Selective bactericidal activity of nanopatterned superhydrophobic cicada Psaltoda claripennis wing surfaces

The nanopattern on the surface of Clanger cicada (Psaltoda claripennis) wings represents the first example of a new class of biomaterials that can kill bacteria on contact based solely on its physical surface structure. As such, they provide a model for the development of novel functional surfaces that possess an increased resistance to bacterial contamination and infection. Their effectiveness against a wide spectrum of bacteria, however, is yet to be established. Here, the bactericidal properties of the wings were tested against several bacterial species, possessing a range of combinations of morphology and cell wall type. The tested species were primarily pathogens, and included Bacillus subtilis, Branhamella catarrhalis, Escherichia coli, Planococcus maritimus, Pseudomonas aeruginosa, Pseudomonas fluorescens, and Staphylococcus aureus. The wings were found to consistently kill Gram-negative cells (i.e., B. catarrhalis, E. coli, P. aeruginosa, and P. fluorescens), while Gram-positive cells (B. subtilis, P. maritimus, and S. aureus) remained resistant. The morphology of the cells did not appear to play any role in determining cell susceptibility. The bactericidal activity of the wing was also found to be quite efficient; 6.1?±?1.5?×?10⁶ P. aeruginosa cells in suspension were inactivated per square centimeter of wing surface after 30-min incubation. These findings demonstrate the potential for the development of selective bactericidal surfaces incorporating cicada wing nanopatterns into the design.     

Mimicking a Superhydrophobic Insect Wing by Argon and Oxygen Ion Beam Treatment on Polytetrafluoroethylene Film

Biological tiny structures have been observed on many kinds of surfaces such as lotus leaves and insect wings,whichenhance the hydrophobicity of the natural surfaces and play a role of self-cleaning.We presented the fabrication technology of asuperhydrophobic surface using high energy ion beam.Artificial insect wings that mimic the morphology and the superhydrophobocityof cicada’s wings were successfully fabricated using argon and oxygen ion beam treatment on a polytetrafluoroethylene(PTFE)film.The wing structures were supported by carbon/epoxy fibers as artificial flexible veins that were bondedthrough an autoclave process.The morphology of the fabricated surface bears a strong resemblance to the wing surface of acicada,with contact angles greater than 160°,which could be sustained for more than two months.     

Biomechanical Properties of Insect Wings: The Stress Stiffening Effects on the Asymmetric Bending of the Allomyrina dichotoma Beetle's Hind Wing

Although the asymmetry in the upward and downward bending of insect wings is well known, the structural origin of this asymmetry is not yet clearly understood. Some researchers have suggested that based on experimental results, the bending asymmetry of insect wings appears to be a consequence of the camber inherent in the wings. Although an experimental approach can reveal this phenomenon, another method is required to reveal the underlying theory behind the experimental results. The finite element method (FEM) is a powerful tool for evaluating experimental measurements and is useful for studying the bending asymmetry of insect wings. Therefore, in this study, the asymmetric bending of the Allomyrina dichotoma beetle''s hind wing was investigated through FEM analyses rather than through an experimental approach. The results demonstrated that both the stressed stiffening of the membrane and the camber of the wing affect the bending asymmetry of insect wings. In particular, the chordwise camber increased the rigidity of the wing when a load was applied to the ventral side, while the spanwise camber increased the rigidity of the wing when a load was applied to the dorsal side. These results provide an appropriate explanation of the mechanical behavior of cambered insect wings, including the bending asymmetry behavior, and suggest an appropriate approach for analyzing the structural behavior of insect wings.     

