Tarsal morphology and attachment ability of the codling moth Cydia pomonella L. (Lepidoptera, Tortricidae) to smooth surfaces

Despite several studies on the attachment ability of different insect taxa, little is known about this phenomenon in adult Lepidoptera. In this study we combined morphological and experimental analyses of tarsal adhesive devices and the attachment ability of the codling moth Cydia pomonella (L.) (Lepidoptera, Tortricidae) to smooth surfaces. Pretarsi of C. pomonella attach to smooth substrates by means of their smooth, flexible and well developed arolia. Using the centrifugal force measurement technique, friction forces of males and females were assessed on hydrophobic and hydrophilic glass surfaces. Adults of both sexes generated similar forces in spite of the noticeable difference in their body masses. That is why males showed significantly higher safety factors (attachment force divided by body weight) compared to those of females. Hydrophobicity of the substrate had no considerable effect on friction forces. For females, friction forces (sliding parallel to the substrate plane) were compared with adhesive forces (pulling off perpendicularly from the substrate plane) measured on Plexiglas surfaces. It can be concluded that the attachment system of C. pomonella is rather robust against physico-chemical properties of the substrate and is able to achieve a very good attachment on vertical and horizontal substrata.     

Biological microtribology: anisotropy in frictional forces of orthopteran attachment pads reflects the ultrastructure of a highly deformable material

Evolutionarily optimized frictional devices of insects are usually adapted to attach to a variety of natural surfaces. Orthopteran attachment pads are composed of hexagonal outgrowths with smooth flexible surfaces. The pads are designed to balance the weight of the insect in different positions and on different materials. In a scanning electron microscopy study followed by freezing-substitution experiments, the ultrastructural architecture of the pad material was visualized. In friction experiments, the interaction was measured between the attachment pad and a polished silicon surface. The inner structure of this material contains distally directed rods, branching close to the surface, and spaces filled with fluid. The specific design of the pad material provides a higher frictional force in the distal direction. Frictional anisotropy is more enhanced at higher normal forces and lower sliding velocities. It is concluded that optimal mechanical functionality of biosystems is the result of a combination of surface structuring and material design.     

Underwater locomotion in a terrestrial beetle: combination of surface de-wetting and capillary forces

For the first time, we report the remarkable ability of the terrestrial leaf beetle Gastrophysa viridula to walk on solid substrates under water. These beetles have adhesive setae on their feet that produce a secretory fluid having a crucial role in adhesion on land. In air, adhesion is produced by capillary forces between the fluid-covered setae and the substrate. In general, capillary forces do not contribute to adhesion under water. However, our observations showed that these beetles may use air bubbles trapped between their adhesive setae to walk on flooded, inclined substrata or even under water. Beetle adhesion to hydrophilic surfaces under water was lower than that in air, whereas adhesion to hydrophobic surfaces under water was comparable to that in air. Oil-covered hairy pads had a pinning effect, retaining the air bubbles on their feet. Bubbles in contact with the hydrophobic substrate de-wetted the substrate and produced capillary adhesion. Additional capillary forces are generated by the pad's liquid bridges between the foot and the substrate. Inspired by this idea, we designed an artificial silicone polymer structure with underwater adhesive properties.     

Attachment ability of the codling moth Cydia pomonella L. to rough substrates

Host plant surfaces of the codling moth, Cydia pomonella L. (Lepidoptera, Tortricidae), vary in microtopography, which can affect its attachment, locomotion, and oviposition behaviour. This study was performed to investigate the effect of surface roughness on the attachment ability of adult insects. Using a centrifugal force device, friction forces of both sexes were assessed on six epoxy resin substrates differing only in the dimensions of their surface asperities, ranging from 0 μm to 12 μm. Surface topography significantly affected friction forces. Maximal force was measured on the smooth substrate whereas minimal force was assessed on microrough substrates with 0.3 μm and 1.0 μm size of asperities. On the remaining rough substrates, friction forces were significantly higher but still lower than on the smooth substrate. Both sexes generated similar forces on the same substrate, in spite of the considerable difference in their body mass. Thus, it is expected that both sexes can attach effectively to differently structured plant substrates in their habitat. However, since smooth surfaces have been reported previously to be the most favorable substrates for ovipositing females of C. pomonella, it is possible that they use their attachment system to sense the substrate texture and prefer those substrates to which their arolia attach the best.     

