Rapid and reversible humidity-dependent colour change by water film formation in a scaled springtail.

Colour is often not a static trait but can change over time either through biotic or abiotic factors. Humidity-dependent colour change can occur through either morphological change (e.g. to feather barbules in birds) or by the replacement of air by water causing a shift in refractive index, as seen in arthropod multi-layer cuticles or scales. The scaled springtail Lepidocyrtus cyaneus has scales that produce colour largely via thin film interference from their lamina. We observed a marked colour change from golden to violet/purple coloration in humid conditions. Light microscopy, micro-spectrophotometry, contact angle goniometry and optical modelling indicate that the formation of a thin film of water on top of the hydrophilic scales increases their laminar thin film thickness, causing a shift towards violet/purple colour. Evaporation of the water film causes the metallic golden colour to return. This constitutes a remarkably rapid colour change (in the order of seconds), only limited by the speed of water film condensation and evaporation, that may serve as inspiration for new dynamically coloured materials and sensors.

Influence of Core Type and Shell Thickness on Avian-Inspired Structural Colors Produced from Melanin Nanoparticle Assemblies.

Hollow melanosomes found in iridescent bird feathers, including violet-backed starlings and wild turkeys, enable the generation of diverse structural colors. It has been postulated that the high refractive index (RI) contrast between melanin (1.74) and air (1.0) results in brighter and more saturated colors. This has led to several studies that have synthesized hollow synthetic melanin nanoparticles and fabricated colloidal nanostructures to produce synthetic structural colors. However, these studies use hollow nanoparticles with thin shells (<20 nm), even though shell thicknesses as high as 100 nm have been observed in natural melanosomes. Here, we combine experimental and computational approaches to examine the influence of the varying polydopamine (PDA, synthetic melanin) shell thickness (0-100 nm) and core material on structural colors. Experimentally, a concomitant change in overall particle size and RI contrast makes it difficult to interpret the effect of a hollow or solid core on color. Thus, we utilize finite-difference time-domain (FDTD) simulations to uncover the effect of shell thickness and core on structural colors. Our FDTD results highlight that hollow particles with thin shells have substantially higher saturation than same-sized solid and core-shell particles. These results would benefit a wide range of applications including paints, coatings, and cosmetics.

Mechanism of structural colors in binary mixtures of nanoparticle-based supraballs.

Inspired by structural colors in avian species, various synthetic strategies have been developed to produce noniridescent, saturated colors using nanoparticle assemblies. Nanoparticle mixtures varying in particle chemistry and size have additional emergent properties that affect the color produced. For complex multicomponent systems, understanding the assembled structure and a robust optical modeling tool can empower scientists to identify structure-color relationships and fabricate designer materials with tailored color. Here, we demonstrate how we can reconstruct the assembled structure from small-angle scattering measurements using the computational reverse-engineering analysis for scattering experiments method and use the reconstructed structure in finite-difference time-domain calculations to predict color. We successfully, quantitatively predict experimentally observed color in mixtures containing strongly absorbing nanoparticles and demonstrate the influence of a single layer of segregated nanoparticles on color produced. The versatile computational approach that we present is useful for engineering synthetic materials with desired colors without laborious trial-and-error experiments.

Hydrophobic Melanin via Post-Synthetic Modification for Controlled Self-Assembly.

Allomelanin is a class of nitrogen-free melanin mostly found in fungi and, like all naturally occurring melanins, is hydrophilic. Herein, we develop a facile method to modify synthetic hydrophilic allomelanin to yield hydrophobic derivatives through post-synthetic modifications. Amine-functionalized molecules of various kinds can be conjugated to allomelanin nanoparticles under mild conditions with high loading efficiencies. Hydrophobicity is conferred by introducing amine-terminated alkyl groups with different chain lengths. We demonstrate that the resulting hydrophobic allomelanin nanoparticles undergo air/water interfacial self-assembly in a controlled fashion, which enables the generation of large-scale and uniform structural colors. This work provides an efficient and tunable surface chemistry modification strategy to broaden the scope of synthetic melanin structure and function beyond the known diversity found in nature.

