches can only coexist in sympatry during the breeding period, when food availability is higher, and finally that migration is a function of size, with the smaller species in every trophic group moving further. A consensus molecular phylogeny for the 38 regular breeding species of raptors in Europe was obtained from BirdTree (www.birdtree.org). For the same species, a trophic niche cluster dendrogram was constructed. Size and migratory strategy were introduced in the resulting phylogeny, where trophic groups were also identified. Multispecific trophic groups tended to be composed of reciprocal sister species of different sizes, while monospecific groups (n = 3) were composed of highly specialized species. Many speciation events took place recently, during the glacial cycles of the Quaternary, and size divergence among competing species may be due to character displacement. Nowadays, the smaller species in every trophic group migrate to sub-Saharan Africa. This investigation illustrates how the rich assemblage of diurnal birds of prey in Europe, more diverse and more migratory than, for instance, the North American assemblage at equivalent latitudes, has emerged recently due to the multiplication of look-alike species with similar trophic ecologies, possibly in climate refugia during cold periods.

The Dynamics of Lepus granatensis and Oryctolagus cuniculus in a Mediterranean Agrarian Area: Are Hares Segregating from Rabbit Habitats after Disease Impact?

The genera Oryctolagus and Lepus (order Lagomorpha) are essential elements in the trophic chain in the Iberian Peninsula, being the main prey of many predators, including some highly endangered predators such as the Iberian lynx (Lynx pardinus). Myxomatosis, a disease producing tumorations in conjunctive tissues, and produced by the Myxoma Virus, has caused mass mortalities in rabbits (Oryctolagus cuniculus) for decades. Recently, the virus has jumped interspecifically from rabbits to hares, and this has created a depletion in hare populations, generating great concern. We analyzed the population dynamics and distribution of both lagomorph species in a Mediterranean agricultural area of the south of Spain since the 1990s with a combination of systematic and non-systematic data. The appearance of the outbreak in the Iberian hare (Lepus granatenis) in 2018 enabled us to undertake an opportunistic analysis of its effects on the spatial structure and assemblages, as well as on the niches of both species using PCA analyses and ordination techniques. Analysis of the mortality effect on daily and seasonal cycles was also conducted, and relations with the temporal dimension was tested using generalized lineal models (GLMs). In our results, in addition to population and temporal patterns, we could observe a restructuring in hare distribution after the mortality event, highlighting that prior to the outbreak, rabbit and hare populations were spatially differentiated, although with some overlaps and niche similarities. However, since the outbreak, hare populations have been excluded from rabbit areas, suggesting that in the absence of rabbits, the virus has more difficulties to infect hares. We also provide an overview of the effect of this population depletion on the ecological and socio-economic dimension of this region, pointing out the importance of this situation for the area.

Owl-like plaques of the Copper Age and the involvement of children.

In the Copper Age, slate engraved plaques were produced massively in the southwestern corner of the Iberian Peninsula. Researchers have speculated about the function of these palm-sized stone objects for more than a century, although most have favored the idea that they represented goddesses, and served ritual purposes. The plaques are engraved with different designs of varying complexity. In some of them, the ones sporting two large frontal eyes, we clearly see owls modelled after two species present in the area: the little owl (Athene noctua), and the long-eared owl (Asio otus). These two species, living in semi-open habitats, were possibly the most abundant owls around the human settlements and surrounding cultivated fields of the Chalcolithic period. People must have been aware of the owl presence and possibly interacted with them. Why owls but no other animals have been the models may relate to the fact they are the most anthropomorphic of all animals, with large frontally-placed eyes in their enormous heads. In the iconography, owls are systematically represented, even today, with their two eyes staring at the observer, as opposed to the lateral view used for any other animal. Additionally, slate is one of the commonest surface rocks in southwestern Iberia, and it provides a blank canvas for engraving lines using pointed tools made of flint, quartz or copper. The way slates exfoliate makes easy to craft owl-looking plaques. To silhouette animals other than owls in a recognizable way would request extra carving abilities and specific tools. Plaque manufacture and design were simple and did not demand high skills nor intensive labor as demonstrated in replication experiments. Owl engravings could have been executed by youngsters, as they resemble owls painted today by elementary school students. This also suggests that schematic drawings are universal and timeless. We propose that the owl-like slate plaques are the remains of a set of objects used in both playful activities and in ritual ceremonies. The actual engraving of the plaques may have been part of the game. Owlish slate plaques were often perforated twice at the top. We interpret this as insertion points for actual bird feathers added to the plaques, right at the place where tufts emerge in live owls. The frontier among play and ritual is diffuse in liminal societies and there is no contradiction in playing with animal-like toys and, at some point, using them as offerings as part of community rituals related, for instance, to the colossal megalithic tombs so characteristic of the Copper Age.