Mechanism of the wing colouration in the dragonfly Zenithoptera lanei (Odonata: Libellulidae) and its role in intraspecific communication

Zenithoptera dragonflies are known for their remarkable bluish colouration on their wings and unique male behaviour of folding and unfolding their wings while perching. However, nothing is known about the optical properties of such colouration and its structural and functional background. In this paper, we aimed to study the relationship between the wing membrane ultrastructure, surface microstructure and colour spectra of male wings in Zenithoptera lanei and test the hypothesis that colouration functions as a signal in territorial fights between males. The results show that the specific wing colouration derives from interference in alternating layers of melanized and unmelanized cuticle in the wing membrane, combined with diffuse scattering in two different layers of wax crystals on the dorsal wing surface, one lower layer of long filaments, and one upper layer of leaf-shaped crystals. The results also show that the thicker wax coverage of the dorsal surface of the wings results in increased brightness and reduced chroma. In the field experiments, we have demonstrated that there is a reduction of aggressive reactions of rivals towards individuals with experimentally reduced amount of blue wing colouration.     

Supernumerary limb structures after juxtaposing dorsal and ventral chick wing bud cells

The formation of duplicated wing skeletal elements and/or extra wing muscles was studied by juxtaposing normally nonadjacent embryonic chick wing bud cells. A wedge of right or left stage 21 wing bud ectoderm and mesoderm was inserted in a slit made in a host stage 20 to 22 right wing bud at the same anteroposterior position as its position of origin. The distal edge of the donor wedge and host wing bud were aligned with each other. Donor tissue was grafted into a host wing bud in one of the following four axial relationships: both the anteroposterior and dorsoventral axes corresponded with each other (aadd); only the anteroposterior axes were opposed (apdd); only the dorsoventral axes were opposed (aadv); both the anteroposterior and dorsoventral axes were opposed (apdv). Of the 63 wings resulting from the control aadd operation and the 45 wings from the apdd operation, only 12 wings had a duplicated skeletal element; of the 69 wings sectioned from these two groups of operations, only one had an extra muscle. However, of the wings resulting from the aadv and apdv operations (48 and 52 cases, respectively), 23 had a duplicated skeletal element; of the 54 wings sectioned from these operations, 43 wings had one to four extra muscles. Furthermore, when the aadv operation was performed with a wedge of donor quail wing bud ectoderm and mesoderm or mesoderm alone, supernumerary muscles formed in these chimeric wings and they were made up of donor quail and host chick cells or only donor quail cells.     

Prevalence of Campylobacter jejuni in Chicken Wings

Campylobacter jejuni was found in 82.9% of 94 chicken wing packages analyzed on the day of arrival at supermarkets and in 15.5% of 45 packages obtained from the supermarket shelves a few days later. The number of bacterial cells ranged from 10² to 10^3.9 per wing. The prevalence of C. jejuni in the wings varied with the brand, the day of sampling, and the age of the product.     

Structural Characteristics of Allomyrina Dichotoma Beetle＇s Hind Wings for Flapping Wing Micro Air Vehicle

In this study, we present a complete structural analysis ofAllomyrina dichotoma beetle＇s hind wings by investigating their static and dynamic characteristics. The wing was subjected to the static loading to determine its overall flexural stiffness. Dy- namic characteristics such as natural frequency, mode shape, and damping ratio of vibration modes in the operating frequency range were determined using a Bruel ＆ Kjaer fast Fourier transform analyzer along with a laser sensor. The static and dynamic characteristics of natural Allomyrina dichotoma beetle＇s hind wings were compared to those of a fabricated artificial wing. The results indicate that natural frequencies of the natural wing were significantly correlated to the wing surface area density that was defined as the wing mass divided by the hind wing surface area. Moreover, the bending behaviors of the natural wing and artificial wing were similar to that of a cantilever beam. Furthermore, the flexural stiffness of the artificial wing was a little higher than that of the natural one whereas the natural frequency of the natural wing was close to that of the artificial wing. These results provide important information for the biomimetic design of insect-scale artificial wings, with which highly ma- neuverable and efficient micro air vehicles can be designed.     