A combination of red structural and pigmentary coloration in the eyespot of a copepod.

While the specific mechanisms of colour production in biological systems are diverse, the mechanics of colour production are straightforward and universal. Colour is produced through the selective absorption of light by pigments, the scattering of light by nanostructures or a combination of both. When Tigriopus californicus copepods were fed a carotenoid-limited diet of yeast, their orange-red body coloration became faint, but their eyespots remained unexpectedly bright red. Raman spectroscopy indicated a clear signature of the red carotenoid pigment astaxanthin in eyespots; however, refractive index matching experiments showed that eyespot colour disappeared when placed in ethyl cinnamate, suggesting a structural origin for the red coloration. We used transmission electron microscopy to identify consecutive nanolayers of spherical air pockets that, when modelled as a single thin film layer, possess the correct periodicity to coherently scatter red light. We then performed microspectrophotometry to quantify eyespot coloration and confirmed a distinct colour difference between the eyespot and the body. The observed spectral reflectance from the eyespot matched the reflectance predicted from our models when considering the additional absorption by astaxanthin. Together, this evidence suggests the persistence of red eyespots in copepods is the result of a combination of structural and pigmentary coloration.

Springtail coloration at a finer scale: mechanisms behind vibrant collembolan metallic colours.

The mechanisms and evolution of metallic structural colours are of both fundamental and applied interest, yet most work in arthropods has focused on derived butterflies and beetles with distinct hues. In particular, basal hexapods-groups with many scaled, metallic representatives-are currently poorly studied and controversial, with some recent studies suggesting either that thin-film (lamina thickness) or diffraction grating (longitudinal ridges, cross-ribs) elements produce these colours in early Lepidoptera and one springtail (Collembola) species. Especially the collembolan basal scale design, consisting of a single lamina and longitudinal ridges with smooth valleys lacking cross-ribs, makes them an interesting group to explore the mechanisms of metallic coloration. Using microspectroscopy, Raman spectroscopy, electron microscopy and finite-difference time-domain optical modelling, we investigated scale colour in seven springtail species that show clear metallic coloration. Reflectance spectra are largely uniform and exhibit a broadband metallic/golden coloration with peaks in the violet/blue region. Our simulations confirm the role of the longitudinal ridges, working in conjunction with thin-film effects to produce a broadband metallic coloration. Broadband coloration occurs through spatial colour mixing, which probably results from nanoscale variation in scale thickness and ridge height and distance. These results provide crucial insights into the colour production mechanisms in a basal scale design and highlight the need for further investigation of scaled, basal arthropods.

Evolution of eggshell structure in relation to nesting ecology in non-avian reptiles.

Amniotic eggs are multifunctional structures that enabled early tetrapods to colonize the land millions of years ago, and are now the reproductive mode of over 70% of all terrestrial amniotes. Eggshell morphology is at the core of animal survival, mediating the interactions between embryos and their environment, and has evolved into a massive diversity of forms and functions in modern reptiles. These functions are critical to embryonic survival and may serve as models for new antimicrobial and/or breathable membranes. However, we still lack critical data on the basic structural and functional properties of eggs, particularly of reptiles. Here, we first characterized egg shape, shell thickness, porosity, and mineralization of eggs from 91 reptile species using optical images, scanning electron microscopy, and micro computed tomography, and collected data on nesting ecology from the literature. We then used comparative analyses to test hypotheses on the selective pressures driving their evolution. We hypothesized that eggshell morphology has evolved to protect shells from physical damage and desiccation, and, in support, found a positive relationship between thickness and precipitation, and a negative relationship between porosity and temperature. Although mineralization varied extensively, it was not correlated with nesting ecology variables. Ancestral state reconstructions show thinning and increased porosity over evolutionary time in squamates, but the opposite in turtles and crocodilians. Egg shape, size, porosity and calcification were correlated, suggesting potential structural or developmental tradeoffs. This study provides new data and insights into the morphology and evolution of reptile eggs, and raises numerous questions for additional research.