Friction force reduction triggers feet grooming behaviour in beetles

In insects, cleaning (grooming) of tarsal attachment devices is essential for maintaining their adhesive ability, necessary for walking on a complex terrain of plant surfaces. How insects obtain information on the degree of contamination of their feet has remained, until recently, unclear. We carried out friction force measurements on walking beetles Gastrophysa viridula (Coleoptera, Chrysomelidae) and counted grooming occurrence on stiff polymer substrata with different degrees of nanoroughness (root mean square: 28-288 nm). Since nanoscopically, rough surfaces strongly reduced friction and adhesion without contaminating feet, we were able to demonstrate, for the first time to our knowledge, that friction force between tarsal attachment pads and the substrate provides an insect with information on the degree of contamination of its attachment structures. We have shown that foot grooming occurrence correlates not only with the degree of contamination but also with the decrease of friction force. This result indicates that insects obtain information about the degree of contamination, not statically but rather dynamically and, presumably, use mechanoreceptors monitoring either tensile/compressive forces in the cuticle or tensile forces between leg segments.     

Effects of surface topography and chemistry of Rumex obtusifolius leaves on the attachment of the beetle Gastrophysa viridula

Broad-leaved dock, Rumex obtusifolius L. (Polygonaceae), is the main host plant of the green dock beetle, Gastrophysa viridula Degeer (Coleoptera: Chrysomelidae). Adult beetles are able to attach and walk on leaves of this plant. Leaf surface is rather uneven, because of irregularly shaped prominent epidermal cells with a maximum height of about 9 µm. The surface is covered with a smooth epicuticular wax layer having relatively low free surface energy (FSE). The aim of this study was to measure beetle attachment force applying a 'centrifugal technique' on adult insects on the leaf surface and other substrates, in order to understand the effect of surface architecture and its physicochemical properties on insect attachment. We compared forces on an adaxial leaf surface with forces on a smooth silanized glass plate having low FSE, and on a polishing paper having slightly lower FSE and similar surface roughness (asperities' size = 9 µm). Smooth plate made of normal untreated clean glass with high FSE was used as a reference substrate. On the leaf surface and the polishing paper, attachment forces were lower compared to both glass samples. There was no significant difference between force values on leaves and the polishing paper, but on both glass surfaces they were significantly higher than on the other substrates. Hydrophobicity alone explains a decrease in attachment force of the beetle, but when combined with roughness the decrease in force is four times greater.     

Microstructure of the tarsal adhesive organs in the bark beetle Ips acuminatus,and their implication as external carriers of pathogens

Ips acuminatus is a common group of bark beetles that infest and damage pine and spruce trees. As a part of research for controlling this insect pest, the adhesive organs on the tarsal appendages were examined using field emission scanning electron microscopy (FE-SEM) to reveal the microstructural characteristics of its biological attachment system. In addition, we also demonstrate their ability to act as external carriers of pathogens. This bark beetle has a characteristic attachment apparatus to move both smooth and rough surfaces. The claws are connected with a pretarsal segment, and their apical diverged hooks are developed to hold rough substrates; however, landing on smooth surfaces is achieved by means of three groups of hairy tarsal pads. The adhesive pads are basically composed of the flattened tip setae usually with a spatula-shaped endplate. Although this bark beetle did not have mycangial cavities, yeast-like spores were concentrated at the invaginated surface of legs where cuticular hairs are densely packed. In particular, the base stalk of the adhesive pad had a sufficient space to accept spores during the dynamic movement of tenent setae.     

Effects of plant gross morphology on predator searching behaviour

Plant morphology influences insect predators' abilities to capture prey and control pest populations. Several mechanisms for this effect of plants on predator foraging have been proposed. In particular, it is often claimed that increased complexity of plant structures may increase search time and reduce foraging success. Using time-lapse photography we recorded search paths, and compared the total path lengths, percentages of plants searched, and path tortuosity of adult multicolored Asian lady beetles (Harmonia axyridis Pallas) and green lacewing larvae (Chrysoperla carnea Stephens) foraging for pea aphids (Acyrthosiphon pisum Harris) on pea near-isolines (Pisum sativum L.) that differed in shape. We found that H. axyridis searched leafy morphologies less thoroughly than those with more branches, while C. carnea larvae search paths did not differ on any of the pea morphologies. In addition, the ability of H. axyridis to attach to plants and maneuver was increased on morphologies with many branches and edges, while C. carnea was able to attach to all morphologies. Both species, however, had significantly reduced predation success on inverted leaf surfaces. We conclude that undersides of leaves, far from the leaf margin, may serve as partial prey refugia. In addition, we find increased plant branching or an increase in other morphological features which provide predator attachment points may promote foraging success.     

