Reduced palatability, fast flight, and tails: decoding the defence arsenal of Eudaminae skipper butterflies in a Neotropical locality.

Prey often rely on multiple defences against predators, such as flight speed, attack deflection from vital body parts, or unpleasant taste, but our understanding on how often and why they are co-exhibited remains limited. Eudaminae skipper butterflies use fast flight and mechanical defences (hindwing tails), but whether they use other defences like unpalatability (consumption deterrence) and how these defences interact have not been assessed. We tested the palatability of 12 abundant Eudaminae species in Peru, using training and feeding experiments with domestic chicks. Further, we approximated the difficulty of capture based on flight speed and quantified it by wing loading. We performed phylogenetic regressions to find any association between multiple defences, body size, and habitat preference. We found a broad range of palatability in Eudaminae, within and among species. Contrary to current understanding, palatability was negatively correlated with wing loading, suggesting that faster butterflies tend to have lower palatability. The relative length of hindwing tails did not explain the level of butterfly palatability, showing that attack deflection and consumption deterrence are not mutually exclusive. Habitat preference (open or forested environments) did not explain the level of palatability either, although butterflies with high wing loading tended to occupy semi-closed or closed habitats. Finally, the level of unpalatability in Eudaminae is size dependent. Larger butterflies are less palatable, perhaps because of higher detectability/preference by predators. Altogether, our findings shed light on the contexts favouring the prevalence of single versus multiple defensive strategies in prey.

Deep divergences among inconspicuously colored clades of Epipedobates poison frogs.

Poison frogs (Dendrobatidae) are famous for their aposematic species, having a combination of diverse color patterns and defensive skin toxins, yet most species in this family are inconspicuously colored and considered non-aposematic. Epipedobates is among the youngest genus-level clades of Dendrobatidae that includes both aposematic and inconspicuous species. Using Sanger-sequenced mitochondrial and nuclear markers, we demonstrate deep genetic divergences among inconspicuous species of Epipedobates but relatively shallow genetic divergences among conspicuous species. Our phylogenetic analysis includes broad geographic sampling of the inconspicuous lineages typically identified as E. boulengeri and E. espinosai, which reveals two putative new species, one in west-central Colombia (E. sp. 1) and the other in north-central Ecuador (E. aff. espinosai). We conclude that E. darwinwallacei is a junior subjective synonym of E. espinosai. We also clarify the geographic distributions of inconspicuous Epipedobates species including the widespread E. boulengeri. We provide a qualitative assessment of the phenotypic diversity in each nominal species, with a focus on the color and pattern of inconspicuous species. We conclude that Epipedobates contains eight known valid species, six of which are inconspicuous. A relaxed molecular clock analysis suggests that the most recent common ancestor of Epipedobates is âˆ¼11.1 million years old, which nearly doubles previous estimates. Last, genetic information points to a center of species diversity in the Chocó at the southwestern border of Colombia with Ecuador. A Spanish translation of this text is available in the supplementary materials.

Outcomes of multifarious selection on the evolution of visual signals.

Multifarious sources of selection shape visual signals and can produce phenotypic divergence. Theory predicts that variance in warning signals should be minimal due to purifying selection, yet polymorphism is abundant. While in some instances divergent signals can evolve into discrete morphs, continuously variable phenotypes are also encountered in natural populations. Notwithstanding, we currently have an incomplete understanding of how combinations of selection shape fitness landscapes, particularly those which produce polymorphism. We modelled how combinations of natural and sexual selection act on aposematic traits within a single population to gain insights into what combinations of selection favours the evolution and maintenance of phenotypic variation. With a rich foundation of studies on selection and phenotypic divergence, we reference the poison frog genus Oophaga to model signal evolution. Multifarious selection on aposematic traits created the topology of our model's fitness landscape by approximating different scenarios found in natural populations. Combined, the model produced all types of phenotypic variation found in frog populations, namely monomorphism, continuous variation and discrete polymorphism. Our results afford advances into how multifarious selection shapes phenotypic divergence, which, along with additional modelling enhancements, will allow us to further our understanding of visual signal evolution.

Thin-fingered fiddler crabs display a natural preference for UV light cues but show no sensory bias to other hypertrophied claw coloration.