Fine-tuning the space, time, and host distribution of mycobacteria in wildlife.

BACKGROUND: We describe the diversity of two kinds of mycobacteria isolates, environmental mycobacteria and Mycobacterium bovis collected from wild boar, fallow deer, red deer and cattle in Doñana National Park (DNP, Spain), analyzing their association with temporal, spatial and environmental factors. RESULTS: High diversity of environmental mycobacteria species and M. bovis typing patterns (TPs) were found. When assessing the factors underlying the presence of the most common types of both environmental mycobacteria and M. bovis TPs in DNP, we evidenced (i) host species differences in the occurrence, (ii) spatial structuration and (iii) differences in the degree of spatial association of specific types between host species. Co-infection of a single host by two M. bovis TPs occurred in all three wild ungulate species. In wild boar and red deer, isolation of one group of mycobacteria occurred more frequently in individuals not infected by the other group. While only three TPs were detected in wildlife between 1998 and 2003, up to 8 different ones were found during 2006-2007. The opposite was observed in cattle. Belonging to an M. bovis-infected social group was a significant risk factor for mycobacterial infection in red deer and wild boar, but not for fallow deer. M. bovis TPs were usually found closer to water marshland than MOTT. CONCLUSIONS: The diversity of mycobacteria described herein is indicative of multiple introduction events and a complex multi-host and multi-pathogen epidemiology in DNP. Significant changes in the mycobacterial isolate community may have taken place, even in a short time period (1998 to 2007). Aspects of host social organization should be taken into account in wildlife epidemiology. Wildlife in DNP is frequently exposed to different species of non-tuberculous, environmental mycobacteria, which could interact with the immune response to pathogenic mycobacteria, although the effects are unknown. This research highlights the suitability of molecular typing for surveys at small spatial and temporal scales.

Sources of variation for nutritional condition indices of the plasma of migratory lesser kestrels in the breeding grounds.

Although published information on reference values for biochemical parameters in birds of prey has increased during the last years, little is known on their sources of variation. We used an insectivorous and small migratory raptor species, the lesser kestrel Falco naumanni, as a model. We looked for sources of variation of nutritional biochemical parameters (i.e. triglycerides, cholesterol, uric acid and urea) of both nestlings and adults. Reference values indicated that, as a rule, lesser kestrel showed more elevated triglycerides, urea and uric acid levels than other raptors. All analyzed factors except gender (i.e. year, colony, sampling time, presence/absence of a geolocator, body mass, laying date and capture date) reached significance for at least one biochemical parameter. In the morning, we found an important postprandial increase in the concentration of all biochemical parameters in nestlings, and uric acid and urea levels in adults. A positive relationship was detected between triglycerides and body mass in nestlings and adults. Although we did not find differences between blood biochemical parameters of the oldest and youngest chick of each brood, we found that cholesterol levels were lower in nestlings from larger broods. Coloration of tarsi (measured as brightness) was related to triglycerides and urea levels of nestlings and adults, respectively. The feeding habits of lesser kestrel probably explain the different levels and patterns of variation of metabolites in comparison to more carnivorous raptors eating mammals or birds.