Scaling and biomechanics of surface attachment in climbing animals

Attachment devices are essential adaptations for climbing animals and valuable models for synthetic adhesives. A major unresolved question for both natural and bioinspired attachment systems is how attachment performance depends on size. Here, we discuss how contact geometry and mode of detachment influence the scaling of attachment forces for claws and adhesive pads, and how allometric data on biological systems can yield insights into their mechanism of attachment. Larger animals are expected to attach less well to surfaces, due to their smaller surface-to-volume ratio, and because it becomes increasingly difficult to distribute load uniformly across large contact areas. In order to compensate for this decrease of weight-specific adhesion, large animals could evolve overproportionally large pads, or adaptations that increase attachment efficiency (adhesion or friction per unit contact area). Available data suggest that attachment pad area scales close to isometry within clades, but pad efficiency in some animals increases with size so that attachment performance is approximately size-independent. The mechanisms underlying this biologically important variation in pad efficiency are still unclear. We suggest that switching between stress concentration (easy detachment) and uniform load distribution (strong attachment) via shear forces is one of the key mechanisms enabling the dynamic control of adhesion during locomotion.     

Immunocompetence of the red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae, Scolytinae): Variation between developmental stages and sexes in populations in China

Immune defense imposes fitness costs as well as benefits, so organisms should optimize, not maximize, their immune function through their life cycle. We investigated this issue in the red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae, Scolytinae), which is a pine-killing invasive beetle in China, though it is usually considered as a secondary pest in its native range of North America. We hypothesized that pathogen pressure may affect these beetles differently throughout their life history. We measured the insect's immunocompetence throughout life, determining encapsulation ability and phenoloxidase activity in larval stages, pupae and adults. Pupae had the highest encapsulation ability, but encapsulation was not different between final instar larvae and adults. Phenoloxidase (PO) activity was highest in final instar larvae and pupae, followed by the second instar larvae and adults. Total phenoloxidase activity increased significantly from the second instar larval stage to pupae, and then decreased in adults. Although the second instar larvae had the lowest phenoloxidase activity, more than 90% of total PO existed in the hemolymph in the form of the active enzyme, as compared with pupae, in which over 60% of PO occurred as a proenzyme. Both active PO and total PO were much higher in females than in males, though no significant differences were detected between the encapsulation ability of male and female adults. This result suggests the existence of a sexual dimorphism of immunocompetence in D. valens adults. Variations in immunocompetence across developmental stages suggest that D. valens adopts diverse investment strategies in immunocompetence during different stages. Potential reasons for variation in immunocompetence among developmental stages and between the sexes of D. valens are discussed.     

Fine structural analysis of the fibrillar adhesion apparatus in ladybird beetle

The attachment system on the ladybird beetle Harmonia axyridis is composed of a pair of pretarsal claws and adhesive pads at the tarsal segments. The claws, which are connected to the pretarsal segment, are mainly used to hold the rough substrates by their apical diverged hooks. In contrast, the adhesive pads have an adhesive function when landing on smooth surfaces. They are interspersed at the ventral adhesive pad of each tarsomere, and are composed of two kinds of hairy setae. The discoid tip seta (DtS) is located at the central region of each adhesive pad. The DtS has a spoon‐shaped endplate with a long and narrow shaft. In contrast, the pointed tip seta (PtS) is interspersed along the marginal regions of each adhesive pad, and has a hook‐shaped spine near the tip. In the present study, we found numerous fine cuticular pores beneath the setae, which seem to be related to the secretion of some adhesive fluids. It may be deduced that ladybird beetles can attach to smooth surfaces more effectively by employing adhesive fluids filling in surface crevices to overcome problems cause by their larger size endplates.     

Microstructure of the tarsal attachment devices in the earwig Timomenus komarovi

The biological attachment device on the tarsal appendage of the earwig, Timomenus komarovi (Insecta: Dermaptera: Forficulidae) was investigated using field emission scanning electron microscopy to reveal the fine structural characteristics of its biological attachment devices to move on smooth and rough surfaces. They attach to rough substrates using their pretarsal claws; however, attachment to smooth surfaces is achieved by means of two groups of hairy tarsal pads. This biological attachment device consists of fine hairy setae with various contact sizes. Three different groups of tenent setae were distinguished depending on the cuticular substructure of the endplates. Two groups of setae commonly had flattened surfaces, and they were covered with either spoon‐shaped or spatula‐shaped endplates, respectively. While the flattened tip setae were distributed at the central region, the pointed tip setae were characteristically found along the marginal region. There were no obvious gender‐specific differences between fibrillar adhesive pads in this insect mainly because the forceps‐like pincers are used during copulation to grasp the partner.     