Although color cues play an important role in sociosexual signaling, when communication is exploited as a foraging cue by predators, prey must pay the due cost for sustaining information interchange. In this regard, fiddler crabs draw attention by having flashy enlarged claws that could potentially attract the interest of many predators. Surprisingly, the adaptive function of claw coloration in fiddler crabs is not entirely understood and have barely been studied in American species. Here, we examine the spectral reflectance of thin-fingered (Leptuca leptodactyla) fiddler crabs' hypertrophied claws and evaluate whether female conspecifics prefer UV-light or other color cues. We test whether male claws reflect UV light and whether females use UV light and/or other color cues when making mating decisions. Our results reveal that only the most flamboyant enlarged claws should be detected by female conspecifics, which show a clear preference for male UV signals. Moreover, while the anterior portion of the enlarged claws (pointing towards conspecifics) reflect UV light, claws' dorsal portion and carapaces (which possibly point upwards and might attract airborne predators) do not show an UV peak. We also discuss the possibility of hypertrophied claws working as honest aposematic signals or playing an important role as decoys.

Mimicry can drive convergence in structural and light transmission features of transparent wings in Lepidoptera.

Müllerian mimicry is a positive interspecific interaction, whereby co-occurring defended prey species share a common aposematic signal. In Lepidoptera, aposematic species typically harbour conspicuous opaque wing colour patterns with convergent optical properties among co-mimetic species. Surprisingly, some aposematic mimetic species have partially transparent wings, raising the questions of whether optical properties of transparent patches are also convergent, and of how transparency is achieved. Here, we conducted a comparative study of wing optics, micro and nanostructures in neotropical mimetic clearwing Lepidoptera, using spectrophotometry and microscopy imaging. We show that transparency, as perceived by predators, is convergent among co-mimics in some mimicry rings. Underlying micro- and nanostructures are also sometimes convergent despite a large structural diversity. We reveal that while transparency is primarily produced by microstructure modifications, nanostructures largely influence light transmission, potentially enabling additional fine-tuning in transmission properties. This study shows that transparency might not only enable camouflage but can also be part of aposematic signals.

Initial Phylotranscriptomic Confirmation of Homoplastic Evolution of the Conspicuous Coloration and Bufoniform Morphology of Pumpkin-Toadlets in the Genus Brachycephalus.

The genus Brachycephalus is a fascinating group of miniaturized anurans from the Brazilian Atlantic Forest, comprising the conspicuous, brightly colored pumpkin-toadlets and the cryptic flea-toads. Pumpkin-toadlets are known to contain tetrodotoxins and therefore, their bright colors may perform an aposematic function. Previous studies based on a limited number of mitochondrial and nuclear-encoded markers supported the existence of two clades containing species of pumpkin-toadlet phenotype, but deep nodes remained largely unresolved or conflicting between data sets. We use new RNAseq data of 17 individuals from nine Brachycephalus species to infer their evolutionary relationships from a phylogenomic perspective. Analyses of almost 5300 nuclear-encoded ortholog protein-coding genes and full mitochondrial genomes confirmed the existence of two separate pumpkin-toadlet clades, suggesting the convergent evolution (or multiple reversals) of the bufoniform morphology, conspicuous coloration, and probably toxicity. In addition, the study of the mitochondrial gene order revealed that three species (B. hermogenesi, B. pitanga, and B. rotenbergae) display translocations of different tRNAs (NCY and CYA) from the WANCY tRNA cluster to a position between the genes ATP6 and COIII, showing a new mitochondrial gene order arrangement for vertebrates. The newly clarified phylogeny suggests that Brachycephalus has the potential to become a promising model taxon to understand the evolution of coloration, body plan and toxicity. Given that toxicity information is available for only few species of Brachycephalus, without data for any flea-toad species, we also emphasize the need for a wider screening of toxicity across species, together with more in-depth functional and ecological study of their phenotypes.

A new arc-striped species of Corydoras Lacépède, 1803 (Teleostei: Callichthyidae) from the Peruvian Amazon.

