Effect of sample treatment on the elastic modulus of locust cuticle obtained by nanoindentation

Cuticle is one of the most abundant, but least studied, biological composites. As a result, it has contributed very little to the field of biomimetics. An important step to overcome this problem is to study cuticle biomechanics by means of accurate mechanical measurements. However, due to many reasons, mechanical testing on fresh cuticle specimens is not always possible. Hence, researchers often use stored specimens to measure properties of arthropod cuticle. Our knowledge about the influence of different treatment methods on cuticle properties is currently very limited. In this study, we investigated the effect of freezing, desiccation, and rehydration on the elastic modulus of the hind tibial cuticle of locusts obtained by nanoindentation. We found that all the mentioned treatments significantly influence cuticle properties. This is in contrast to previous reports suggesting that freezing did not significantly influence the elastic modulus of native cuticle specimens tested in bending. In the light of our data, we suggest that changes of the elastic modulus of cuticle are not solely due to changes of the water content. Our results provide a platform for more accurate measurements of cuticle properties

Conflicting requirements for transparency and mechanical stability in the compound eyes of desert locusts

Compound eyes of insects should be both thin and transparent to allow light to pass through, and at the same time mechanically stable to serve as exoskeleton. These conflicting requirements make the corneal cuticle an interesting example for studying cuticle biomechanics as well as for designing composite materials that seek similar properties. Here we combined scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and nanoindentation, to investigate the microstructure, material composition and material properties of the corneal cuticle of desert locust Schistocerca gregaria. The results suggest that a fully helicoidal architecture and large proportion of resilin in the corneal cuticle are likely to be adaptations for light transmission. Even though the corneal cuticle is resilin-rich, its elastic modulus is at least three times higher than that previously reported for other resilin-rich cuticles. This is likely due to the specific microstructure of the corneal cuticle with densely packed layers. This study presents one of a series of studies, in which we used multidisciplinary approaches, to understand the link between the structure, material, property, and function in insect cuticle

WingSegment: A Computer Vision‐Based Hybrid Approach for Insect Wing Image Segmentation and 3D Printing

This article introduces WingSegment, a MATLAB app‐designed tool employing a hybrid approach of computer vision and graph theory for precise insect wing image segmentation. WingSegment detects cells, junctions, Pterostigma, and venation patterns, measuring geometric features and generating Voronoi patterns. The tool utilizes region‐growing, thinning, and Dijkstra's algorithms for boundary detection, junction identification, and vein path extraction. It provides histograms and box plots of geometric features, facilitating comprehensive wing analysis. WingSegment's efficiency is validated through comparisons with established tools and manual measurements, demonstrating accurate results. The tool further enables exporting detected boundaries as FreeCAD macro files for 3D modeling and printing, supporting finite element analysis. Beyond advancing insect wing morphology understanding, WingSegment holds broader implications for diverse planar structures, including leaves and geocells. This tool not only enhances automated geometric analysis and 3D model generation in insect wing studies but also contributes to the broader advancement of analysis, 3D printing, and modeling technologies across various planar structures

Reduction of female copulatory damage by resilin represents evidence for tolerance in sexual conflict

Intergenomic evolutionary conflicts increase biological diversity. In sexual conflict, female defence against males is generally assumed to be resistance, which, however, often leads to trait exaggeration but not diversification. Here, we address whether tolerance, a female defence mechanism known from interspecific conflicts, exists in sexual conflict. We examined the traumatic insemination of female bed bugs via cuticle penetration by males, a textbook example of sexual conflict. Confocal laser scanning microscopy revealed large proportions of the soft and elastic protein resilin in the cuticle of the spermalege, the female defence organ. Reduced tissue damage and haemolymph loss were identified as adaptive female benefits from resilin. These did not arise from resistance because microindentation showed that the penetration force necessary to breach the cuticle was significantly lower at the resilin-rich spermalege than at other cuticle sites. Furthermore, a male survival analysis indicated that the spermalege did not impose antagonistic selection on males. Our findings suggest that the specific spermalege material composition evolved to tolerate the traumatic cuticle penetration. They demonstrate the importance of tolerance in sexual conflict and genitalia evolution, extend fundamental coevolution and speciation models and contribute to explaining the evolution of complexity. We propose that tolerance can drive trait diversity

Patterns of load distribution among the legs in small water striders during standing and striding

Water striders (Gerris argentatus) move across the water surface by taking advantage of the surface tension, which supports their bodyweight without breaking. During locomotion, the midlegs are primarily responsible for generating thrust, whereas the other legs support the body. Although the aspects of standing and locomotion on the water surface are well understood, relatively fewer studies concerned the coordinated biomechanical movements of the legs. In order to maintain buoyancy of the body on the water surface, the leg positions must be adjusted to distribute the bodyweight appropriately. The present study investigates distribution of the bodyweight on the legs in relatively small water striders. We aimed to understand how loading on the legs changes during sculling that leads to sliding of the body on the water surface. The assistance of all legs at every moment enables the body to maintain its floating during standing and striding. Water striders can achieve a gentle striding through the midlegs driving phase in association with smooth load shifting among their legs, which are positioned in a specific configuration to support the insect on the water surface

Extensive collection of femtoliter pad secretion droplets in beetle Leptinotarsa decemlineata allows nanoliter microrheology