Thermoregulation and heat exchange in ospreys (Pandion haliaetus).

The osprey (Pandion haliaetus) is a cosmopolitan and long-distant migrant, found at all thermal extremes ranging from polar to tropical climates. Since ospreys may have an unusually flexible thermal physiology due to their migration over, and use of, a wide range of habitats, they represent an interesting study system to explore thermoregulatory adaptations in a raptor. In this study, we investigated the efficiency of heat exchange between body and environment in ospreys using micro-computed tomography (µ-CT), infrared thermography and behavioral observations. µ-CT revealed that the osprey bill has its largest potential for heat exchange at the proximal bill region, where arteries are situated most closely under the surface. However, thermal images of 10 juvenile ospreys showed that the bill contributes to only 0.3% of the bird's total heat exchange. The long legs and protruding claws played a more prominent role as heat dissipation areas with a contribution of 6% and 7%, respectively. Operative thresholds, i.e. the ambient temperature below which heat is lost, were high (>38.5 °C) in these body parts. However, we found no indication of active regulation of heat exchange. Instead we observed multiple behavioral adaptations starting at relatively low ambient temperatures. At 26.3 °C ospreys had a 50% probability of showing panting behavior and above 27.9 °C they additionally spread their wings to enable heat dissipation from the less insulated ventral side. The thermal images revealed that at an ambient temperature of 32.1 °C ospreys had a 50% probability of developing a ≥2 °C and up to 7.5 °C colder stripe on the head, which was likely caused by cutaneous evaporation. Our observations suggest that ospreys more strongly rely on behavioral mechanisms than on active thermal windows to cope with heat stress. This study not only improves our understanding of the role of different body parts in ospreys' total heat exchange with the environment but further provides an insight about additional adaptations of this raptor to cope with heat stress.

Substrate thermal properties influence ventral brightness evolution in ectotherms.

The thermal environment can affect the evolution of morpho-behavioral adaptations of ectotherms. Heat is transferred from substrates to organisms by conduction and reflected radiation. Because brightness influences the degree of heat absorption, substrates could affect the evolution of integumentary optical properties. Here, we show that vipers (Squamata:Viperidae) inhabiting hot, highly radiative and superficially conductive substrates have evolved bright ventra for efficient heat transfer. We analyzed the brightness of 4161 publicly available images from 126 species, and we found that substrate type, alongside latitude and body mass, strongly influences ventral brightness. Substrate type also significantly affects dorsal brightness, but this is associated with different selective forces: activity-pattern and altitude. Ancestral estimation analysis suggests that the ancestral ventral condition was likely moderately bright and, following divergence events, some species convergently increased their brightness. Vipers diversified during the Miocene and the enhancement of ventral brightness may have facilitated the exploitation of arid grounds. We provide evidence that integument brightness can impact the behavioral ecology of ectotherms.

The golden age of arthropods: ancient mechanisms of colour production in body scales.