A new Corydoras is described from the Blanco and Ucayali river basins in Peru. The new species can be distinguished from its congeners by having the following features: (I) posterior margin of dorsal-fin spine with laminar serrations directed towards the origin of the spine; (II) a long, wide, arched, and continuous black stripe, which runs parallel to the dorsal profile of the body, extending at least from the region below anterior origin of dorsal fin to the anterior half of the ventral caudal-fin lobe; (III) a black stripe transversally crossing the eye, forming the typical mask-like blotch; mask clearly not fused to arched stripe in most specimens; some specimens with mask separated from arched stripe by a thin line around the suture between neurocranium (in the region composed by the posteroventral margin of parieto-supraoccipital plus the posterodorsal margin of the compound pterotic) and first dorsolateral body plate; (IV) posterior margin of pectoral-fin spine with laminar serrations directed towards the origin of the spine; (V) pointed snout, presenting a long mesethmoid, with anterior tip larger than 50% of the entire length of the bone; and (V) ventral surface of trunk covered by small, non-coalescent platelets. A discussion on the possible positive adaptive value of the arc-striped color pattern is also provided.

Toxicity and Alkaloid Profiling of the Skin of the Golfo Dulcean Poison Frog Phyllobates vittatus (Dendrobatidae).

Frogs in the genus Phyllobates are known for the presence of batrachotoxin, a highly toxic alkaloid, in their skin. Nevertheless, Phyllobates frogs from Costa Rica and Panama (P. lugubris and P. vittatus) are considered non-toxic, as they have been reported to harbor low concentrations of this alkaloid. However, the potential toxicity of Central American Phyllobates has not been assessed experimentally. Our goal was to determine the toxicity of the whole skin of P. vittatus, an endemic species from the Southeastern Pacific region of Costa Rica. We performed median lethal dose (LD50) tests in mice to determine general toxicity, and an irritant assay based on the behavioral responses of mice to subcutaneous injection, to determine differences in irritability, as a measure of toxicity, among three study localities. Using UPLC-ESI-QTOF, we obtained chemical profiles of the methanolic extract of frog skins. Due to the absence of mortality at the studied doses, we were unable to estimate LD50. However, we recorded a list of toxicity symptoms in mice that are consistent with cardiotoxic effects, and found that mice presented more symptoms at higher concentrations of skin extracts during the first hour of the LD50 assays, recovering completely at all doses by the end of the assay. On the other hand, we did not detect differences in irritability among studied localities. Additionally, we putatively identified three toxic alkaloids (Batrachotoxinin A, DHQ 251A and Lehm 275A). This study provides the first experimental data on the toxicity and associated symptoms in mice, as well as the chemical profile of the skin of P. vittatus. We suggest that the skin alkaloids of P. vitattus may confer a chemical defense towards predators.

Prey and predators perceive orb-web spider conspicuousness differently: evaluating alternative hypotheses for color polymorphism evolution.

Color polymorphisms have been traditionally attributed to apostatic selection. The perception of color depends on the visual system of the observer. Theoretical models predict that differently perceived degrees of conspicuousness by two predator and prey species may cause the evolution of polymorphisms in the presence of anti-apostatic and apostatic selection. The spider Gasteracantha cancriformis (Araneidae) possesses several conspicuous color morphs. In orb-web spiders, the prey attraction hypothesis states that conspicuous colors are prey lures that increase spider foraging success via flower mimicry. Therefore, polymorphism could be maintained if each morph attracted a different prey species (multiple prey hypothesis) and each spider mimicked a different flower color (flower mimicry hypothesis). Conspicuous colors could be a warning signal to predators because of the spider's hard abdomen and spines. Multiple predators could perceive morphs differently and exert different degrees of selective pressures (multiple predator hypothesis). We explored these 3 hypotheses using reflectance data and color vision modeling to estimate the chromatic and achromatic contrast of G. cancriformis morphs as perceived by several potential prey and predator taxa. Our results revealed that individual taxa perceive the conspicuousness of morphs differently. Therefore, the multiple prey hypothesis and, in part, the multiple predator hypothesis may explain the evolution of color polymorphism in G. cancriformis, even in the presence of anti-apostatic selection. The flower mimicry hypothesis received support by color metrics, but not by color vision models. Other parameters not evaluated by color vision models could also affect the perception of morphs and influence morph survival and polymorphism stability.