Pads of beetles are covered with long, deformable setae, each ending in a micrometric terminal plate coated with secretory fluid. It was recently shown that the layer of the pad secretion covering the terminal plates is responsible for the generation of strong attractive forces. However, less is known about the fluid itself because it is produced in extremely small quantity. We here present a first experimental investigation of the rheological properties of the pad secretion in the Colorado potato beetle {\it Leptinotarsa decemlineata}. Because the secretion is produced in an extremely small amount at the level of the terminal plate, we first develop a procedure based on capillary effects to collect the secretion. We then manage to incorporate micrometric beads, initially in the form of a dry powder, and record their thermal motion to determine the mechanical properties of the surrounding medium. We achieve such a quantitative measurement within the collected volume, much smaller than the $1 {\rm \mu}$l sample volume usually required for this technique. Surprisingly, the beetle secretion was found to behave as a purely viscous liquid, of high viscosity. This suggests that no specific complex fluid behaviour is needed during beetle locomotion. We build a scenario for the contact formation between the spatula at the setal tip and a substrate, during the insect walk. We show that the attachment dynamics of the insect pad computed from the high measured viscosity is in good agreement with observed insect pace. We finally discuss the consequences of the secretion viscosity on the insect adhesion

Locomotory Behavior of Water Striders with Amputated Legs

The stability of the body during locomotion is a fundamental requirement for walking animals. The mechanisms that coordinate leg movement patterns are even more complex at water–air interfaces. Water striders are agile creatures on the water surface, but they can be vulnerable to leg damage, which can impair their movement. One can assume the presence of certain compensatory biomechanical factors that are involved in the maintenance of postural balance lost after an amputation. Here, we studied changes in load distribution among the legs and assessed the effects of amputation on the locomotory behavior and postural defects that may increase the risk of locomotion failure. Apparently, amputees recover a stable posture by applying leg position modifications (e.g., widening the stance) and by load redistribution to the remaining legs. Water striders showed steering failure after amputation in all cases. Amputations affected locomotion by (1) altering motion features (e.g., shorter swing duration of midlegs), (2) functional constraints on legs, (3) shorter travelled distances, and (4) stronger deviations in the locomotion path. The legs functionally interact with each other, and removal of one leg has detrimental effects on the others. This research may assist the bioinspired design of aquatic robots

The Influence of Material and Roughness on the Settlement and the Adhesive Strength of the Barnacle Balanus Improvisus in the Baltic Sea

Under natural conditions, barnacles, one of the most prominent marine hardfouling organisms, encounter a vast variety of heterogeneous surfaces including artificial ones, such as ship hulls on which they adhere efficiently. Despite intensive research in the last decades, it is still not clear, how material related factors influence the fouling development under natural conditions, and whether roughness at the micro scale affects the release dynamics of barnacles. In order to shed light on the relationship between these substrate factors and their effect on biofouling, both settlement and fouling development of Balanus (= Amphibalanus) improvisus was evaluated on epoxy resin and polyvinylsiloxane (PVS) substrates differing in their roughness (flat, asperity sizes: 0.3, 1, 3, 9, 12 μm) in a static field trial in the Baltic Sea for 17 weeks performed in 2014. All barnacles on these tested surfaces were individually tracked on a weekly basis, in order to calculate the release-to-settlement ratios (r/s), to evaluate the fouling- release performance and to monitor the actual attachment duration. It was demonstrated that both stiffness and surface free energy had no strong effect on initial settlement. The total fouling accumulation was nearly identical for epoxy resin and PVS, if fallen-off barnacles were included in the analysis. Roughness influenced initial settlement and fouling development. On PVS the r/s ranged between 0.5 and 0.7 and the attachment duration was around 4 weeks. However, samples with a roughness of 9 μm displayed a lower r/s ratio (0.35) and barnacles showed longer attachment durations (8 weeks). In a second field trial performed in 2017, attachment forces of barnacles were measured after the samples had been immersed for 10 weeks in the Baltic Sea. The shear stresses obtained were similar for substrates with different roughness with around 0.12 MPa and were independent of the barnacle’s size. These findings show that roughness even at the micro scale can impact the fouling release ability of a surface

The frequency of wing damage in a migrating butterfly

The ability to fly is crucial for migratory insects. Consequently, the accumulation of damage on the wings over time can affect survival, especially for species that travel long distances. We examined the frequency of irreversible wing damage in the migratory butterfly Vanessa cardui to explore the effect of wing structure on wing damage frequency, as well as the mechanisms that might mitigate wing damage. An exceptionally high migration rate driven by high precipitation levels in their larval habitats in the winter of 2018–2019 provided us with an excellent opportunity to collect data on the frequency of naturally occurring wing damage associated with long-distance flights. Digital images of 135 individuals of V. cardui were collected and analyzed in Germany. The results show that the hindwings experienced a greater frequency of damage than the forewings. Moreover, forewings experienced more severe damage on the lateral margin, whereas hindwings experienced more damage on the trailing margin. The frequency of wing margin damage was higher in the painted lady butterfly than in the migrating monarch butterfly and in the butterfly Pontia occidentalis following artificially induced wing collisions. The results of this study could be used in future comparative studies of patterns of wing damage in butterflies and other insects. Additional studies are needed to clarify whether the strategies for coping with wing damage differ between migratory and non-migratory species