Putative functions and functional efficiency of ordered cuticular nanoarrays on insect wings

The putative functions and functional efficiencies of periodic nanostructures on the surface of cicada wings have been investigated by atomic force microscopy (AFM) used as a tool for imaging, manipulation, and probing of adhesion. The structures consist of hexagonal close-packed protrusions with a lateral spacing of ∼200 nm and may have multiple functionalities. Not only do the structures confer survival value by virtue of camouflage, but they may also serve as antiwetting and self-cleaning surfaces and thus be resistant to contamination. These effects have been demonstrated by exposure to white light, liquid droplets, and AFM adhesion measurements. The dependence of optical reflectivity and surface adhesion on surface topography has been demonstrated using AFM as a nanomachining tool as well as an imaging and force-sensing probe. The intact arrays display exceptionally low adhesion for particles in the size range 20 nm-40 μm. The particles can be removed from the array by forces in the range 2-20 nN; conversely, forces in the range 25-230 nN are required to remove identical particles from a flat hydrophilic surface (i.e., polished Si). Measurements of contact angles for several liquids and particle adhesion studies show that the wing represents a low-surface-energy membrane with antiwetting properties. The inference is that a combination of chemistry and structure constitutes a natural technology for conferring resistance to contamination.     

Studies on the female-sterile mutant rudimentary of Drosophila melanogaster,: I. An analysis of the rudimentary wing phenotype

The rudimentary wing phenotype was examined in detail, using six different alleles of rudimentary, and a number of points about the genesis of the r phenotype were made. (1) All of the r alleles in which the wings are defective produce wings in which the area of individual hair cells is reduced. The more severely affected the allele, the greater is the reduction in wing cell area. This reduction in area is probably uniform throughout the wing rather than localized to specific wing regions. (2) The total number of cells per wing is also greatly reduced in phenotypically r wings. As with cell area, the more severely affected the allele, the greater the reduction in cell number. However, the reduction in cell number is not uniform throughout the wing. In the less severely affected alleles, the cell number reduction is much greater in those regions of the wing which are drastically altered in shape (truncated), while those wing regions which show only slight size reductions but no overall shape changes have near normal numbers of cells. In the most deformed wings, there is a reduction in cell number throughout the wing, but again those regions with are severely truncated are the most drastically reduced in cell number. Measurements of the amount of chitin per wing indicated that the three most severely affected alleles had as much or more chitin than the wild type. It is suggested that overproduction of chitin in these alleles prevents normal expansion of the wing cells, thus increasing the severity of the wing defect. Finally, the validity and limitations of a quantitative measure of the r phenotype were defined. This measure was utilized to demonstrate a clear-cut effect of nutrition on the expression of the r phenotype.     

A study of the anti-reflection efficiency of natural nano-arrays of varying sizes

The dependence of optical reflectivity and wettability on the surface topography of 32 species of cicada wing membranes has been investigated using UV-visible spectrophotometry, contact angle measurements and environmental scanning electron microscopy. The nanoscale hexagonally close packed protrusions have been shown to exhibit an anti-reflection and in some cases an anti-wetting function. The parameters of the structures were measured to be 77-148 nm in diameter, 44-117 nm in spacing and 159-481 nm in height. The transmittance spectrum and static contact angles were measured. At a wavelength range of 500-2500 nm, only minor differences in the anti-reflection performance were observed for each cicada species ascribed to the mechanism of impedance matching between cuticle and air. The transmittance properties of cicada wings were altered successfully through the scanning probe microscope-based manipulation by reducing the protrusion height via the contact mode. A near linear dependence was found between a decrease in protuberance height and a resulting increase in reflectance intensity. A diversity of wettability was observed with contact angles varying from 56.5° to 146.0°. Both effects of anti-reflection and wettability are dependent on the height of protrusions. The anti-reflection is insensitive when the wavelength is larger than the lateral feature size of the nanostructure. The stronger hydrophobic properties are generally associated with a larger diameter, closer spacing and greater height of protrusions when the wing membrane is intact.     