Widespread Occurrence of Black-Orange-Black Color Pattern in Hymenoptera.

Certain color patterns in insects show convergent evolution reflecting potentially important biological functions, for example, aposematism and mimicry. This phenomenon has been most frequently documented in Lepidoptera and Coleoptera, but has been less well investigated in Hymenoptera. It has long been recognized that many hymenopterans, especially scelionids (Platygastridae), show a recurring pattern of black head, orange/red mesosoma, and black metasoma (BOB coloration). However, the taxonomic distribution of this striking color pattern has never been documented across the entire order. The main objective of our research was to provide a preliminary tabulation of this color pattern in Hymenoptera, through examination of museum specimens and relevant literature. We included 11 variations of the typical BOB color pattern but did not include all possible variations. These color patterns were found in species belonging to 23 families of Hymenoptera, and was most frequently observed in scelionids, evaniids, and mutillids, but was relatively infrequent in Cynipoids, Diaprioids, Chalcidoids, and Apoids. The widespread occurrence of this color pattern in Hymenoptera strongly suggests convergent evolution and a potentially important function. The BOB color pattern was found in species from all biogeographic regions and within a species it was usually present in both sexes (with a few notable exceptions). In better studied tropical regions, such as Costa Rica, this color pattern was more common in species occurring at lower elevations (below 2,000 m). The biology of the tabulated taxa encompasses both ecto- and endoparasitoids, idiobionts and koinobionts, from a diversity of hosts, as well as phytophagous sawflies.

Toxicity of the feathers of Yellow Grosbeak, Pheucticus chrysopeplus (Passeriformes: Cardinalidae), a chemically defended neotropical bird / Toxicidad de las plumas del Picogrueso amarillo, Pheucticus chrysopeplus (Passeriformes: Cardinalidae), un ave neotropical con defensa química

Abstract Chemical defense is a widespread mechanism on many animals and plants. However, just a few cases are known for avian species. In this study we evaluate the toxicity of Pheucticus chrysopeplus feather extract via lethality test with brine shrimp (Artemia salina) as an in vivo model. Mortality of A. salina was evaluated after 24 hour exposure to artificial seawater, methanol, and the methanolic feather extract. Kruskal-Wallis test showed a significant difference in mortality between treatments (X2 = 65.25, P < 0.0001, n = 50). With this we describe P. chrysopeplus as the first known toxic avian species of Guatemala and Central America, raising awareness about its conservation and the identification of the toxic substance present in its feathers. We also highlight the possible mimicry mechanism taking part between P. chrysopeplus and two sympatric oriole species (Icterus pectoralis and I. pustulatus).(AU)

Resumen La defensa química es un mecanismo que se encuentra presente en varios animales y plantas. Sin embargo, pocos casos son conocidos para especies de aves. En este estudio evaluamos la toxicidad de extractos de plumas de Pheucticus chrysopeplus con un ensayo de letalidad utilizando artemia (Artemia salina) como modelo in vivo. La mortalidad de A. salina se evaluó luego de ser expuesta por 24 horas a agua marina artificial, metanol y extracto metanólico de plumas de P. chrysopeplus. La prueba de Kruskal-Wallis mostró que existe una diferencia significativa entre los porcentajes de mortalidad de los tratamientos evaluados (X2 = 65.25, P < 0.0001, n = 50). Con esto, describimos a P. chrysopeplus como la primera especie de ave tóxica reportada para Guatemala y Centroamérica, resaltando la importancia de su conservación, así como la identificación de la sustancia tóxica presente en sus plumas. También destacamos el posible mecanismo de mimetismo que podría estar ocurriendo entre P. Chrysopeplus y dos especies simpátricas de orioles (Icterus pectoralis e I. pustulatus).(AU)

Radiation of the polymorphic Little Devil poison frog (Oophaga sylvatica) in Ecuador.