Adhesive and frictional properties of tarsal attachment pads in two species of stick insects (Phasmatodea) with smooth and nubby euplantulae

In the present study, the tarsal attachment pads (euplantulae) of two stick insect species (Phasmatodea) were compared. While the euplantulae of Cuniculina impigra (syn. Medauroidea extradentata) are smooth, those of Carausius morosus bear small nubs on their surfaces. In order to characterize the adhesive and frictional properties of both types of euplantulae, adhesion and friction measurements on smooth (Ra=0.054 μm) and rough (Ra=1.399 μm) substrates were carried out. The smooth pads of C. impigra generated stronger adhesion on the smooth substrate than on the rough one. The adhesive forces of the structured pads of C. morosus did not differ between the two substrates. Friction experiments showed anisotropy for both species with higher values for proximal pulls than for distal pushes. In C. impigra, friction was stronger on the smooth than on the rough surface for both directions, whereas in C. morosus friction was stronger on the smooth surface only for pushes. This shows that smooth attachment pads are able to generate relatively stronger adhesion and friction on a flat smooth surface than on a rough one. In contrast, nubby pads have similar adhesion on both substrates, and also show no difference in friction in the pulling direction. This leads to the conclusion that smooth pads are specialized for rather smooth substrates, whereas nubby pads are better adapted to generate stronger forces on a broader range of surfaces.     

Ground reaction forces in vertically ascending beetles and corresponding activity of the claw retractor muscle on smooth and rough substrates

We measured ground reaction forces in fore–aft and normal directions of single hind and front legs in vertically ascending Sagra femorata beetles (Coleoptera, Chrysomelidae) on a smooth and a rough substrate. Simultaneously, we performed electromyographic recordings (EMGs) of the hind leg claw retractor muscle that partly controls the attachment structures. On both substrates, hind legs produced upward- as well as downward-directed forces during one stance phase. Forces were equivalent in both directions. Front legs generated only upward-directed forces. The main function of hind legs in ascending beetles in the second half of the stance thus probably prevented the animals from tilting away from the substrate. The EMGs of hind legs showed an early spike during stance with large amplitude. It was mostly followed by few additional spikes with large amplitude and in some cases of spikes with smaller amplitude distributed throughout the stance phase. We found significantly more spikes on the rough substrate than on the smooth one. This is probably due to the more important role of pretarsal claws than tarsal hairy attachment pads on the rough substrate or to the reduced adhesive forces on the rough substrate that have to be compensated by additional muscle activity.     

Two functional types of attachment pads on a single foot in the Namibia bush cricket Acanthoproctus diadematus (Orthoptera: Tettigoniidae)

Insects have developed different structures to adhere to surfaces. Most common are smooth and hairy attachment pads, while nubby pads have also been described for representatives of Mantophasmatodea, Phasmida and Plecoptera. Here we report on the unusual combination of nubby and smooth tarsal attachment structures in the !nara cricket Acanthoproctus diadematus. Their three proximal tarsal pads (euplantulae) have a nubby surface, whereas the most distal euplantula is rather smooth with a hexagonal ground pattern resembling that described for the great green bush-cricket Tettigonia viridissima. This is, to our knowledge, the first report on nubby euplantulae in Orthoptera and the co-occurrence of nubby and smooth euplantulae on a single tarsus in a polyneopteran species. When adhering upside down to a horizontal glass plate, A. diadematus attaches its nubby euplantulae less often, compared to situations in which the animal is hanging upright or head down on a vertical plate. We discuss possible reasons for this kind of clinging behaviour, such as morphological constrains, the different role of normal and shear forces in attachment enhancement of the nubby and smooth pads, ease of the detachment process, and adaptations to walking on cylindrical substrates.     

Attachment ability of sawfly larvae to smooth surfaces

Larvae of the sawfly Rhadinoceraea micans adhere properly to the anti-adhesive surface of their host plant Iris pseudacorus by using three pairs of thoracic legs, seven pairs of abdominal prolegs, and pygopodia, all provided with various smooth adhesive pads. Their attachment performance to smooth flat hydrophilic and hydrophobic glass and Plexiglas surfaces was studied in centrifugal force experiments. Obtained safety factors on Plexiglas were up to 25 in friction, and 8 in adhesion. Although larvae attached significantly stronger to the hydrophilic glass, they attached well also to the hydrophobic one. Pygopodia are suggested to dominate attachment force generation in the centrifugal force experiment. Transverse body position on the centrifuge drum was significantly advantageous for friction force generation than was longitudinal body position. Results are discussed in the context of the sawfly biology and provide a profound base for further detailed studies on biomechanics of sawfly larvae–plant interactions.     