A nondestructive method of calculating the wing area of insects

Most insects engage in winged flight. Wing loading, that is, the ratio of body mass to total wing area, has been demonstrated to reflect flight maneuverability. High maneuverability is an important survival trait, allowing insects to escape natural enemies and to compete for mates. In some ecological field experiments, there is a need to calculate the wing area of insects without killing them. However, fast, nondestructive estimation of wing area for insects is not available based on past work. The Montgomery equation (ME), which assumes a proportional relationship between leaf area and the product of leaf length and width, is frequently used to calculate leaf area of plants, in crops with entire linear, lanceolate leaves. Recently, the ME was proved to apply to leaves with more complex shapes from plants that do not have any needle leaves. Given that the wings of insects are similar in shape to broad leaves, we tested the validity of the ME approach in calculating the wing area of insects using three species of cicadas common in eastern China. We compared the actual area of the cicadas’ wings with the estimates provided by six potential models used for wing area calculation, and we found that the ME performed best, based on the trade‐off between model structure and goodness of fit. At the species level, the estimates for the proportionality coefficients of ME for three cicada species were 0.686, 0.693, and 0.715, respectively. There was a significant difference in the proportionality coefficients between any two species. Our method provides a simple and powerful approach for the nondestructive estimation of insect wing area, which is also valuable in quantifying wing morphological features of insects. The present study provides a nondestructive approach to estimating the wing area of insects, allowing them to be used in mark and recapture experiments.     

Morphometrics of the final instar exuviae of five cicada species occurring in urban areas of central Korea

Cicadas produce loud calling songs for mate attraction that can be a nuisance to city dwellers in Korea. Exuviae of final instars can be used to estimate population density. We investigated the morphological characteristics of final instar exuviae for five cicada species that are abundant in central Korea: Cryptotympana atrata, Hyalessa fuscata, Graptopsaltria nigrofuscata, Meimuna opalifera, and Meimuna mongolica. The characters analyzed were pro-mesonotum length, pronotum width, pronotum length, head width, abdominal circumference, body length, wing length, and femoral tooth angle. The results of a general linear model showed that species and sex were significant for morphological characters, and the result of post hoc analyses revealed that the mean values of most of the morphological characters measured were significantly different among the five cicada species. Three groups, Cryptotympana atrata, H. fuscata/G. nigrofuscata, and M. opalifera/M. mongolica, were distinctively identified based on most morphological characters. The distributions of femoral tooth angles did not overlap between H. fuscata and G. nigrofuscata, and the distributions of abdominal circumference did not overlap between M. opalifera and M. mongolica. Thus, the exuviae of all five cicada species could be easily distinguished based on morphological characters. We also provide a morphological key for the exuviae of all five cicada species in Korea.     

The Drosophila wing hearts originate from pericardial cells and are essential for wing maturation

In addition to the heart proper, insects possess wing hearts in the thorax to ensure regular hemolymph flow through the narrow wings. In Drosophila, the wing hearts consist of two bilateral muscular pumps of unknown origin. Here, we present the first developmental study on these organs and report that the wing hearts originate from eight embryonic progenitor cells arising in two pairs in parasegments 4 and 5. These progenitors represent a so far undescribed subset of the Even-skipped positive pericardial cells (EPC) and are characterized by the early loss of tinman expression in contrast to the continuously Tinman positive classical EPCs. Ectopic expression of Tinman in the wing heart progenitors omits organ formation, indicating a crucial role for Tinman during progenitor specification. The subsequent postembryonic development is a highly dynamic process, which includes proliferation and two relocation events. Adults lacking wing hearts display a severe wing phenotype and are unable to fly. The phenotype is caused by omitted clearance of the epidermal cells from the wings during maturation, which inhibits the formation of a flexible wing blade. This indicates that wing hearts are required for proper wing morphogenesis and functionality.     