MHC diversity and differential exposure to pathogens in kestrels (Aves: Falconidae).

Pathogen diversity is thought to drive major histocompatibility complex (MHC) polymorphism given that host's immune repertories are dependent on antigen recognition capabilities. Here, we surveyed an extensive community of pathogens (n = 35 taxa) and MHC diversity in mainland versus island subspecies of the Eurasian kestrel Falco tinnunculus and in a sympatric mainland population of the phylogenetically related lesser kestrel Falco naumanni. Insular subspecies are commonly exposed to impoverished pathogen communities whilst different species' ecologies and contrasting life-history traits may lead to different levels of pathogen exposure. Although specific host traits may explain differential particular infections, overall pathogen diversity, richness and prevalence were higher in the truly cosmopolitan, euriphagous and long-distance disperser Eurasian kestrel than in the estenophagous, steppe-specialist, philopatric but long-distance migratory lesser kestrel. Accordingly, the continental population of Eurasian kestrels displayed a higher number (64 vs. 49) as well as more divergent alleles at both MHC class I and class II loci. Detailed analyses of amino acid diversity revealed that significant differences between both species were exclusive to those functionally important codons comprising the antigen binding sites. The lowest pathogen burdens and the smallest but still quite divergent set of MHC alleles (n = 16) were found in island Eurasian kestrels, where the rates of allele fixation at MHC loci seem to have occurred faster than at neutral markers. The results presented in this study would therefore support the role of pathogen diversity and abundance in shaping patterns of genetic variation at evolutionary relevant MHC genes.

Strong philopatry derived from capture-recapture records does not lead to fine-scale genetic differentiation in lesser kestrels.

1. The integration of capture-recapture and molecular approaches can improve our understanding of the consequences of habitat fragmentation on population connectivity. Here we employed microsatellites to test dispersal hypotheses derived from intense and long-term ringing programmes of the lesser kestrel Falco naumanni in Western Europe. 2. Re-encounters of 1308 marked individuals in Spain have revealed that most first-time breeders settled within 10 km from their natal colony, with a negative association between dispersal and geographical distance. Although these findings would predict fine-scale spatial patterns of genetic differentiation, the genetic impact of rarely reported events concerning long-distance effective dispersal (> 100 km) is unknown. 3. First, we investigated a spatially structured and geographically isolated population located in north-eastern Spain, where capture-recapture records and genetic data could be appropriately compared over similar spatial and temporal scales. Spatial autocorrelation analyses (N = 174 nestlings from different broods) did not reveal either significant differences in average relatedness at any distance class nor decreased relatedness as a function of distance. At a broader spatial scale, Bayesian analysis of population structure (N = 432 nestlings) indicated panmixia across Western Europe. However, F(ST) comparisons between four geographically distinct populations indicated low but significant genetic differentiation. 4. Our genetic data would therefore challenge traditional assumptions associating philopatry with the emergence of fine-scale genetic structuring. This could be because even low levels of gene flow are enough to preclude the development of local genetic structure. Nevertheless, the analysis of a geographically isolated and small population from Southern France exemplifies a situation in which restricted dispersal has translated into weak but consistently significant genetic differentiation. 5. Relevant to conservation genetics and evolutionary biology, our results may lessen the genetic concerns derived from population fragmentation at relatively small geographical scales in species with apparently limited dispersal abilities, but raises concerns about increased genetic divergence in small and isolated demes.

Extensive polymorphism and geographical variation at a positively selected MHC class II B gene of the lesser kestrel (Falco naumanni).