Some South American poison frogs (Dendrobatidae) are chemically defended and use bright aposematic colors to warn potential predators of their unpalatability. Aposematic signals are often frequency-dependent where individuals deviating from a local model are at a higher risk of predation. However, extreme diversity in the aposematic signal has been documented in poison frogs, especially in Oophaga. Here, we explore the phylogeographic pattern among color-divergent populations of the Little Devil poison frog Oophaga sylvatica by analyzing population structure and genetic differentiation to evaluate which processes could account for color diversity within and among populations. With a combination of PCR amplicons (three mitochondrial and three nuclear markers) and genome-wide markers from a double-digested RAD (ddRAD) approach, we characterized the phylogenetic and genetic structure of 199 individuals from 13 populations (12 monomorphic and 1 polymorphic) across the O. sylvatica distribution. Individuals segregated into two main lineages by their northern or southern latitudinal distribution. A high level of genetic and phenotypic polymorphism within the northern lineage suggests ongoing gene flow. In contrast, low levels of genetic differentiation were detected among the southern lineage populations and support recent range expansions from populations in the northern lineage. We propose that a combination of climatic gradients and structured landscapes might be promoting gene flow and phylogenetic diversification. Alternatively, we cannot rule out that the observed phenotypic and genomic variations are the result of genetic drift on near or neutral alleles in a small number of genes.

Both Palatable and Unpalatable Butterflies Use Bright Colors to Signal Difficulty of Capture to Predators.

Birds are able to recognize and learn to avoid attacking unpalatable, chemically defended butterflies after unpleasant experiences with them. It has also been suggested that birds learn to avoid prey that are efficient at escaping. This, however, remains poorly documented. Here, we argue that butterflies may utilize a variety of escape tactics against insectivorous birds and review evidence that birds avoid attacking butterflies that are hard to catch. We suggest that signaling difficulty of capture to predators is a widespread phenomenon in butterflies, and this ability may not be limited to palatable butterflies. The possibility that both palatable and unpalatable species signal difficulty of capture has not been fully explored, but helps explain the existence of aposematic coloration and escape mimicry in butterflies lacking defensive chemicals. This possibility may also change the role that putative Müllerian and Batesian mimics play in a variety of classical mimicry rings, thus opening new perspectives in the evolution of mimicry in butterflies.

Warning signals are seductive: relative contributions of color and pattern to predator avoidance and mate attraction in Heliconius butterflies.

Visual signaling in animals can serve many uses, including predator deterrence and mate attraction. In many cases, signals used to advertise unprofitability to predators are also used for intraspecific communication. Although aposematism and mate choice are significant forces driving the evolution of many animal phenotypes, the interplay between relevant visual signals remains little explored. Here, we address this question in the aposematic passion-vine butterfly Heliconius erato by using color- and pattern-manipulated models to test the contributions of different visual features to both mate choice and warning coloration. We found that the relative effectiveness of a model at escaping predation was correlated with its effectiveness at inducing mating behavior, and in both cases wing color was more predictive of presumptive fitness benefits than wing pattern. Overall, however, a combination of the natural (local) color and pattern was most successful for both predator deterrence and mate attraction. By exploring the relative contributions of color versus pattern composition in predation and mate preference studies, we have shown how both natural and sexual selection may work in parallel to drive the evolution of specific animal color patterns.

Coarse dark patterning functionally constrains adaptive shifts from aposematism to crypsis in strawberry poison frogs.

Ecological specialization often requires tight coevolution of several traits, which may constrain future evolutionary pathways and make species more prone to extinction. Aposematism and crypsis represent two specialized adaptations to avoid predation. We tested whether the combined effects of color and pattern on prey conspicuousness functionally constrain or facilitate shifts between these two adaptations. We combined data from 17 natural populations of strawberry poison frogs, Oophaga pumilio with an experimental approach using digitalized images of frogs and chickens as predators. We show that bright coloration often co-occurs with coarse patterning among the natural populations. Dull green frogs with coarse patterning are rare in nature but in the experiment they were as easily detected as bright red frogs suggesting that this trait combination represents a transient evolutionary state toward aposematism. Hence, a gain of either bright color or coarse patterning leads to conspicuousness, but a transition back to crypsis would be functionally constrained in populations with both bright color and coarse patterning by requiring simultaneous changes in two traits. Thus, populations (or species) signaling aposematism by conspicuous color should be less likely to face an evolutionary dead end and more likely to radiate than populations with both conspicuous color and coarse patterning.