Morphological integration and modularity in Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) hind wings

The morphological integration of the hind wings of the western corn rootworm Diabrotica virgifera virgifera LeConte was investigated to get a better insight of the undergone by this invasive species. Geometric morphometric methods were used to test two modularity hypotheses associated with the wing development and function (hypothesis H1: anterior/posterior or H2: distal/proximal wing parts). Both hypotheses were rejected and the results showed the integrated behavior of the hind wings of D. v. virgifera. The hypothesized modules do not represent separate units of variation, so in a similar fashion as exhibited by the model species Drosophila melanogaster, the hind wings of D. v. virgifera act as a single functional unit. The moderate covariation strength found between anterior and posterior and distal and proximal parts of the hind wing of D. v. virgifera confirms its integrated behavior. We conclude that the wing shape shows internal integration, which could enable flexibility and thus enhance flight maneuverability. This study contributes to the understanding of morphological integration and modularity on a non-model organism. Additionally, these findings lay the groundwork for future flight performance and biogeographical studies on how wing shape and size vary across the endemic and expanded/invaded range in the USA and Europe infested with D. v. virgifera.     

Lack of response to artificial selection on developmental stability of partial wing shape components in Drosophila melanogaster

Developmental stability, the ability of organisms to buffer their developmental processes against developmental noise is often evaluated with fluctuating asymmetry (FA). Natural genetic variation in FA has been investigated using Drosophila wings as a model system and the recent estimation of the heritability of wing shape FA was as large as 20 %. Because natural genetic variation in wing shape FA was found to localize in a partial component of the wings, heritable variation in specific parts of the wings might be responsible for FA estimation based on the whole wing shape. In this study, we quantified the shape of three partial components of the wings, and estimated the heritability of the wing shape FA based on artificial selections. As a result, FA values for the partial wing shape components did not respond to artificial selections and the heritability scores estimated were very small. These results indicate that natural additive genetic variation in FA of partial wing components was very small compared with that in a complex wing trait.     

Sexual Dichromatism of the Damselfly Calopteryx japonica Caused by a Melanin-Chitin Multilayer in the Male Wing Veins

Mature male Calopteryx japonica damselflies have dark-blue wings, due to darkly coloured wing membranes and blue reflecting veins. The membranes contain a high melanin concentration and the veins have a multilayer of melanin and chitin. Female and immature C. japonica damselflies have brown wings. We have determined the refractive index of melanin by comparing the differently pigmented wing membranes and applying Jamin-Lebedeff interference microscopy. Together with the previously measured refractive index of chitin the blue, structural colour of the male wing veins could be quantitatively explained by an optical multilayer model. The obtained melanin refractive index data will be useful in optical studies on melanized tissues, especially where melanin is concentrated in layers, thus causing iridescence.     

Kinematic compensation for wing loss in flying damselflies

Flying insects can tolerate substantial wing wear before their ability to fly is entirely compromised. In order to keep flying with damaged wings, the entire flight apparatus needs to adjust its action to compensate for the reduced aerodynamic force and to balance the asymmetries in area and shape of the damaged wings. While several studies have shown that damaged wings change their flapping kinematics in response to partial loss of wing area, it is unclear how, in insects with four separate wings, the remaining three wings compensate for the loss of a fourth wing. We used high-speed video of flying blue-tailed damselflies (Ischnura elegans) to identify the wingbeat kinematics of the two wing pairs and compared it to the flapping kinematics after one of the hindwings was artificially removed. The insects remained capable of flying and precise maneuvering using only three wings. To compensate for the reduction in lift, they increased flapping frequency by 18 ± 15.4% on average. To achieve steady straight flight, the remaining intact hindwing reduced its flapping amplitude while the forewings changed their stroke plane angle so that the forewing of the manipulated side flapped at a shallower stroke plane angle. In addition, the angular position of the stroke reversal points became asymmetrical. When the wingbeat amplitude and frequency of the three wings were used as input in a simple aerodynamic model, the estimation of total aerodynamic force was not significantly different (paired t-test, p = 0.73) from the force produced by the four wings during normal flight. Thus, the removal of one wing resulted in adjustments of the motions of the remaining three wings, exemplifying the precision and plasticity of coordination between the operational wings. Such coordination is vital for precise maneuvering during normal flight but it also provides the means to maintain flight when some of the wings are severely damaged.     