Insect colour is extremely diverse and produced by a large number of pigmentary and nanostructural mechanisms. Considerable research has been dedicated to these optical mechanisms, with most of it focused on chromatic colours, such as blues and greens, and less on achromatic colours like white and gold. Moreover, studies on the evolution of these colours are less common and largely limited to inferences from extant organisms, in part because of the limited amount and types of available fossil material. Here, we directly compare nanostructure and colour of extant and amber-preserved (approx. 15 and 99 Myr old, respectively) gold-coloured representatives of micromoths (Lepidoptera: Micropterigidae) and springtails (Collembola: Tomoceridae). Using electron microscopy, microspectrophotometry and finite domain time difference optical modelling, we show that golden coloration in the extant micromoth is produced by nanometre-scale crossribs that function as zero-order diffraction gratings and in the springtail by a diffraction grating without crossribs. Surprisingly, nanostructure and thus predicted colour of the amber-preserved specimens were nearly identical to those of their extant counterparts. Removal of amber enabled direct colour measurement of the fossil micromoth and further revealed that its colour matched both that of the extant specimen and the predicted colour, providing further support for our optical models. Our data thus clearly show an early origin and striking conservation of scale nanostructures and golden coloration, suggesting strong selection pressure either on the colour itself or on the mechanisms that produce the colour. Furthermore, we show the thus-far untapped potential for the use of amber-preserved specimens in studies on the evolution of organismal coloration.

Evidence for Faster X Chromosome Evolution in Spiders.

In species with chromosomal sex determination, X chromosomes are predicted to evolve faster than autosomes because of positive selection on recessive alleles or weak purifying selection. We investigated X chromosome evolution in Stegodyphus spiders that differ in mating system, sex ratio, and population dynamics. We assigned scaffolds to X chromosomes and autosomes using a novel method based on flow cytometry of sperm cells and reduced representation sequencing. We estimated coding substitution patterns (dN/dS) in a subsocial outcrossing species (S. africanus) and its social inbreeding and female-biased sister species (S. mimosarum), and found evidence for faster-X evolution in both species. X chromosome-to-autosome diversity (piX/piA) ratios were estimated in multiple populations. The average piX/piA estimates of S. africanus (0.57 [95% CI: 0.55-0.60]) was lower than the neutral expectation of 0.75, consistent with more hitchhiking events on X-linked loci and/or a lower X chromosome mutation rate, and we provide evidence in support of both. The social species S. mimosarum has a significantly higher piX/piA ratio (0.72 [95% CI: 0.65-0.79]) in agreement with its female-biased sex ratio. Stegodyphus mimosarum also have different piX/piA estimates among populations, which we interpret as evidence for recurrent founder events. Simulations show that recurrent founder events are expected to decrease the piX/piA estimates in S. mimosarum, thus underestimating the true effect of female-biased sex ratios. Finally, we found lower synonymous divergence on X chromosomes in both species, and the male-to-female substitution ratio to be higher than 1, indicating a higher mutation rate in males.

Male spiders control offspring sex ratio through greater production of female-determining sperm.

Sex allocation theory predicts that when sons and daughters have different reproductive values, parents should adjust offspring sex ratio towards the sex with the higher fitness return. Haplo-diploid species directly control offspring sex ratio, but species with chromosomal sex determination (CSD) were presumed to be constrained by Mendelian segregation. There is now increasing evidence that CSD species can adjust sex ratio strategically, but the underlying mechanism is not well understood. One hypothesis states that adaptive control is more likely to evolve in the heterogametic sex through a bias in gamete production. We investigated this hypothesis in males as the heterogametic sex in two social spider species that consistently show adaptive female-biased sex ratio and in one subsocial species that is characterized by equal sex ratio. We quantified the production of male (0) and female (X) determining sperm cells using flow cytometry, and show that males of social species produce significantly more X-carrying sperm than 0-sperm, on average 70%. This is consistent with the production of more daughters. Males of the subsocial species produced a significantly lower bias of 54% X-carrying sperm. We also investigated whether inter-genomic conflict between hosts and their endosymbionts may explain female bias. Next generation sequencing showed that five common genera of bacterial endosymbionts known to affect sex ratio are largely absent, ruling out that endosymbiont bacteria bias sex ratio in social spiders. Our study provides evidence for paternal control over sex allocation through biased gamete production as a mechanism by which the heterogametic sex in CSD species adaptively adjust offspring sex ratio.

The house spider genome reveals an ancient whole-genome duplication during arachnid evolution.