Phenotypic and molecular variation in the green and black poison-dart frog Dendrobates auratus (Anura: Dendrobatidae) from Costa Rica / Variación fenotípica y molecular en la rana dardo venenoso verde y negra Dendrobates auratus (Anura: Dendrobatidae) de Costa Rica

The green and black poison-dart frog Dendrobates auratus exhibits high intraspecific variation in hue color and pattern throughout its range, making it a very popular species in the pet trade. We analyzed the correspondence between color variation and molecular variation of D. auratus from Costa Rica using RAPD analysis. Twenty-six random primers were analyzed for variation in 99 individuals from seven populations. Color pattern was scored from digital images of the dorsal and ventral views. In general, frogs from the Caribbean coast had significantly more light coloration than black color but cannot be grouped by population based only on hue pattern. Only 3 RAPD primers were found to be polymorphic, representing a total of 16 loci. Most of the molecular variation encountered here occurs within populations, thus making unclear the degree of population structure and differentiation. Further examination of COI mtDNA sequences from our samples also supports these results. Partial Mantel correlations suggested that the pattern of molecular variation is not congruent with the variation in color pattern in this species, an outcome that is discussed in terms of phenotypic evolution. Rev. Biol. Trop. 57 (Suppl. 1): 313-321. Epub 2009 November 30.

La rana venenosa Dendrobates auratus posee una gran variación intraespecífica en tonos y patrones de coloración a lo largo de toda su distribución, lo que la hace una especie muy reconocible entre las especies de dendrobátidos. Analizamos la correspondencia entre variación de coloración y variación molecular de D. auratus de Costa Rica empleando análisis de RAPDs. La variación resultante en veintiséis "primers" aleatorios fue analizada en 93 individuos de siete localidades en Costa Rica. El patrón de coloración fue evaluado de imágenes digitales del dorso y vientre para los mismos individuos. En general, las ranas provenientes de localidades en la costa Caribe tienen significativamente una coloración más clara, con menos proporción de color negro que las de localidades en la vertiente Pacífica, pero no pueden ser agrupadas por localidad basadas simplemente en el patrón de coloración. Solamente tres RAPD "primers" fueron encontrados polimórficos, representando un total de 16 loci. Mucha de la variación molecular encontrada habita dentro de poblaciones, lo que hace difícil determinar el grado de estructura poblacional y diferenciación. La reexaminación posterior de secuencias del gen mitocondrial CO1 también apoya estos resultados. Correlaciones parciales de matrices (test de Mantel) sugieren que el patrón de variación molecular no es congruente con la variación en el patrón de coloración en esta especie, un resultado que es discutido en términos de evolución fenotípica.

DIFFERENTIAL AVOIDANCE OF CORAL SNAKE BANDED PATTERNS BY FREE-RANGING AVIAN PREDATORS IN COSTA RICA.

Empirical studies of mimicry have rarely been conducted under natural conditions. Field investigations of some lepidopteran systems have provided a bridge between experiments examining artificial situations and the mimicry process in nature, but these systems do not include all types of mimicry. The presence of dangerous or deadly models is thought to alter the usual rules for mimicry complexes. In particular, a deadly model is expected to protect a wide variety of mimics. Avoidance of different types of mimics should vary according to how closely they resemble the model. Coral snake mimicry complexes in the neotropics may provide natural systems in which these ideas can be examined, but there is no direct evidence that the patterns of venomous coral snakes or potential mimics are avoided in the wild. Plasticine replicas of snakes were used to assess the frequency of avian predation attempts as a function of color pattern. Avian predators left identifiable marks on the replicas, the position of which indicated that replicas were perceived as potentially dangerous prey items by birds. The number of attacks on unmarked brown replicas was greater than that on tricolor coral snake banded replicas. This result was true whether replicas were placed on natural or plain white backgrounds, suggesting that coral snake banded patterns function aposematically. In a separate experiment, replicas representing all six patterns of proposed coral mimics at the study site were attacked less often than unmarked brown replicas. Within these six banded patterns, some were attacked significantly more often than others. This study provides direct field evidence that coral snake banded patterns are avoided by free-ranging avian predators and supports theoretical predictions about mimicry systems involving deadly models.