叶甲科昆虫的定殖机制

在复杂的生态环境中,叶甲科昆虫利用寄主植物的挥发物作为嗅觉信号,并协同视觉信号共同作用以提高其远距离搜寻和定位寄主的效率。叶甲科昆虫也释放信息素来调节同种或者异种个体的行为反应;找到寄主植物后,叶子的形态学特性和化学组分的浓度等接触性因素就会影响叶甲科昆虫对寄主植物的最终选择。叶甲对上述这些信号物质的反应也受叶甲本身状态的影响,如生殖、滞育、饥饿、交配以及取食等。叶甲科昆虫对寄主植物的搜寻、辨识和接受、取食以及繁殖的过程受一种或多种因素协同影响。从嗅觉、视觉和触觉等方面对叶甲科昆虫的定殖机制做一综述。     

Host plant architecture affects the costs of dropping behaviour in <Emphasis Type="Italic">Phaedon brassicae</Emphasis> (Coleoptera: Chrysomelidae)

Phytophagous insects escape from predators by dropping. However, if they drop to the ground, they must then return to the host plants. Large oval leaves may serve as safety nets when insects drop from overlying leaves. To clarify the effects of leaf shape on the sites to which insects drop, we investigated the dropping behaviour of Phaedon brassicae (Coleoptera: Chrysomelidae) on three host plants (daikon, bok choy, and Chinese cabbage). Daikon plants have cleft leaves, whereas bok choy and Chinese cabbage plants have oval leaves. When poked with forceps, larvae dropped less frequently than adults. The proportions of individuals dropping to the ground also differed among host plants. Both larvae and adults on the ventral (abaxial) leaf surfaces of daikon frequently dropped to the ground via clefts in the underlying leaves. However, larvae and adults on the ventral leaf surfaces of bok choy and Chinese cabbage frequently dropped to underlying leaves. Most larvae and adults that dropped to the ground finally returned to host plants. However, the return times were longer for larvae than adults. Therefore, the cost of dropping from daikon leaves was higher than were the costs of dropping from leaves of other crop species.     

Activity and heavy metal resistance of non-specific esterases in leaf beetle Chrysomela lapponica from polluted and unpolluted habitats

We compared the general activity and heavy metal resistance of non-specific esterases in two populations of the leaf beetle Chrysomela lapponica from habitats severely contaminated by heavy metals (mostly Ni and Cu) and two populations from unpolluted habitats. Concentrations of Ni and Cu in adult beetles from the most polluted site were 7.7 and 3.6 times higher that in beetles from unpolluted habitats. Larval esterases showed higher activity and lower susceptibility to heavy metals than esterases of adults. Larval esterase activity did not differ between populations from polluted and unpolluted sites, but adult beetles from polluted localities had lower esterase activity than beetles from unpolluted habitats. Both Cu and Ni sulfates in millimolar concentrations in vitro suppressed esterase activity of larvae from unpolluted habitats, but caused no negative effect on esterases of larvae from polluted sites. Similarly, inhibition of adult esterase activity by Ni was stronger in beetles from unpolluted localities than in beetles from polluted localities. This indicates that resistance of non-specific esterases to heavy metals is higher in leaf beetle populations from contaminated environment.     

Preferential attachment of barnacle larvae to natural multi-species biofilms: Does surface wettability matter?

We tested the hypothesis that surface wettability does not alter the positive effect of natural biofilms on larval attachment in the barnacle Balanus amphitrite. We also answered the question: Does substratum-biofilm interaction affect the larval choice in barnacle? We developed natural multi-species biofilms of different ages on both high (glass) and low (polystyrene) wettability surfaces at mid-intertidal height (native habitat of B. amphitrite). Attachment choice of both young (0-d-old) and old (6-d-old) larvae to biofilms was determined using still water choice bioassay. Irrespective of larval age, cyprid preferred to attach to un-filmed glass than to un-filmed polystyrene. In contrast to aged larvae, young larvae preferred old (6-d-old) biofilms on polystyrene to young (3-d-old) biofilms on glass. In this study, we were also able to examine the interaction between surface wettability, biofilms and larval attachment, by characterizing bacterial community composition in biofilms. Bacterial community composition showed significant differences between biofilms of different ages. Old biofilms positively influenced larval attachment, irrespective of the type of substrata, thereby supporting the hypothesis that surface wettability does not alter the positive effect of natural biofilms on larval attachment.