BACKGROUND: The duplication of genes can occur through various mechanisms and is thought to make a major contribution to the evolutionary diversification of organisms. There is increasing evidence for a large-scale duplication of genes in some chelicerate lineages including two rounds of whole genome duplication (WGD) in horseshoe crabs. To investigate this further, we sequenced and analyzed the genome of the common house spider Parasteatoda tepidariorum. RESULTS: We found pervasive duplication of both coding and non-coding genes in this spider, including two clusters of Hox genes. Analysis of synteny conservation across the P. tepidariorum genome suggests that there has been an ancient WGD in spiders. Comparison with the genomes of other chelicerates, including that of the newly sequenced bark scorpion Centruroides sculpturatus, suggests that this event occurred in the common ancestor of spiders and scorpions, and is probably independent of the WGDs in horseshoe crabs. Furthermore, characterization of the sequence and expression of the Hox paralogs in P. tepidariorum suggests that many have been subject to neo-functionalization and/or sub-functionalization since their duplication. CONCLUSIONS: Our results reveal that spiders and scorpions are likely the descendants of a polyploid ancestor that lived more than 450 MYA. Given the extensive morphological diversity and ecological adaptations found among these animals, rivaling those of vertebrates, our study of the ancient WGD event in Arachnopulmonata provides a new comparative platform to explore common and divergent evolutionary outcomes of polyploidization events across eukaryotes.

Benefits of Group Living Include Increased Feeding Efficiency and Lower Mass Loss during Desiccation in the Social and Inbreeding Spider Stegodyphus dumicola.

Group living carries a price: it inherently entails increased competition for resources and reproduction, and may also be associated with mating among relatives, which carries costs of inbreeding. Nonetheless, group living and sociality is found in many animals, and understanding the direct and indirect benefits of cooperation that override the inherent costs remains a challenge in evolutionary ecology. Individuals in groups may benefit from more efficient management of energy or water reserves, for example in the form of reduced water or heat loss from groups of animals huddling, or through reduced energy demands afforded by shared participation in tasks. We investigated the putative benefits of group living in the permanently social spider Stegodyphus dumicola by comparing the effect of group size on standard metabolic rate, lipid/protein content as a body condition measure, feeding efficiency, per capita web investment, and weight/water loss and survival during desiccation. Because energetic expenditure is temperature sensitive, some assays were performed under varying temperature conditions. We found that feeding efficiency increased with group size, and the rate of weight loss was higher in solitary individuals than in animals in groups of various sizes during desiccation. Interestingly, this was not translated into differences in survival or in standard metabolic rate. We did not detect any group size effects for other parameters, and group size effects did not co-vary with experimental temperature in a predictive manner. Both feeding efficiency and mass loss during desiccation are relevant ecological factors as the former results in lowered predator exposure time, and the latter benefits social spiders which occupy arid, hot environments.

Selection for costly sexual traits results in a vacant mating niche and male dimorphism.

The expected strong directional selection for traits that increase a male's mating ability conflicts with the frequent observation that within species, males may show extreme variation in sexual traits. These male reproductive polymorphisms are usually attributed to direct male-male competition. It is currently unclear, however, how directional selection for sexually selected traits may convert into disruptive selection, and if female preference for elaborate traits may be an alternative mechanism driving the evolution of male polymorphism. Here, we explore this mechanism using the polyandric dwarf spider Oedothorax gibbosus as a model. We first show that males characterized by conspicuous cephalic structures serving as a nuptial feeding device ("gibbosus males") significantly outperform other males in siring offspring of previously fertilized females. However, significant costs in terms of development time of gibbosus males open a mating niche for an alternative male type lacking expensive secondary sexual traits. These "tuberosus males" obtain virtually all fertilizations early in the breeding season. Individual-based simulations demonstrate a hitherto unknown general principle, by which males selected for high investment to attract females suffer constrained mating opportunities. This creates a vacant mating niche of unmated females for noninvesting males and, consequently, disruptive selection on male secondary sexual traits.