Understanding the selective forces that shape genetic variation in natural populations remains a high priority in evolutionary biology. Genes at the major histocompatibility complex (MHC) have become excellent models for the investigation of adaptive variation and natural selection because of their crucial role in fighting off pathogens. Here we present one of the first data sets examining patterns of MHC variation in wild populations of a bird of prey, the lesser kestrel, Falco naumanni. We report extensive polymorphism at the second exon of a putatively functional MHC class II gene, Fana-DAB*1. Overall, 103 alleles were isolated from 121 individuals sampled from Spain to Kazakhstan. Bayesian inference of diversifying selection suggests that several amino acid sites may have experienced strong positive selection (omega = 4.02 per codon). The analysis also suggests a prominent role of recombination in generating and maintaining MHC diversity (rho = 4Nc = 0.389 per codon, theta = 0.017 per codon). Both the Fana-DAB*1 locus and a set of eight polymorphic microsatellite markers revealed an isolation-by-distance pattern across the Western Palaearctic (r = 0.67; P = 0.01 and r = 0.50; P = 0.04, respectively). Nonetheless, geographical variation at the MHC contrasts with relatively uniform distributions in the frequencies of microsatellite alleles. In addition, we found lower fixation rates in the MHC than those predicted by genetic drift after controlling for neutral mitochondrial sequences. Our results therefore underscore the role of balancing selection as well as spatial variations in parasite-mediated selection regimes in shaping MHC diversity when gene flow is limited.

Complex Plumage Patterns Can Be Produced Only with the Contribution of Melanins.

Birds exhibit an extraordinary diversity of plumage pigmentation patterns. It has been overlooked, however, that complex patterns can be produced only with the contribution of melanins because these are the only pigments under direct cellular control. We tested this hypothesis for the first time examining the plumage patterning of all species of extant birds. Thirty-two percent of species show complex plumage patterns, the vast majority (98%) including the contribution of colors produced by melanins. Only 53 species show complex patterns that do not contain melanin-based colors, and these species display unusual colorations and belong to three families where innovative metabolic modifications of conventional carotenoid pigments have been described. While the adaptive functions of complex plumage patterns remain poorly understood and in most cases are ascribed to fulfill camouflage, our findings indicate that such functions will be understood only by considering the synthesis pathway of melanins.

Coloration reflects skin pterin concentration in a red-tailed lizard.

When integumentary tissue pigments are contained in chromatophores, tissue color might not depend exclusively on the amount of pigment. Whether coloration does or does not reflect pigment concentration may be very significant for intraspecific communication, for example when pigment concentration provides fitness-related information. We studied the pigment responsible for the orange/red ventral tail coloring in a lacertid lizard species (Acanthodactylus erythrurus), and whether the color was related to skin pigment concentration. The pigment was identified as a pterin, a higher concentration of which resulted in darker, more red-saturated, redder (less orange) ventral tail skin color. The dorsal tail integument, even though it appears mostly gray to the naked eye, also contained pterins, and furthermore, the dorsal and ventral pterin concentrations were positively correlated. A possible explanation for these results is that pterins accumulate in the skin of the whole tail, even if only needed in the ventral part, but are concealed in the dorsal part. In this way, ventral orange/red coloration would accurately reflect pterin concentration, which provides the basis for a signaling function, while dorsal coloration would become less conspicuous as an anti-predatory mechanism.

Porphyrins produce uniquely ephemeral animal colouration: a possible signal of virginity.

Colours that underlie animal pigmentation can either be permanent or renewable in the short term. Here we describe the discovery of a conspicuous salmon-pink colouration in the base of bustard feathers and down that has never been reported because of its extraordinarily brief expression. HPLC analyses indicated that its constituent pigments are coproporphyrin III and protoporphyrin IX, which are prone to photodegradation. Accordingly, an experimental exposure of feathers of three bustard species to sunlight produced a rapid disappearance of the salmon-pink colouration, together with a marked decrease in reflectance around 670 nm coinciding with the absorption of porphyrin photoproducts. The disappearance of the salmon-pink colouration can occur in a period as short as 12 min, likely making it the most ephemeral colour phenotype in any extant bird. The presence of this colour trait in males performing sexual displays may thus indicate to females a high probability that the males were performing their first displays and would engage in their first copulations in the breeding season. In dominant males, sperm quality decreases over successive copulations, thus porphyrin-based colouration may evolve as a signal of virginity that allows females to maximize their fitness in lek mating systems.