Nanomechanical properties of wing membrane layers in the house cricket (Acheta domesticus Linnaeus)

Many insect wings change shape dynamically during the wingbeat cycle, and these deformations have the potential to confer energetic and aerodynamic benefits during flight. Due to the lack of musculature within the wing itself, the changing form of the wing is determined primarily by its passive response to inertial and aerodynamic forces. This response is in part controlled by the wing’s mechanical properties, which vary across the membrane to produce regions of differing stiffness. Previous studies of wing mechanical properties have largely focused on surface or bulk measurements, but this ignores the layered nature of the wing. In our work, we investigated the mechanical properties of the wings of the house cricket (Acheta domesticus) with the aim of determining differences between layers within the wing. Nanoindentation was performed on both the surface and the interior layers of cross-sectioned samples of the wing to measure the Young’s modulus and hardness of the outer- and innermost layers. The results demonstrate that the interior of the wing is stiffer than the surface, and both properties vary across the wing.     

Does Skipping a Meal Matter to a Butterfly's Appearance? Effects of Larval Food Stress on Wing Morphology and Color in Monarch Butterflies

In animals with complex life cycles, all resources needed to form adult tissues are procured at the larval stage. For butterflies, the proper development of wings involves synthesizing tissue during metamorphosis based on the raw materials obtained by larvae. Similarly, manufacture of pigment for wing scales also requires resources acquired by larvae. We conducted an experiment to test the effects of food deprivation in the larval stage on multiple measures of adult wing morphology and coloration of monarch butterflies (Danaus plexippus), a species in which long-distance migration makes flight efficiency critical. In a captive setting, we restricted food (milkweed) from late-stage larvae for either 24 hrs or 48 hrs, then after metamorphosis we used image analysis methods to measure forewing surface area and elongation (length/width), which are both important for migration. We also measured the brightness of orange pigment and the intensity of black on the wing. There were correlations between several wing features, including an unexpected association between wing elongation and melanism, which will require further study to fully understand. The clearest effect of food restriction was a reduction in adult wing size in the high stress group (by approximately 2%). Patterns observed for other wing traits were ambiguous: monarchs in the low stress group (but not the high) had less elongated and paler orange pigmentation. There was no effect on wing melanism. Although some patterns obtained in this study were unclear, our results concerning wing size have direct bearing on the monarch migration. We show that if milkweed is limited for monarch larvae, their wings become stunted, which could ultimately result in lower migration success.     

Restoration of wing margins in Lyra mutants of Drosophila melanogaster

Lyra is a dominant, homozygous lethal mutation of Drosophila melanogaster; in heterozygotes the wings lack portions of the anterior and posterior margins including the characteristic bristles. We have found that, in addition to the loss of bristle forming cells, there is a decrease in the number of wing surface cells that varies between 10 and 20%. However, we observed no histological evidence of excessive cell death in either the larval discs or the pupal wing precursors in Lyra flies. Restoration of all or part of the normal wing margins occurs in some, but not all, cases of morphogenetic mosaics, in which there were patches of wild-type cells in Lyra wing margins due to irradiation-induced mitotic recombination. Analysis of these restorations, using margin bristles as indicators, shows that the Lyra wild-type gene is not involved in bristle formation per se and further that its expression is not cell autonomous. Instead the effect of the Lyra mutation appears to be associated with development of a margin forming subpopulation of cells and to influence the characteristic pattern of cells and bristles in the wing margin via an inductive interaction. The dorsal-ventral boundary can be demonstrated in the de facto wing margins of Lyra mutants suggesting that its origin is independent of any function Lyra might have in normal wing margin morphogenesis. In wing margin restorations the dorsal-ventral boundary is clearly delimited by trichomes and somewhat less rigorously shown by the margin bristles. Further, in these restorations ventral clones induce dorsal bristles, as well as ventral ones, and vice versa, indicating that the influence of Lyra is not restricted by the dorsal-ventral boundary.