Endosymbiont dominated bacterial communities in a dwarf spider.

The microbial community of spiders is little known, with previous studies focussing primarily on the medical importance of spiders as vectors of pathogenic bacteria and on the screening of known cytoplasmic endosymbiont bacteria. These screening studies have been performed by means of specific primers that only amplify a selective set of endosymbionts, hampering the detection of unreported species in spiders. In order to have a more complete overview of the bacterial species that can be present in spiders, we applied a combination of a cloning assay, DGGE profiling and high-throughput sequencing on multiple individuals of the dwarf spider Oedothorax gibbosus. This revealed a co-infection of at least three known (Wolbachia, Rickettsia and Cardinium) and the detection of a previously unreported endosymbiont bacterium (Rhabdochlamydia) in spiders. 16S rRNA gene sequences of Rhabdochlamydia matched closely with those of Candidatus R. porcellionis, which is currently only reported as a pathogen from a woodlouse and with Candidatus R. crassificans reported from a cockroach. Remarkably, this bacterium appears to present in very high proportions in one of the two populations only, with all investigated females being infected. We also recovered Acinetobacter in high abundance in one individual. In total, more than 99% of approximately 4.5M high-throughput sequencing reads were restricted to these five bacterial species. In contrast to previously reported screening studies of terrestrial arthropods, our results suggest that the bacterial communities in this spider species are dominated by, or even restricted to endosymbiont bacteria. Given the high prevalence of endosymbiont species in spiders, this bacterial community pattern could be widespread in the Araneae order.

Spider genomes provide insight into composition and evolution of venom and silk.

Spiders are ecologically important predators with complex venom and extraordinarily tough silk that enables capture of large prey. Here we present the assembled genome of the social velvet spider and a draft assembly of the tarantula genome that represent two major taxonomic groups of spiders. The spider genomes are large with short exons and long introns, reminiscent of mammalian genomes. Phylogenetic analyses place spiders and ticks as sister groups supporting polyphyly of the Acari. Complex sets of venom and silk genes/proteins are identified. We find that venom genes evolved by sequential duplication, and that the toxic effect of venom is most likely activated by proteases present in the venom. The set of silk genes reveals a highly dynamic gene evolution, new types of silk genes and proteins, and a novel use of aciniform silk. These insights create new opportunities for pharmacological applications of venom and biomaterial applications of silk.

Spiders do not escape reproductive manipulations by Wolbachia.

BACKGROUND: Maternally inherited bacteria that reside obligatorily or facultatively in arthropods can increase their prevalence in the population by altering their hosts' reproduction. Such reproductive manipulations have been reported from the major arthropod groups such as insects (in particular hymenopterans, butterflies, dipterans and beetles), crustaceans (isopods) and mites. Despite the observation that endosymbiont bacteria are frequently encountered in spiders and that the sex ratio of particular spider species is strongly female biased, a direct relationship between bacterial infection and sex ratio variation has not yet been demonstrated for this arthropod order. RESULTS: Females of the dwarf spider Oedothorax gibbosus exhibit considerable variation in the sex ratio of their clutches and were infected with at least three different endosymbiont bacteria capable of altering host reproduction i.e. Wolbachia, Rickettsia and Cardinium. Breeding experiments show that sex ratio variation in this species is primarily maternally inherited and that removal of the bacteria by antibiotics restores an unbiased sex ratio. Moreover, clutches of females infected with Wolbachia were significantly female biased while uninfected females showed an even sex ratio. As female biased clutches were of significantly smaller size compared to non-distorted clutches, killing of male embryos appears to be the most likely manipulative effect. CONCLUSIONS: This represents to our knowledge the first direct evidence that endosymbiont bacteria, and in particular Wolbachia, might induce sex ratio variation in spiders. These findings are pivotal to further understand the diversity of reproductive phenotypes observed in this arthropod order.